AIM:To evaluate effects of endogenous dopamine induced by low concentration atropine eye drops on choroidal neovascularization(CNV)in high myopia mice.METHODS:The C57BL/6J mice were deprived of the right eye for 4wk,a...AIM:To evaluate effects of endogenous dopamine induced by low concentration atropine eye drops on choroidal neovascularization(CNV)in high myopia mice.METHODS:The C57BL/6J mice were deprived of the right eye for 4wk,and the high myopia was diagnosed by optometry,the diopter was less than-6.00 D,and CNV was induced by 532 nm laser.The changes of dopamine D1 receptor(DRD1),dopamine D2 receptor(DRD2),and vascular endothelial growth factor A(VEGFA)were detected by Western blot technology at 0.5,1,2h,and 7d after 0.01%,0.05%,and 0.1%atropine eye drops,respectively,the area of CNV was measured.RESULTS:Significant increases were observed on the expression of DRD2 in mouse high myopia model at 0.5,1,2h,7d with 0.05%and 0.1%atropine eye drops(P<0.05).Significant decreases were observed on the expression of DRD1 and VEGFA in mouse high myopia model at 0.5,1,2h,7d with 0.05%and 0.1%atropine eye drops(P<0.05).The area of CNV induced by laser in the drug-treated group was significantly smaller than that in the control group,and the higher the concentration,the more significant the inhibitory effect(P<0.05).CONCLUSION:The 0.01%,0.05%,0.1%atropine eye drops can decrease the level of VEGFA and inhibit high myopia CNV indirectly by up-regulating the level of DRD2 and down-regulating the level of DRD1,and the effect of 0.05%and 0.1%atropine eye drops is more significant.展开更多
AIM:To evaluate the efficacy of intravitreal injection of anti-vascular endothelial growth factor(anti-VEGF),photodynamic therapy(PDT),and laser treatment(LT)for anatomical and functional improvement in myopic choroid...AIM:To evaluate the efficacy of intravitreal injection of anti-vascular endothelial growth factor(anti-VEGF),photodynamic therapy(PDT),and laser treatment(LT)for anatomical and functional improvement in myopic choroidal neovascularization(mCNV)patients.METHODS:Two researchers independently searched PubMed,Cochrane Library,Web of Science,and other databases to screen studies comparing best-corrected vision acuity(BCVA)and foveal center thickness(FCT)changes after mCNV treatment.Post-treatment chorioretinal atrophy(CRA)is a secondary outcome indicator.The retrieval time limit is from the database construction to January 30,2023.RESULTS:A total of 1072 eyes in 16 articles were included.In the RCTs,intravitreal bevacizumab(IVB)and intravitreal ranibizumab(IVR)were superior to PDT(MD=0.18,95%CI:0.02,0.40,MD=0.18,95%CI:0.01,0.42)in improving BCVA of mCNV patients(P<0.05).The relative effectiveness in improving BCVA,from high to low,appeared to be IVR,intravitreal aflibercept(IVA),IVB,LT,PDT,and sham first followed by IVA(Sham/IVA).While improving the FCT from high to low was IVA,IVR,IVB,PDT.In retrospective studies,the results of BCVA after long-term treatment showed that all the therapeutic effects from high to low was IVA,intravitreal conbercept(IVC),IVR,IVB,IVB/IVR,PDT with IVB/IVR,PDT.The effect of improving FCT was IVA,IVR,IVC,PDT,and IVB from high to low.And in the effects of improving CRA,the IVB appeared to be higher than IVR,while the PDT was the smallest,but none of the differences in the results were statistically significant.CONCLUSION:Anti-VEGF has the best effect on longterm vision improvement in mCNV patients,using IVB or IVR alone to treat mCNV may be better than IVB or IVR combined with PDT.There is no significant difference in the improvement of visual acuity,macular edema,and CRA in mCNV patients treated with any different anti-VEGF drugs.展开更多
AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue te...AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue tetrazolium bromide(M一)assay.Effects of RSV on choroidal endothelial cell(CEC)migration were evaluated with a modified Boyden chamber assay,while tube formation was evaluated in a 2-D gel assay.CNV was induced by laser photocoagulation in mice.The effects of intravitreal injection of RSV on CNV development were evaluated by fluorescein angiography(FA),confocal analysis of isolectin B4 labeled choroidal flat mounts,and histologic examination of CNV membranes.Immunostaining was used to analyze the expression and phosphorylation of vascular endothelial growth factor receptor 2(VEGFR2).RESULTS:No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 pmol/L(P>0.05).RSV inhibited vascular endothelial growth factor(VEGF)-induced CEC migration(P<0.05)and tube formation(P<0.05)invitro.Furthermore,intravitrealinjectionof RSV significantly inhibited laser induced CNV formation in mice.The FA leakage,CNV volume and CNV area analysis revealed that there were 41%,45%,and 58%reduction in RSV-treated eyes(1.691±0.1032,178163±78623μm^3 and 6508±619.0μm^2,respectively)compared with those in control(2.724±0.08447,379676±98382μm3and16576±2646μm^2,respectively;P<0.05).Phospho-VEGFR2expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control(P<0.05).CONCLUSION:Intravitreal injection of RSV exerts an inhibitory effect on CNV,which may through suppressing endothelial cell migration,tube formation and VEGFR2 phosphorylation.展开更多
Purpose: Choroidal neovascularization (CNV) plays an important role in pathogenesis of age-related macular degeneration (AMD), ocular histoplasmosis syndrome (OHS) and so on. However, mechanisms of CNV formation are n...Purpose: Choroidal neovascularization (CNV) plays an important role in pathogenesis of age-related macular degeneration (AMD), ocular histoplasmosis syndrome (OHS) and so on. However, mechanisms of CNV formation are not fully understood. The aim of this study is to investigate the correlation between expressions of CD105 and vascular endothelial growth factor (VEGF) in experimental laser-induced CNV in rats. Methods: CNV model was established by 532 nm laser photocoagulation in Brown-Norway rats. The expression of CD105 and VEGF in CNV was observed by immunohistochemistry at 3, 7, 14, 21, 28 and 56 days after laser photocoagulation. The image analysis was performed with the professional software of Image-Pro Plus. Results: Fluorescein angiography showed fluorescein leakage in CNV from days 7 to 56 after photocoagulation. VEGF expression was mainly observed in vascular endothelial cells, ganglion cells, inner nuclear layers and retinal pigment epithelial cells in normal retina and vascular endothelial cells in normal choroid of the rats. On day 3 after photocoagulation, VEGF began to express in laser-induced lesions. VEGF was strongly expressed in CNV after 7 days (P<0.05) and decreased after 14 days (P>0.05). CD105 was initially presented in CNV at 7 days and obviously expressed at 14 days after photocoagulation (P<0.05). Four weeks later, when angiogenesis tended toward inactive status, expression of CD105 was markedly decreased (P>0.05). There was notablely direct correlation between CD105-positive-microvessel density and positively semiquantitative scoring of VEGF in the CNV(r=0.989, P<0.01). Conclusions: There is direct correlation between the expression of CD105 and VEGF in the laser-induced CNV in rat. It suggests that CD105 and VEGF might participate in the new blood vessel formation and promote the growth of CNV.展开更多
AIM: To investigate the influence of hyperglycemia on the severity of choroidal neovascularization(CNV),especially the involvement of bone marrow-derived cells(BMCs) and underlying mechanisms.·METHODS: BMCs from ...AIM: To investigate the influence of hyperglycemia on the severity of choroidal neovascularization(CNV),especially the involvement of bone marrow-derived cells(BMCs) and underlying mechanisms.·METHODS: BMCs from firefly luciferase(Fluc)/green fluorescent protein(GFP) double transgenic mice were transplanted into C57BL/6J wide-type mice. The recipient mice were injected intraperitoneally with streptozotocin(STZ) daily for 5 consecutive days to induce diabetes mellitus(DM), followed by CNV laser photocoagulation.The BMCs recruitment in CNV exposed to hyperglycemia was firstly examined in Fluc/GFP chimeric mice by in vivo optical bioluminescence imaging(BLI) and in vitro Fluc assays. The CNV severity was evaluated by H&E staining and choroidal flatmount. The expression of vascular endothelial growth factor(VEGF) and stromal cell derived factor-1(SDF-1) was detected by Western blot.·RESULTS: BLI showed that the BMCs exerted dynamic effects in CNV model in Fluc/GFP chimeric mice exposed to hyperglycemia. The signal intensity of transplanted Fluc+GFP+BMCs in the DM chimeric mice was significantly higher than that in the control chimeric mice with CNV induction at days 5, 7, 14 and 21(121861.67 ±9948.81 vs 144998.33 ±13787.13 photons/second/cm2/sr for control and DM mice, P5d<0.05; 178791.67±30350.8 vs240166.67 ±22605.3, P7d<0.05; 124176.67 ±16253.52 vs196376.67 ±18556.79, P14d<0.05; 97951.60 ±10343.09 vs119510.00 ±14383.76, P21d<0.05), which was consistent with in vitro Fluc assay at day 7 [relative light units of Fluc(RLU1)], 215.00±52.05 vs 707.33±88.65, P <0.05; RLU1/relative light units of renilla luciferase(RLU2), 0.90 ±0.17 vs 1.83 ±0.17, P <0.05]. The CNVs in the DM mice were wider than those in the control group at days 5, 7, 14 and21(147.83±17.36 vs 220.33±20.17 μm, P5d<0.05; 212.17 ±24.63 vs 326.83 ±19.49, P7d<0.05; 163.17 ±18.24 vs265.17 ±20.55, P14d<0.05; 132.00 ±10.88 vs 205.33 ±12.98,P21d<0.05). The average area of CNV in the DM group was larger at 7d(20688.67±3644.96 vs 32218.00±4132.69 μm2,P <0.05). The expression of VEGF and SDF-1 was enhanced in the DM mice.·CONCLUSION: Hyperglycemia promots the vasculo-genesis of CNV, especially the contribution of BMCs,which might be triggered by VEGF and SDF-1 production.展开更多
AIM:To investigate the roles of integrins in choroidal neovascularization(CNV) and their associations with the stromal cell-derived factor-1(SDF-1)/CXCR4 axis.METHODS:CNV lesions were induced in mice using laser photo...AIM:To investigate the roles of integrins in choroidal neovascularization(CNV) and their associations with the stromal cell-derived factor-1(SDF-1)/CXCR4 axis.METHODS:CNV lesions were induced in mice using laser photocoagulation.After CNV induction,all animals were randomly assigned to:control,SDF-1,SDF-1+age-related macular degeneration(AMD) 3100(CXCR4 inhibitor),and SDF-1+ATN161(integrin α5β1 inhibitor) groups;their effects on CNV progression were observed using hematoxylin eosin(HE) staining,fundus fluorescein angiography(FFA) grading and optical coherence tomography(OCT),and their effects on CXCR4/integrin α5 expression were evaluated using Western blot and double immunofluorescence staining.Hypoxia-exposed endothelial cells(ECs) were used to simulate CNV in vitro,they were treated with SDF-1,combined with CXCR4 siRNA/AMD3100 or ATN161,and expression of integrin α5,cell migration and tube formation were analyzed.RESULTS:Integrin subunit α5 increased at 3 rd and 7 th day and decreased at 14 th day in CNV mice,with no significant change of β1-integrin.CXCR4 expression in CNV mice had persistent increase within 14 d after induction.SDF-1 treatment significantly promoted the CNV progression during 3-14 d.The mean CNV length in AMD3100 andATN161 group at day 7 was 270.13 and 264.23 μm in HE images,significantly lower than the mean length in SDF-1(345.70 μm) group.AMD3100 and ATN161 also significantly reduced thickness and leakage of CNV induced by SDF-1.Mean integrin α5 positive area in SDF-1 group reached 2.31×10~4 μm^2,significantly higher than control(1.25×104 μm^2),which decreased to 1.78×10~4 μm^2 after AMD3100 treatment.About 61.36% of ECs in CNV lesions expressed α5 in SDF-1 group,which significantly decreased to 43.12% after AMD3100 treatment.In vitro,integrin α5 peaked by 6 folds after 6 h of hypoxia exposure and CXCR4 gradually increased by up to 2.3 folds after 24 h of hypoxia.Approximately 25.12% of ECs expressed integrin α5 after SDF-1 stimulation,which decreased to 7.2%-9.5% after si-CXCR4 or AMD3100 treatment.ATN161 exerted an inhibitory effect comparable to that of si-CXCR4 on EC migration and tube formation in the presence of SDF-1.CONCLUSION:SDF-1/CXCR4 signaling induces integrin α5β1 expression in ECs to promote CNV.展开更多
AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein e...AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein endothelial cells(HUVECs)to induce hypoxia in vitro.Eight-week-old male C57 BL/6 J mice weighing 19-25 g were used for a choroidal neovascularization(CNV)model induced by laser photocoagulation in vivo.Expression levels of YAP,phosphorylated YAP,mesenchymal markers[αsmooth muscle actin(α-SMA),vimentin,and Snail],and endothelial cell markers(CD31 and zonula occludens 1)were measured by Western blotting,quantitative real-time PCR,and immunofluorescence microscopy.Small molecules YC-1(Lificiguat,a specific inhibitor of hypoxia-inducible factor 1α),CA3(CIL56,an inhibitor of YAP),and XMU-MP-1(an inhibitor of Hippo kinase MST1/2,which activates YAP)were used to explore the underlying mechanism.RESULTS:Co Cl2 increased expression of mesenchymal markers,decreased expression of endothelial cell markers,and enhanced the ability of primary HUVECs to proliferate and migrate.YC-1 suppressed hypoxia-induced endothelialto-mesenchymal transition(End MT).Moreover,hypoxia promoted total expression,inhibited phosphorylation,and enhanced the transcriptional activity of YAP.XMU-MP-1 enhanced hypoxia-induced End MT,whereas CA3 elicited the opposite effect.Expression of YAP,α-SMA,and vimentin were upregulated in the laser-induced CNV model.However,silencing of YAP by vitreous injection of small interfering RNA targeting YAP could reverse these changes.CONCLUSION:The findings reveal a critical role of the hypoxia-inducible factor-1α(HIF-1α)/YAP signaling axis in End MT and provide a new therapeutic target for treatment of subretinal fibrosis in AMD.展开更多
AIM: To investigate whether intravitreal injection of oxidized low-density lipoprotein(OxLDL) can promote laserinduced choroidal neovascularization(CNV) formation in mice and the mechanism involved, thereby to develop...AIM: To investigate whether intravitreal injection of oxidized low-density lipoprotein(OxLDL) can promote laserinduced choroidal neovascularization(CNV) formation in mice and the mechanism involved, thereby to develop a better animal model.METHODS: C57BL6/J mice were randomized into three groups. Immediately after CNV induction with 532 nm laser photocoagulation, 1.0 μL of OxLDL [100 μg/m L in phosphate-buffered saline(PBS)] was intravitreally injected, whereas PBS and the same volume low-density lipoprotein(LDL;100 μg/m L in PBS) were injected into the vitreous as controls. Angiogenic and inflammatory cytokines were measured by quantitative real-time polymerase chain reaction(q RT-PCR) and Western blotting(WB) after 5 d, and CNV severity was analyzed by choroid flat mount and immunofluorescence staining after 1wk. In vitro, retinal pigment epithelial(RPE) cell line(ARPE19) were treated with OxLDL(LDL as control) for 8 h. Angiogenic and inflammatory cytokine levels were measured. A specific inhibitor of lectin-like oxidized low-density lipoprotein receptor 1(LOX1) was used to evaluate the role of LOX1 in this process.RESULTS: At 7 d after intravitreal injection of 1 μL(100 μg/mL) OxLDL, T15-labeled OxLDL was mainly deposited around the CNV area, and the F4/80-labeled macrophages, the CD31-labeled vascular endothelial cells number and CNV area were increased. Meanwhile, WB and qR T-PCR results showed that vascular endothelial growth factor(VEGF), CC chemokine receptor 2(CCR2), interleukin-6(IL-6), IL-1β, and matrix metalloproteinase 9(MMP9) expressions were increased, which was supported by in vitro experiments in RPE cells. LOX1 inhibitors significantly reduced expressions of inflammatory factors IL-1β and VEGF. CONCLUSION: A modified laser-induced CNV animal model is established with intravitreal injection of 1 μL(100 μg/mL) of OxLDL at 7 d, which at least partially through LOX1. This animal model can be used as a simple model for studying the role of OxLDL in age-related macular degeneration.展开更多
AIM: To examine the effect of intravitreal adenoviral vector-mediated tristetraprolin(Ad-TTP) on VEGF m RNA expression in a rat model of laser-induced choroidal neovascularization.METHODS: Ad-TTP was prepared using a ...AIM: To examine the effect of intravitreal adenoviral vector-mediated tristetraprolin(Ad-TTP) on VEGF m RNA expression in a rat model of laser-induced choroidal neovascularization.METHODS: Ad-TTP was prepared using a commercial kit. Retinal laser-induced photocoagulation(10 spots per eye) was performed on rats in this experimental choroidal neovascularization(CNV) model. Rats were divided into four groups: control(single intravitreal injection of balanced salt solution, n =10), laser-induced CNV(photocoagulation only, n =20), laser-induced CNV plus Ad-TTP injection(photocoagulation plus a single intravitreal Ad-TTP injection, n =20) and Ad-TTP injection only(n =10). Changes in choroidal morphology were evaluated in ten rats in the laser only and the laser plus Ad-TTP groups. Two weeks after laser injury, the size of CNV was calculated by perfusion with high-molecular-weight fluorescein isothiocyanate(FITC)-dextran. VEGF m RNA expression in retina-choroid tissue from ten rats in each group was measured by reverse transcription polymerase chain reaction(RT-PCR). RESULTS: Two weeks after treatment, the area of laser-induced CNV was reduced by approximately 60% in the rats given the Ad-TTP injection compared with that in the laser-only group. There was a tendency toward decreased VEGF m RNA expression in the Ad-TTP injection groups.CONCLUSION: A single intravitreal injection of Ad-TTP significantly suppressed CNV size in this experimental laser-induced CNV model. Ad-TTP injection also decreased VEGF m RNA expression compared with that inthe laser-induced CNV group. The present study is meaningful as the first study to investigate the effect of tristetraprolin delivered via intravitreal injection.展开更多
AIM:To study the effects of cytokeratin 17(CK17) on sodium iodate(NalO_3) induced rat retinal pigment epithelium(RPE) degeneration,laser induced rat choroidal neovascularization(CNV),and oxidative stress of human reti...AIM:To study the effects of cytokeratin 17(CK17) on sodium iodate(NalO_3) induced rat retinal pigment epithelium(RPE) degeneration,laser induced rat choroidal neovascularization(CNV),and oxidative stress of human retinal pigment epithelium cells(ARPE-19) and human umbilical vein endothelial cell(HUVEC).· METHODS:Thirty 8-week-old male Brown Norway rats were randomly divided into 3 groups,10 rats in control group treated with solvent alone;10 rats in NalO_3 group treated with solvent and 35 mg/kg NalO_3 injection through hypoglossal vein and 10 rats in CK17+NalO_3group treated with 1%CK17 eye drop 3 times a day for1 wk before and 4wk after NalO_3 injection.RPE function was measured with c-wave of electroretinogram(ERG).Another 20 rats were randomly divided into 2 groups.Of them 10 rats in CK17 group were anesthetized to receive Nd:YAG laser and given 1%CK17 eye drop before same as above;10 rats in control were received Nd:YAG and treated with solvent.The development of choroidal neovascularization(CNV) was determined by fundus fluorescein angiography(FFA) performed on 4wk after laser.Methylthiazoly tetrazolium(MTT) assay was used to study effect of CK17 on various oxidants induced injury in ARPE-19 and HUVEC in vitro.· RESULTS:Four weeks after NalO_3 injection,the cwave amplitude of ERG was 0.393±0.02 V in the control group,0.184 ±0.018 V in NalO_3 group and 0.3±0.01 V in CK17+NalO_3 group.There was a significant reversal of the c-wave by CK17 as compared to NalO_3 group(P<0.01).Four weeks after laser,the size of the CNV lesion was2.57±0.27 mm^2 in control group and 1.64±0.08 mm_2 in CK17 group.The lesion size significantly diminished in CK17 group(P<0.01).The in vitro results showed CK17 also reversed the various oxidants induced injuries in ARPE-19 at the dose of 100 μg/mL and enhanced the injury in HUVECs at different concentrations.CONCLUSION:CK17 can significantly protect RPE from NalO_3 induced degeneration in vivo and in vitro and also could reverse the various oxidants induced injuries in vitro.It inhibits the development of CNV in rat model,interfered with vascular endothelial cell proliferation in vitro.展开更多
AIM:To determine the effects of safranal on choroidal neovascularization(CNV)and oxidative stress damage of human choroidal microvascular endothelial cells(HCVECs)and its possible mechanisms.METHODS:Forty-five rats we...AIM:To determine the effects of safranal on choroidal neovascularization(CNV)and oxidative stress damage of human choroidal microvascular endothelial cells(HCVECs)and its possible mechanisms.METHODS:Forty-five rats were used as a laser-induced CNV model for testing the efficacy and safety of safranal(0.5 mg/kg·d,intraperitoneally)on CNV.CNV leakage on fluorescein angiography(FA)and CNV thickness on histology was compared.HCVECs were used for a H_(2)O_(2)-induced oxidative stress model to test the effect of safranal in vitro.MTT essay was carried to test the inhibition rate of safranal on cell viability at different concentrations.Tube formation was used to test protective effect of safranal on angiogenesis at different concentrations.mRNA transcriptome sequencing was performed to find the possible signal pathway.The expressions of different molecules and their phosphorylation level were validated by Western blotting.RESULTS:On FA,the average CNV leakage area was 0.73±0.49 and 0.31±0.11 mm^(2)(P=0.012)in the control and safranal-treated group respectively.The average CNV thickness was 127.4±18.75 and 100.6±17.34μm(P=0.001)in control and safranal-treated group.Under the condition of oxidative stress,cell proliferation was inhibited by safranal and inhibition rates were 7.4%-35.4%at the different concentrations.For tube formation study,the number of new branches was 364 in control group and 35,42,and 17 in 20,40,and 80μg/mL safranal groups respectively(P<0.01).From the KEGG pathway bubble graph,the PI3K-AKT signaling pathway showed a high gene ratio.The protein expression was elevated of insulin receptor substrate(IRS)and the phosphorylation level of PI3K,phosphoinositide-dependent protein kinase 1/2(PDK1/2),AKT and Bcl-2 associated death promoter(BAD)was also elevated under oxidative stress condition but inhibited by safranal.CONCLUSION:Safranal can inhibit CNV both in vivo and in vitro,and the IRS-PI3K-PDK1/2-AKT-BAD signaling pathway is involved in the pathogenesis of CNV.展开更多
AIM:To observe the long-term clinical efficacy of intravitreal injections of conbercept,a novel vascular growth factor inhibitor,for the treatment of pathological myopia choroidal neovascularization(PM-CNV).METHODS:A ...AIM:To observe the long-term clinical efficacy of intravitreal injections of conbercept,a novel vascular growth factor inhibitor,for the treatment of pathological myopia choroidal neovascularization(PM-CNV).METHODS:A total of 67 eyes(from 67 patients;mean age,54.90±12.7y)with PM-CNV were retrospectively researched.Based on the different schemes used for the administration of the drug,the patients were divided into two groups:group A(n=35;average age,53.31±13.6y;average diopter,9.25±1.72 D),which received only one injection of pro re nata(PRN;1+PRN regimen),and group B(n=32;average age,56.49±11.8y;average diopter,9.63±2.24 D),which received one injection per month for 3mo(3+PRN regimen).Best-corrected visual acuity(BCVA)analysis,intraocular pressure(IOP)examination,slit-lamp microscopy,fundus examination and optical coherence tomography were per formed at each follow-up.The recurrence and treatment times of CNV were recorded.The patients were followed up for at least 12mo.RESULTS:The BCVA was increased in 29 eyes(82.9%)in group A and 30 eyes(93.75%)in group B;no increase or decrease was observed in 6(17.1%)and 2(6.25%)eyes in groups A and B,respectively.The BCVA(log MAR)values before treatment(0.67±0.48 and 0.71±0.56)were significantly higher than those 12mo after treatment(0.31±0.26 and 0.33±0.17)in groups A and B,respectively(P<0.05).The mean central macular thickness(CMT)values had significantly decreased from 346.49±65.99 and 360.10±82.31μm at baseline to 257.29±40.47 and 251.97±48.26μm in groups A and B,respectively,after 12mo of treatment.A total of 21 eyes in group A needed reinjection(60%;average number of injections,2.51±0.98);the corresponding values in group B were 6 eyes(18.75%;average number of injections,3.74±1.22).There were no adverse ocular and systemic complications during the treatment and follow-up.CONCLUSION:Intravitreal injection of conbercept with 1+PRN or 3+PRN improve the visual acuity,reduce macular edema and reduce the level of CMT in patients with PM-CNV.The 3+PRN regimen demonstrates a lower recurrence rate of CNV than the 1+PRN regimen,but requires more treatment.However,both treatment regimens demonstrate long-term safety and efficacy for the treatment of PM-CNV.展开更多
AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was...AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was treated with celastrol to evaluate this natural compound's impact on CNV.Western blot was applied to explore the possible mechanism for it.For in vitro assay,triplicate for each group was repeated at least three times.For in vivo assay,each group contains 5 mice.RESULTS:Celastrol supressed tube formation and aortic ring sprout neovascularization.In vitro assay exhibited that celastrol inhibiting vascular endothelial growth factor(VEGF)-induced proliferation and migration of human umbilical vein endothelial cells and human choroidal endothelial cells,and by blocking VEGF signaling.Furthermore,intraperitoneal administration of celastrol significantly reduced the area of laser-induced CNV in an in vivo mouse model.By day 14,the area of CNV had decreased by 49.15%and 80.26%in the 0.1 mg/kg celastrol-treated group(n=5)and in the 0.5 mg/kg celastrol treated group(n=5),respectively,compared to the vehicle-treated group(n=5).CONCLUSION:Celastrol inhibits CNV by inhibiting VEGF-induced proliferation and migration of vascular endothelial cells,indicating that celastrol is a potent,natural therapeutic compound for the prevention of CNV.展开更多
AIM:To compare choroidal neovascularization(CNV)lesion measurements obtained by in vivo imaging modalities,with whole mount histological preparations stained with isolectin GS-IB4,using a murine laser-induced CNV mode...AIM:To compare choroidal neovascularization(CNV)lesion measurements obtained by in vivo imaging modalities,with whole mount histological preparations stained with isolectin GS-IB4,using a murine laser-induced CNV model.METHODS:B6 N.Cg-Tg(Csf1 r-EGFP)1 Hume/J heterozygous adult mice were subjected to laser-induced CNV and were monitored by fluorescein angiography(FA),multicolor(MC)fundus imaging and optical coherence tomography angiography(OCTA)at day 14 after CNV induction.Choroidalretinal pigment epithelium(RPE)whole mounts were prepared at the end of the experiment and were stained with isolectin GS-IB4.CNV areas were measured in all different imaging modalities at day 14 after CNV from three independent raters and were compared to choroidal-RPE whole mounts.Intraclass correlation coefficient(ICC)type 2(2-way random model)and its 95%confidence intervals(CI)were calculated to measure the correlation between different raters’measurements.Spearman’s rank correlation coefficient(Spearman’s r)was calculated for the comparison between FA,MC and OCTA data and histology data.RESULTS:FA(early and late)and MC correlates well with the CNV measurements ex vivo with FA having slightly better correlation than MC(FA early Spearman’s r=0.7642,FA late Spearman’s r=0.7097,and MC Spearman’s r=0.7418),while the interobser ver reliability was good for both techniques(FA early ICC=0.976,FA late ICC=0.964,and MC ICC=0.846).In contrast,OCTA showed a poor correlation with ex vivo measurements(Spearman’s r=0.05716)and high variability between different raters(ICC=0.603).CONCLUSION:This study suggests that FA and MC imaging could be used for the evaluation of CNV areas in vivo while caution must be taken and comparison studies should be performed when OCTA is employed as a CNV monitoring tool in small rodents.展开更多
AIM:To analyse macular microvascular alterations in myopic choroidal neovascularization(m CNV)and the efficiency of anti-vascular endothelial growth factor(antiVEGF)therapy for m CNV by optical coherence tomography an...AIM:To analyse macular microvascular alterations in myopic choroidal neovascularization(m CNV)and the efficiency of anti-vascular endothelial growth factor(antiVEGF)therapy for m CNV by optical coherence tomography angiography(OCTA).METHODS:A total of 123 patients were included in this retrospective study,divided into m CNV group,high myopia(HM)group,and normal group at the Affiliated Eye Hospital of Wenzhou Medical University from January 2017 to January 2019.Superficial vessel density,deep capillary density,foveal avascular zone(FAZ)area,A-circularity index(AI)and vessel density around the 300μm width of the FAZ region density(FD)and the area of choroidal neovascularization(CNV)lesion(only for m CNV group)were measured on 3×3 mm2 OCTA images.FAZ area was corrected for axial length.Central macular thickness(CMT)was measured on OCT in m CNV group.Compared the parameters on OCTA of 3 groups and pre-anti-VEGF and post-anti-VEGF at 1,2,3,and 6 mo follow-up in m CNV group.RESULTS:There were significant differences among 3 groups in superficial vessel density,deep capillary density and FD(P<0.05).FAZ area in HM group was smaller than normal group(P<0.05),but there was no significant difference between m CNV group and the other two group.AI increased in m CNV group(P<0.05).The mean CMT,area and flow area of CNV lesion decreased after treatment(P<0.05),while vessel density and FAZ didn’t change.The mean CMT,area and flow area of CNV lesion statistically decreased after anti-VEGF treatment in m CNV group(P<0.05),while superficial vessel density,deep capillary density and FAZ area,AI and FD didn’t change.The mean reduction ratio of lesions was 50.32%(7.07%to 100%).Lesion regression 100%was observed in 2 cases(4.88%).There was a negative correlation between the CNV lesion area and reduction ratio(r=-0.380,P=0.042)and the flow lesion area and reduction ratio(r=-0.402,P=0.030).CONCLUSION:Macular vessel density decreases,FAZ turns smaller and more irregular in m CNV eyes.AntiVEGF therapy is efficient for m CNV without affecting vessel density and FAZ,but it is unable to completely eliminate CNV lesions in most cases.The bigger m CNV lesions have lower reduction ratio.展开更多
Dear Editor,I am Cheolmin Yun,from the Department of Ophthalmology,Korea University College of Medicine.I write to present a case report of a female patient with a myopic patient suffering from atrophic choroidal neov...Dear Editor,I am Cheolmin Yun,from the Department of Ophthalmology,Korea University College of Medicine.I write to present a case report of a female patient with a myopic patient suffering from atrophic choroidal neovascularization(CNV)and a full thickness macular hole(FTMH),who was treated with an intravitreal展开更多
AIM:To investigate whether anti-placental growth factor(PGF) can inhibit subretinal fibrosis and whether this effect is mediated by the inhibitory effect of PGF on epithelial-mesenchymal transition(EMT) of retinal pig...AIM:To investigate whether anti-placental growth factor(PGF) can inhibit subretinal fibrosis and whether this effect is mediated by the inhibitory effect of PGF on epithelial-mesenchymal transition(EMT) of retinal pigment epithelial(RPE) cells.METHODS:Subretinal fibrosis model was established in laser induced choroidal neovascularization(CNV) mice on day 21 after laser photocoagulation.Immunofluorescence staining(IFS) of cryosections and enzyme-linked immunosorbent assay(ELISA) were used to detect the expression of PGF.IFS of whole choroidal flat-mounts was used to detect the degree of subretinal fibrosis.IFS of cryosections and ELISA were used to detect the expression of EMT related indicators in subretinal fibrosis lesions.RESULTS:The expression of PGF protein in subretinal fibrosis lesions was significantly up-regulated(P<0.05),and mainly co-stained with pan-cytokeratin labeled RPE cells.Intravitreal injection of anti-PGF neutralizing antibody reduced the area of subretinal fibrosis and the ratio of fibrotic/angiogenic area significantly at the concentrations of 0.25,0.5,1.0,and 2.0 μg/μL(all P<0.05).The expression of E-cadherin in the local RPE cells decreased,while α-SMA increased significantly in subretinal fibrosis lesions,and the application of anti-PGF neutralizing antibody could reverse these changes(P<0.05).CONCLUSION:The expression of PGF is up-regulated in the lesion site of subretinal fibrosis and mainly expressed in RPE cells.Intravitreal injection of anti-PGF neutralizing antibody can significantly inhibit the degree of subretinal fibrosis in CNV mice,and this effect may be mediated by the inhibition of PGF on EMT of RPE cells.展开更多
AIM: To investigate the clinical characteristics and genetic features of a Bietti crystalline dystrophy(BCD) proband in a Chinese family.METHODS: A Chinese female diagnosed with BCD complicated by bilateral choroidal ...AIM: To investigate the clinical characteristics and genetic features of a Bietti crystalline dystrophy(BCD) proband in a Chinese family.METHODS: A Chinese female diagnosed with BCD complicated by bilateral choroidal neovascularization(CNV) and her parents underwent complete ophthalmic examinations, including fundus autofluorescence(AF), fundus photography(FP), fundus fluorescein angiography(FFA), visual field testing, full-field electroretinography(ERG), optical coherence tomography(OCT) and optical coherence tomography angiography(OCTA). The sequencing of the CYP4 V2 gene was performed to the whole family.RESULTS: Bilateral tiny glittering crystal-like deposits and differing extent of atrophy of the retinal pigment epithelium(RPE) were found in the posterior pole of her fundus. The diffuse hypo-fluorescence shown on AF images and window defects shown on FFA both indicated the atrophy of the RPE and choriocapillaris. OCT showed the thinning of the RPE and choriocapillaris layer, ellipsoid zone(EZ) band defect and CNV in both eyes. OCTA images proofed bilateral type 2 CNV. The visual field test showed central and paracentral scotoma. ERG showed a slightly decreased b-wave in scotopic ERG. Gene sequencing identified three mutations of the CYP4 V2 gene, c.802_807 del, c.810 del T, and c.1388 G>A. The mutation c.1388 G>A was a novel substitution mutation.CONCLUSION: The novel mutation c.1388 G>A may be a possible cause that could induce the clinical phenotype of BCD.展开更多
AIM:To investigate therapeutic effects of traditional Chinese medicine formulations,Hexuemingmu(HXMM)on laser-induced choroidal neovascularization(CNV)and followup effect in mice.METHODS:C57 BL/6 mice of 8-week-old we...AIM:To investigate therapeutic effects of traditional Chinese medicine formulations,Hexuemingmu(HXMM)on laser-induced choroidal neovascularization(CNV)and followup effect in mice.METHODS:C57 BL/6 mice of 8-week-old were used and CNV was induced with 577 nm laser photocoagulation.Animals were randomly divided into groups and different doses of HXMM were administered daily.One,four,and eight weeks after the intervention,the electroretinogram(ERG),fundus fluorescence angiography,choroidal flat mount and immunofluorescence staining were preformed to evaluate the function and CNV formation.The expression levels of angiogenic proteins were determined by Western blotting and immunofluorescence staining.An analysis of variance and Kruskal-Wallis test were used to test the differences among the groups.RESULTS:The results showed that HXMM effectively increased amplitude of ERG of mice(P<0.05),alleviated fundus CNV leakage(P<0.05),and reduced the area of neovascularization and the expression of angiogenic proteins(P<0.05)after laser-induced CNV.CONCLUSION:HXMM can protect the retinal function of mice after laser-induced CNV,and inhibit the CNV development.展开更多
Dear Editor,My name is Georgios Tsokolas and I am currently working as Medical Retina Research Fellow at the Eye Unit of Southampton General Hospital in United Kingdom.I am writing this letter to present three case se...Dear Editor,My name is Georgios Tsokolas and I am currently working as Medical Retina Research Fellow at the Eye Unit of Southampton General Hospital in United Kingdom.I am writing this letter to present three case series of choroidal neovascular(CNV)membrane after macular surgery。展开更多
文摘AIM:To evaluate effects of endogenous dopamine induced by low concentration atropine eye drops on choroidal neovascularization(CNV)in high myopia mice.METHODS:The C57BL/6J mice were deprived of the right eye for 4wk,and the high myopia was diagnosed by optometry,the diopter was less than-6.00 D,and CNV was induced by 532 nm laser.The changes of dopamine D1 receptor(DRD1),dopamine D2 receptor(DRD2),and vascular endothelial growth factor A(VEGFA)were detected by Western blot technology at 0.5,1,2h,and 7d after 0.01%,0.05%,and 0.1%atropine eye drops,respectively,the area of CNV was measured.RESULTS:Significant increases were observed on the expression of DRD2 in mouse high myopia model at 0.5,1,2h,7d with 0.05%and 0.1%atropine eye drops(P<0.05).Significant decreases were observed on the expression of DRD1 and VEGFA in mouse high myopia model at 0.5,1,2h,7d with 0.05%and 0.1%atropine eye drops(P<0.05).The area of CNV induced by laser in the drug-treated group was significantly smaller than that in the control group,and the higher the concentration,the more significant the inhibitory effect(P<0.05).CONCLUSION:The 0.01%,0.05%,0.1%atropine eye drops can decrease the level of VEGFA and inhibit high myopia CNV indirectly by up-regulating the level of DRD2 and down-regulating the level of DRD1,and the effect of 0.05%and 0.1%atropine eye drops is more significant.
基金Supported by 2023 Research Fund of Aier Ophthalmology Research Institute(No.AEI202310LC01).
文摘AIM:To evaluate the efficacy of intravitreal injection of anti-vascular endothelial growth factor(anti-VEGF),photodynamic therapy(PDT),and laser treatment(LT)for anatomical and functional improvement in myopic choroidal neovascularization(mCNV)patients.METHODS:Two researchers independently searched PubMed,Cochrane Library,Web of Science,and other databases to screen studies comparing best-corrected vision acuity(BCVA)and foveal center thickness(FCT)changes after mCNV treatment.Post-treatment chorioretinal atrophy(CRA)is a secondary outcome indicator.The retrieval time limit is from the database construction to January 30,2023.RESULTS:A total of 1072 eyes in 16 articles were included.In the RCTs,intravitreal bevacizumab(IVB)and intravitreal ranibizumab(IVR)were superior to PDT(MD=0.18,95%CI:0.02,0.40,MD=0.18,95%CI:0.01,0.42)in improving BCVA of mCNV patients(P<0.05).The relative effectiveness in improving BCVA,from high to low,appeared to be IVR,intravitreal aflibercept(IVA),IVB,LT,PDT,and sham first followed by IVA(Sham/IVA).While improving the FCT from high to low was IVA,IVR,IVB,PDT.In retrospective studies,the results of BCVA after long-term treatment showed that all the therapeutic effects from high to low was IVA,intravitreal conbercept(IVC),IVR,IVB,IVB/IVR,PDT with IVB/IVR,PDT.The effect of improving FCT was IVA,IVR,IVC,PDT,and IVB from high to low.And in the effects of improving CRA,the IVB appeared to be higher than IVR,while the PDT was the smallest,but none of the differences in the results were statistically significant.CONCLUSION:Anti-VEGF has the best effect on longterm vision improvement in mCNV patients,using IVB or IVR alone to treat mCNV may be better than IVB or IVR combined with PDT.There is no significant difference in the improvement of visual acuity,macular edema,and CRA in mCNV patients treated with any different anti-VEGF drugs.
基金Supported by the National Natural Science Foundation of China(No.81703134,No.81770952)Henan Province Nature Science Foundation of China(No.162300410296)Hunan Province Nature Science Foundation of China(No.2018JJ3772)。
文摘AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue tetrazolium bromide(M一)assay.Effects of RSV on choroidal endothelial cell(CEC)migration were evaluated with a modified Boyden chamber assay,while tube formation was evaluated in a 2-D gel assay.CNV was induced by laser photocoagulation in mice.The effects of intravitreal injection of RSV on CNV development were evaluated by fluorescein angiography(FA),confocal analysis of isolectin B4 labeled choroidal flat mounts,and histologic examination of CNV membranes.Immunostaining was used to analyze the expression and phosphorylation of vascular endothelial growth factor receptor 2(VEGFR2).RESULTS:No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 pmol/L(P>0.05).RSV inhibited vascular endothelial growth factor(VEGF)-induced CEC migration(P<0.05)and tube formation(P<0.05)invitro.Furthermore,intravitrealinjectionof RSV significantly inhibited laser induced CNV formation in mice.The FA leakage,CNV volume and CNV area analysis revealed that there were 41%,45%,and 58%reduction in RSV-treated eyes(1.691±0.1032,178163±78623μm^3 and 6508±619.0μm^2,respectively)compared with those in control(2.724±0.08447,379676±98382μm3and16576±2646μm^2,respectively;P<0.05).Phospho-VEGFR2expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control(P<0.05).CONCLUSION:Intravitreal injection of RSV exerts an inhibitory effect on CNV,which may through suppressing endothelial cell migration,tube formation and VEGFR2 phosphorylation.
基金Supported by National Natural Science Foundation ( No.30371516) , the Scientific Research Foundation for the ReturnedOverseas Chinese Scholars, State Education Ministry, China ( 2004) , Natural Science Foundation of Shanxi Province ( No.2004C246) and Scientific and Technological Innovation Foundation of Xijing Hospital ( No.XJCX04M003)
文摘Purpose: Choroidal neovascularization (CNV) plays an important role in pathogenesis of age-related macular degeneration (AMD), ocular histoplasmosis syndrome (OHS) and so on. However, mechanisms of CNV formation are not fully understood. The aim of this study is to investigate the correlation between expressions of CD105 and vascular endothelial growth factor (VEGF) in experimental laser-induced CNV in rats. Methods: CNV model was established by 532 nm laser photocoagulation in Brown-Norway rats. The expression of CD105 and VEGF in CNV was observed by immunohistochemistry at 3, 7, 14, 21, 28 and 56 days after laser photocoagulation. The image analysis was performed with the professional software of Image-Pro Plus. Results: Fluorescein angiography showed fluorescein leakage in CNV from days 7 to 56 after photocoagulation. VEGF expression was mainly observed in vascular endothelial cells, ganglion cells, inner nuclear layers and retinal pigment epithelial cells in normal retina and vascular endothelial cells in normal choroid of the rats. On day 3 after photocoagulation, VEGF began to express in laser-induced lesions. VEGF was strongly expressed in CNV after 7 days (P<0.05) and decreased after 14 days (P>0.05). CD105 was initially presented in CNV at 7 days and obviously expressed at 14 days after photocoagulation (P<0.05). Four weeks later, when angiogenesis tended toward inactive status, expression of CD105 was markedly decreased (P>0.05). There was notablely direct correlation between CD105-positive-microvessel density and positively semiquantitative scoring of VEGF in the CNV(r=0.989, P<0.01). Conclusions: There is direct correlation between the expression of CD105 and VEGF in the laser-induced CNV in rat. It suggests that CD105 and VEGF might participate in the new blood vessel formation and promote the growth of CNV.
基金Supported by the National Natural Science Foundation of China(No.81070748,No.81200708)National Basic Research Program of China(973 Program)
文摘AIM: To investigate the influence of hyperglycemia on the severity of choroidal neovascularization(CNV),especially the involvement of bone marrow-derived cells(BMCs) and underlying mechanisms.·METHODS: BMCs from firefly luciferase(Fluc)/green fluorescent protein(GFP) double transgenic mice were transplanted into C57BL/6J wide-type mice. The recipient mice were injected intraperitoneally with streptozotocin(STZ) daily for 5 consecutive days to induce diabetes mellitus(DM), followed by CNV laser photocoagulation.The BMCs recruitment in CNV exposed to hyperglycemia was firstly examined in Fluc/GFP chimeric mice by in vivo optical bioluminescence imaging(BLI) and in vitro Fluc assays. The CNV severity was evaluated by H&E staining and choroidal flatmount. The expression of vascular endothelial growth factor(VEGF) and stromal cell derived factor-1(SDF-1) was detected by Western blot.·RESULTS: BLI showed that the BMCs exerted dynamic effects in CNV model in Fluc/GFP chimeric mice exposed to hyperglycemia. The signal intensity of transplanted Fluc+GFP+BMCs in the DM chimeric mice was significantly higher than that in the control chimeric mice with CNV induction at days 5, 7, 14 and 21(121861.67 ±9948.81 vs 144998.33 ±13787.13 photons/second/cm2/sr for control and DM mice, P5d<0.05; 178791.67±30350.8 vs240166.67 ±22605.3, P7d<0.05; 124176.67 ±16253.52 vs196376.67 ±18556.79, P14d<0.05; 97951.60 ±10343.09 vs119510.00 ±14383.76, P21d<0.05), which was consistent with in vitro Fluc assay at day 7 [relative light units of Fluc(RLU1)], 215.00±52.05 vs 707.33±88.65, P <0.05; RLU1/relative light units of renilla luciferase(RLU2), 0.90 ±0.17 vs 1.83 ±0.17, P <0.05]. The CNVs in the DM mice were wider than those in the control group at days 5, 7, 14 and21(147.83±17.36 vs 220.33±20.17 μm, P5d<0.05; 212.17 ±24.63 vs 326.83 ±19.49, P7d<0.05; 163.17 ±18.24 vs265.17 ±20.55, P14d<0.05; 132.00 ±10.88 vs 205.33 ±12.98,P21d<0.05). The average area of CNV in the DM group was larger at 7d(20688.67±3644.96 vs 32218.00±4132.69 μm2,P <0.05). The expression of VEGF and SDF-1 was enhanced in the DM mice.·CONCLUSION: Hyperglycemia promots the vasculo-genesis of CNV, especially the contribution of BMCs,which might be triggered by VEGF and SDF-1 production.
基金Supported by the National Natural Science Foundation of China(No.81770936No.81570856+2 种基金No.81670863No.81500748No.81370020)
文摘AIM:To investigate the roles of integrins in choroidal neovascularization(CNV) and their associations with the stromal cell-derived factor-1(SDF-1)/CXCR4 axis.METHODS:CNV lesions were induced in mice using laser photocoagulation.After CNV induction,all animals were randomly assigned to:control,SDF-1,SDF-1+age-related macular degeneration(AMD) 3100(CXCR4 inhibitor),and SDF-1+ATN161(integrin α5β1 inhibitor) groups;their effects on CNV progression were observed using hematoxylin eosin(HE) staining,fundus fluorescein angiography(FFA) grading and optical coherence tomography(OCT),and their effects on CXCR4/integrin α5 expression were evaluated using Western blot and double immunofluorescence staining.Hypoxia-exposed endothelial cells(ECs) were used to simulate CNV in vitro,they were treated with SDF-1,combined with CXCR4 siRNA/AMD3100 or ATN161,and expression of integrin α5,cell migration and tube formation were analyzed.RESULTS:Integrin subunit α5 increased at 3 rd and 7 th day and decreased at 14 th day in CNV mice,with no significant change of β1-integrin.CXCR4 expression in CNV mice had persistent increase within 14 d after induction.SDF-1 treatment significantly promoted the CNV progression during 3-14 d.The mean CNV length in AMD3100 andATN161 group at day 7 was 270.13 and 264.23 μm in HE images,significantly lower than the mean length in SDF-1(345.70 μm) group.AMD3100 and ATN161 also significantly reduced thickness and leakage of CNV induced by SDF-1.Mean integrin α5 positive area in SDF-1 group reached 2.31×10~4 μm^2,significantly higher than control(1.25×104 μm^2),which decreased to 1.78×10~4 μm^2 after AMD3100 treatment.About 61.36% of ECs in CNV lesions expressed α5 in SDF-1 group,which significantly decreased to 43.12% after AMD3100 treatment.In vitro,integrin α5 peaked by 6 folds after 6 h of hypoxia exposure and CXCR4 gradually increased by up to 2.3 folds after 24 h of hypoxia.Approximately 25.12% of ECs expressed integrin α5 after SDF-1 stimulation,which decreased to 7.2%-9.5% after si-CXCR4 or AMD3100 treatment.ATN161 exerted an inhibitory effect comparable to that of si-CXCR4 on EC migration and tube formation in the presence of SDF-1.CONCLUSION:SDF-1/CXCR4 signaling induces integrin α5β1 expression in ECs to promote CNV.
基金National Natural Science Foundation of China(No.81970817,No.81873680)。
文摘AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein endothelial cells(HUVECs)to induce hypoxia in vitro.Eight-week-old male C57 BL/6 J mice weighing 19-25 g were used for a choroidal neovascularization(CNV)model induced by laser photocoagulation in vivo.Expression levels of YAP,phosphorylated YAP,mesenchymal markers[αsmooth muscle actin(α-SMA),vimentin,and Snail],and endothelial cell markers(CD31 and zonula occludens 1)were measured by Western blotting,quantitative real-time PCR,and immunofluorescence microscopy.Small molecules YC-1(Lificiguat,a specific inhibitor of hypoxia-inducible factor 1α),CA3(CIL56,an inhibitor of YAP),and XMU-MP-1(an inhibitor of Hippo kinase MST1/2,which activates YAP)were used to explore the underlying mechanism.RESULTS:Co Cl2 increased expression of mesenchymal markers,decreased expression of endothelial cell markers,and enhanced the ability of primary HUVECs to proliferate and migrate.YC-1 suppressed hypoxia-induced endothelialto-mesenchymal transition(End MT).Moreover,hypoxia promoted total expression,inhibited phosphorylation,and enhanced the transcriptional activity of YAP.XMU-MP-1 enhanced hypoxia-induced End MT,whereas CA3 elicited the opposite effect.Expression of YAP,α-SMA,and vimentin were upregulated in the laser-induced CNV model.However,silencing of YAP by vitreous injection of small interfering RNA targeting YAP could reverse these changes.CONCLUSION:The findings reveal a critical role of the hypoxia-inducible factor-1α(HIF-1α)/YAP signaling axis in End MT and provide a new therapeutic target for treatment of subretinal fibrosis in AMD.
基金Supported by the National Natural Science Foundation of China (No.81470654)Science and Technology Plan of Natural Science Foundation of Shaanxi Province (No.2019SF-047)。
文摘AIM: To investigate whether intravitreal injection of oxidized low-density lipoprotein(OxLDL) can promote laserinduced choroidal neovascularization(CNV) formation in mice and the mechanism involved, thereby to develop a better animal model.METHODS: C57BL6/J mice were randomized into three groups. Immediately after CNV induction with 532 nm laser photocoagulation, 1.0 μL of OxLDL [100 μg/m L in phosphate-buffered saline(PBS)] was intravitreally injected, whereas PBS and the same volume low-density lipoprotein(LDL;100 μg/m L in PBS) were injected into the vitreous as controls. Angiogenic and inflammatory cytokines were measured by quantitative real-time polymerase chain reaction(q RT-PCR) and Western blotting(WB) after 5 d, and CNV severity was analyzed by choroid flat mount and immunofluorescence staining after 1wk. In vitro, retinal pigment epithelial(RPE) cell line(ARPE19) were treated with OxLDL(LDL as control) for 8 h. Angiogenic and inflammatory cytokine levels were measured. A specific inhibitor of lectin-like oxidized low-density lipoprotein receptor 1(LOX1) was used to evaluate the role of LOX1 in this process.RESULTS: At 7 d after intravitreal injection of 1 μL(100 μg/mL) OxLDL, T15-labeled OxLDL was mainly deposited around the CNV area, and the F4/80-labeled macrophages, the CD31-labeled vascular endothelial cells number and CNV area were increased. Meanwhile, WB and qR T-PCR results showed that vascular endothelial growth factor(VEGF), CC chemokine receptor 2(CCR2), interleukin-6(IL-6), IL-1β, and matrix metalloproteinase 9(MMP9) expressions were increased, which was supported by in vitro experiments in RPE cells. LOX1 inhibitors significantly reduced expressions of inflammatory factors IL-1β and VEGF. CONCLUSION: A modified laser-induced CNV animal model is established with intravitreal injection of 1 μL(100 μg/mL) of OxLDL at 7 d, which at least partially through LOX1. This animal model can be used as a simple model for studying the role of OxLDL in age-related macular degeneration.
基金Supported by Biomedical Research Institute Fund (GNUHBRIF-2013-0002) from the Gyeongsang National University Hospital
文摘AIM: To examine the effect of intravitreal adenoviral vector-mediated tristetraprolin(Ad-TTP) on VEGF m RNA expression in a rat model of laser-induced choroidal neovascularization.METHODS: Ad-TTP was prepared using a commercial kit. Retinal laser-induced photocoagulation(10 spots per eye) was performed on rats in this experimental choroidal neovascularization(CNV) model. Rats were divided into four groups: control(single intravitreal injection of balanced salt solution, n =10), laser-induced CNV(photocoagulation only, n =20), laser-induced CNV plus Ad-TTP injection(photocoagulation plus a single intravitreal Ad-TTP injection, n =20) and Ad-TTP injection only(n =10). Changes in choroidal morphology were evaluated in ten rats in the laser only and the laser plus Ad-TTP groups. Two weeks after laser injury, the size of CNV was calculated by perfusion with high-molecular-weight fluorescein isothiocyanate(FITC)-dextran. VEGF m RNA expression in retina-choroid tissue from ten rats in each group was measured by reverse transcription polymerase chain reaction(RT-PCR). RESULTS: Two weeks after treatment, the area of laser-induced CNV was reduced by approximately 60% in the rats given the Ad-TTP injection compared with that in the laser-only group. There was a tendency toward decreased VEGF m RNA expression in the Ad-TTP injection groups.CONCLUSION: A single intravitreal injection of Ad-TTP significantly suppressed CNV size in this experimental laser-induced CNV model. Ad-TTP injection also decreased VEGF m RNA expression compared with that inthe laser-induced CNV group. The present study is meaningful as the first study to investigate the effect of tristetraprolin delivered via intravitreal injection.
文摘AIM:To study the effects of cytokeratin 17(CK17) on sodium iodate(NalO_3) induced rat retinal pigment epithelium(RPE) degeneration,laser induced rat choroidal neovascularization(CNV),and oxidative stress of human retinal pigment epithelium cells(ARPE-19) and human umbilical vein endothelial cell(HUVEC).· METHODS:Thirty 8-week-old male Brown Norway rats were randomly divided into 3 groups,10 rats in control group treated with solvent alone;10 rats in NalO_3 group treated with solvent and 35 mg/kg NalO_3 injection through hypoglossal vein and 10 rats in CK17+NalO_3group treated with 1%CK17 eye drop 3 times a day for1 wk before and 4wk after NalO_3 injection.RPE function was measured with c-wave of electroretinogram(ERG).Another 20 rats were randomly divided into 2 groups.Of them 10 rats in CK17 group were anesthetized to receive Nd:YAG laser and given 1%CK17 eye drop before same as above;10 rats in control were received Nd:YAG and treated with solvent.The development of choroidal neovascularization(CNV) was determined by fundus fluorescein angiography(FFA) performed on 4wk after laser.Methylthiazoly tetrazolium(MTT) assay was used to study effect of CK17 on various oxidants induced injury in ARPE-19 and HUVEC in vitro.· RESULTS:Four weeks after NalO_3 injection,the cwave amplitude of ERG was 0.393±0.02 V in the control group,0.184 ±0.018 V in NalO_3 group and 0.3±0.01 V in CK17+NalO_3 group.There was a significant reversal of the c-wave by CK17 as compared to NalO_3 group(P<0.01).Four weeks after laser,the size of the CNV lesion was2.57±0.27 mm^2 in control group and 1.64±0.08 mm_2 in CK17 group.The lesion size significantly diminished in CK17 group(P<0.01).The in vitro results showed CK17 also reversed the various oxidants induced injuries in ARPE-19 at the dose of 100 μg/mL and enhanced the injury in HUVECs at different concentrations.CONCLUSION:CK17 can significantly protect RPE from NalO_3 induced degeneration in vivo and in vitro and also could reverse the various oxidants induced injuries in vitro.It inhibits the development of CNV in rat model,interfered with vascular endothelial cell proliferation in vitro.
基金Supported by the National Natural Science Foundation of China(No.81760027,No.81860763)Youth Innovation Project of Affiliated Hospital of Inner Mongolia University for Nationalities(No.2018QNJJ01)Young and Middle-aged Ophthalmic Research Fund of Bethune-Lumitin(No.BJ-LM202005)。
文摘AIM:To determine the effects of safranal on choroidal neovascularization(CNV)and oxidative stress damage of human choroidal microvascular endothelial cells(HCVECs)and its possible mechanisms.METHODS:Forty-five rats were used as a laser-induced CNV model for testing the efficacy and safety of safranal(0.5 mg/kg·d,intraperitoneally)on CNV.CNV leakage on fluorescein angiography(FA)and CNV thickness on histology was compared.HCVECs were used for a H_(2)O_(2)-induced oxidative stress model to test the effect of safranal in vitro.MTT essay was carried to test the inhibition rate of safranal on cell viability at different concentrations.Tube formation was used to test protective effect of safranal on angiogenesis at different concentrations.mRNA transcriptome sequencing was performed to find the possible signal pathway.The expressions of different molecules and their phosphorylation level were validated by Western blotting.RESULTS:On FA,the average CNV leakage area was 0.73±0.49 and 0.31±0.11 mm^(2)(P=0.012)in the control and safranal-treated group respectively.The average CNV thickness was 127.4±18.75 and 100.6±17.34μm(P=0.001)in control and safranal-treated group.Under the condition of oxidative stress,cell proliferation was inhibited by safranal and inhibition rates were 7.4%-35.4%at the different concentrations.For tube formation study,the number of new branches was 364 in control group and 35,42,and 17 in 20,40,and 80μg/mL safranal groups respectively(P<0.01).From the KEGG pathway bubble graph,the PI3K-AKT signaling pathway showed a high gene ratio.The protein expression was elevated of insulin receptor substrate(IRS)and the phosphorylation level of PI3K,phosphoinositide-dependent protein kinase 1/2(PDK1/2),AKT and Bcl-2 associated death promoter(BAD)was also elevated under oxidative stress condition but inhibited by safranal.CONCLUSION:Safranal can inhibit CNV both in vivo and in vitro,and the IRS-PI3K-PDK1/2-AKT-BAD signaling pathway is involved in the pathogenesis of CNV.
文摘AIM:To observe the long-term clinical efficacy of intravitreal injections of conbercept,a novel vascular growth factor inhibitor,for the treatment of pathological myopia choroidal neovascularization(PM-CNV).METHODS:A total of 67 eyes(from 67 patients;mean age,54.90±12.7y)with PM-CNV were retrospectively researched.Based on the different schemes used for the administration of the drug,the patients were divided into two groups:group A(n=35;average age,53.31±13.6y;average diopter,9.25±1.72 D),which received only one injection of pro re nata(PRN;1+PRN regimen),and group B(n=32;average age,56.49±11.8y;average diopter,9.63±2.24 D),which received one injection per month for 3mo(3+PRN regimen).Best-corrected visual acuity(BCVA)analysis,intraocular pressure(IOP)examination,slit-lamp microscopy,fundus examination and optical coherence tomography were per formed at each follow-up.The recurrence and treatment times of CNV were recorded.The patients were followed up for at least 12mo.RESULTS:The BCVA was increased in 29 eyes(82.9%)in group A and 30 eyes(93.75%)in group B;no increase or decrease was observed in 6(17.1%)and 2(6.25%)eyes in groups A and B,respectively.The BCVA(log MAR)values before treatment(0.67±0.48 and 0.71±0.56)were significantly higher than those 12mo after treatment(0.31±0.26 and 0.33±0.17)in groups A and B,respectively(P<0.05).The mean central macular thickness(CMT)values had significantly decreased from 346.49±65.99 and 360.10±82.31μm at baseline to 257.29±40.47 and 251.97±48.26μm in groups A and B,respectively,after 12mo of treatment.A total of 21 eyes in group A needed reinjection(60%;average number of injections,2.51±0.98);the corresponding values in group B were 6 eyes(18.75%;average number of injections,3.74±1.22).There were no adverse ocular and systemic complications during the treatment and follow-up.CONCLUSION:Intravitreal injection of conbercept with 1+PRN or 3+PRN improve the visual acuity,reduce macular edema and reduce the level of CMT in patients with PM-CNV.The 3+PRN regimen demonstrates a lower recurrence rate of CNV than the 1+PRN regimen,but requires more treatment.However,both treatment regimens demonstrate long-term safety and efficacy for the treatment of PM-CNV.
基金Supported by National Natural Science Foundation of China(No.81570826)。
文摘AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was treated with celastrol to evaluate this natural compound's impact on CNV.Western blot was applied to explore the possible mechanism for it.For in vitro assay,triplicate for each group was repeated at least three times.For in vivo assay,each group contains 5 mice.RESULTS:Celastrol supressed tube formation and aortic ring sprout neovascularization.In vitro assay exhibited that celastrol inhibiting vascular endothelial growth factor(VEGF)-induced proliferation and migration of human umbilical vein endothelial cells and human choroidal endothelial cells,and by blocking VEGF signaling.Furthermore,intraperitoneal administration of celastrol significantly reduced the area of laser-induced CNV in an in vivo mouse model.By day 14,the area of CNV had decreased by 49.15%and 80.26%in the 0.1 mg/kg celastrol-treated group(n=5)and in the 0.5 mg/kg celastrol treated group(n=5),respectively,compared to the vehicle-treated group(n=5).CONCLUSION:Celastrol inhibits CNV by inhibiting VEGF-induced proliferation and migration of vascular endothelial cells,indicating that celastrol is a potent,natural therapeutic compound for the prevention of CNV.
基金Supported by the Swiss RetinAward 2017 from the Swiss Vitreo Retinal Group(SVRG)Bayer AG+2 种基金CSC(Chinese Scholarship Council)EAKAS(Swiss Excellence Scholarship)Natural Science Basic Research Program of Shaanxi,China(No.2020JM-400)。
文摘AIM:To compare choroidal neovascularization(CNV)lesion measurements obtained by in vivo imaging modalities,with whole mount histological preparations stained with isolectin GS-IB4,using a murine laser-induced CNV model.METHODS:B6 N.Cg-Tg(Csf1 r-EGFP)1 Hume/J heterozygous adult mice were subjected to laser-induced CNV and were monitored by fluorescein angiography(FA),multicolor(MC)fundus imaging and optical coherence tomography angiography(OCTA)at day 14 after CNV induction.Choroidalretinal pigment epithelium(RPE)whole mounts were prepared at the end of the experiment and were stained with isolectin GS-IB4.CNV areas were measured in all different imaging modalities at day 14 after CNV from three independent raters and were compared to choroidal-RPE whole mounts.Intraclass correlation coefficient(ICC)type 2(2-way random model)and its 95%confidence intervals(CI)were calculated to measure the correlation between different raters’measurements.Spearman’s rank correlation coefficient(Spearman’s r)was calculated for the comparison between FA,MC and OCTA data and histology data.RESULTS:FA(early and late)and MC correlates well with the CNV measurements ex vivo with FA having slightly better correlation than MC(FA early Spearman’s r=0.7642,FA late Spearman’s r=0.7097,and MC Spearman’s r=0.7418),while the interobser ver reliability was good for both techniques(FA early ICC=0.976,FA late ICC=0.964,and MC ICC=0.846).In contrast,OCTA showed a poor correlation with ex vivo measurements(Spearman’s r=0.05716)and high variability between different raters(ICC=0.603).CONCLUSION:This study suggests that FA and MC imaging could be used for the evaluation of CNV areas in vivo while caution must be taken and comparison studies should be performed when OCTA is employed as a CNV monitoring tool in small rodents.
基金Supported by Medical and Health Platform Project of Zhejiang Province(No.2021KY810)。
文摘AIM:To analyse macular microvascular alterations in myopic choroidal neovascularization(m CNV)and the efficiency of anti-vascular endothelial growth factor(antiVEGF)therapy for m CNV by optical coherence tomography angiography(OCTA).METHODS:A total of 123 patients were included in this retrospective study,divided into m CNV group,high myopia(HM)group,and normal group at the Affiliated Eye Hospital of Wenzhou Medical University from January 2017 to January 2019.Superficial vessel density,deep capillary density,foveal avascular zone(FAZ)area,A-circularity index(AI)and vessel density around the 300μm width of the FAZ region density(FD)and the area of choroidal neovascularization(CNV)lesion(only for m CNV group)were measured on 3×3 mm2 OCTA images.FAZ area was corrected for axial length.Central macular thickness(CMT)was measured on OCT in m CNV group.Compared the parameters on OCTA of 3 groups and pre-anti-VEGF and post-anti-VEGF at 1,2,3,and 6 mo follow-up in m CNV group.RESULTS:There were significant differences among 3 groups in superficial vessel density,deep capillary density and FD(P<0.05).FAZ area in HM group was smaller than normal group(P<0.05),but there was no significant difference between m CNV group and the other two group.AI increased in m CNV group(P<0.05).The mean CMT,area and flow area of CNV lesion decreased after treatment(P<0.05),while vessel density and FAZ didn’t change.The mean CMT,area and flow area of CNV lesion statistically decreased after anti-VEGF treatment in m CNV group(P<0.05),while superficial vessel density,deep capillary density and FAZ area,AI and FD didn’t change.The mean reduction ratio of lesions was 50.32%(7.07%to 100%).Lesion regression 100%was observed in 2 cases(4.88%).There was a negative correlation between the CNV lesion area and reduction ratio(r=-0.380,P=0.042)and the flow lesion area and reduction ratio(r=-0.402,P=0.030).CONCLUSION:Macular vessel density decreases,FAZ turns smaller and more irregular in m CNV eyes.AntiVEGF therapy is efficient for m CNV without affecting vessel density and FAZ,but it is unable to completely eliminate CNV lesions in most cases.The bigger m CNV lesions have lower reduction ratio.
文摘Dear Editor,I am Cheolmin Yun,from the Department of Ophthalmology,Korea University College of Medicine.I write to present a case report of a female patient with a myopic patient suffering from atrophic choroidal neovascularization(CNV)and a full thickness macular hole(FTMH),who was treated with an intravitreal
基金Supported by the Shaanxi Key Research and Development Program-General Project in the Field of Social Development (No.2017SF-140)。
文摘AIM:To investigate whether anti-placental growth factor(PGF) can inhibit subretinal fibrosis and whether this effect is mediated by the inhibitory effect of PGF on epithelial-mesenchymal transition(EMT) of retinal pigment epithelial(RPE) cells.METHODS:Subretinal fibrosis model was established in laser induced choroidal neovascularization(CNV) mice on day 21 after laser photocoagulation.Immunofluorescence staining(IFS) of cryosections and enzyme-linked immunosorbent assay(ELISA) were used to detect the expression of PGF.IFS of whole choroidal flat-mounts was used to detect the degree of subretinal fibrosis.IFS of cryosections and ELISA were used to detect the expression of EMT related indicators in subretinal fibrosis lesions.RESULTS:The expression of PGF protein in subretinal fibrosis lesions was significantly up-regulated(P<0.05),and mainly co-stained with pan-cytokeratin labeled RPE cells.Intravitreal injection of anti-PGF neutralizing antibody reduced the area of subretinal fibrosis and the ratio of fibrotic/angiogenic area significantly at the concentrations of 0.25,0.5,1.0,and 2.0 μg/μL(all P<0.05).The expression of E-cadherin in the local RPE cells decreased,while α-SMA increased significantly in subretinal fibrosis lesions,and the application of anti-PGF neutralizing antibody could reverse these changes(P<0.05).CONCLUSION:The expression of PGF is up-regulated in the lesion site of subretinal fibrosis and mainly expressed in RPE cells.Intravitreal injection of anti-PGF neutralizing antibody can significantly inhibit the degree of subretinal fibrosis in CNV mice,and this effect may be mediated by the inhibition of PGF on EMT of RPE cells.
基金Supported by National Key R&D Program of China(No.2020YFC2008200)Capital Clinical Diagnosis and Treatment Technology Research and Demonstration Application Project of China(No.Z191100006619029)。
文摘AIM: To investigate the clinical characteristics and genetic features of a Bietti crystalline dystrophy(BCD) proband in a Chinese family.METHODS: A Chinese female diagnosed with BCD complicated by bilateral choroidal neovascularization(CNV) and her parents underwent complete ophthalmic examinations, including fundus autofluorescence(AF), fundus photography(FP), fundus fluorescein angiography(FFA), visual field testing, full-field electroretinography(ERG), optical coherence tomography(OCT) and optical coherence tomography angiography(OCTA). The sequencing of the CYP4 V2 gene was performed to the whole family.RESULTS: Bilateral tiny glittering crystal-like deposits and differing extent of atrophy of the retinal pigment epithelium(RPE) were found in the posterior pole of her fundus. The diffuse hypo-fluorescence shown on AF images and window defects shown on FFA both indicated the atrophy of the RPE and choriocapillaris. OCT showed the thinning of the RPE and choriocapillaris layer, ellipsoid zone(EZ) band defect and CNV in both eyes. OCTA images proofed bilateral type 2 CNV. The visual field test showed central and paracentral scotoma. ERG showed a slightly decreased b-wave in scotopic ERG. Gene sequencing identified three mutations of the CYP4 V2 gene, c.802_807 del, c.810 del T, and c.1388 G>A. The mutation c.1388 G>A was a novel substitution mutation.CONCLUSION: The novel mutation c.1388 G>A may be a possible cause that could induce the clinical phenotype of BCD.
基金the Army Laboratory Animal Foundation of China(No.SYDW2016-011)。
文摘AIM:To investigate therapeutic effects of traditional Chinese medicine formulations,Hexuemingmu(HXMM)on laser-induced choroidal neovascularization(CNV)and followup effect in mice.METHODS:C57 BL/6 mice of 8-week-old were used and CNV was induced with 577 nm laser photocoagulation.Animals were randomly divided into groups and different doses of HXMM were administered daily.One,four,and eight weeks after the intervention,the electroretinogram(ERG),fundus fluorescence angiography,choroidal flat mount and immunofluorescence staining were preformed to evaluate the function and CNV formation.The expression levels of angiogenic proteins were determined by Western blotting and immunofluorescence staining.An analysis of variance and Kruskal-Wallis test were used to test the differences among the groups.RESULTS:The results showed that HXMM effectively increased amplitude of ERG of mice(P<0.05),alleviated fundus CNV leakage(P<0.05),and reduced the area of neovascularization and the expression of angiogenic proteins(P<0.05)after laser-induced CNV.CONCLUSION:HXMM can protect the retinal function of mice after laser-induced CNV,and inhibit the CNV development.
文摘Dear Editor,My name is Georgios Tsokolas and I am currently working as Medical Retina Research Fellow at the Eye Unit of Southampton General Hospital in United Kingdom.I am writing this letter to present three case series of choroidal neovascular(CNV)membrane after macular surgery。
