Background Atresia and degeneration,a follicular developmental fate that reduces female fertility and is triggered by granulosa cell(GC)apoptosis,have been induced by dozens of miRNAs.Here,we report a miRNA,miR-423,th...Background Atresia and degeneration,a follicular developmental fate that reduces female fertility and is triggered by granulosa cell(GC)apoptosis,have been induced by dozens of miRNAs.Here,we report a miRNA,miR-423,that inhibits the initiation of follicular atresia(FA),and early apoptosis of GCs.Results We showed that miR-423 was down-regulated during sow FA,and its levels in follicles were negatively correlated with the GC density and the P4/E2 ratio in the follicular fluid in vivo.The in vitro gain-of-function experiments revealed that miR-423 suppresses cell apoptosis,especially early apoptosis in GCs.Mechanically speaking,the miR-423 targets and interacts with the 3’-UTR of the porcine SMAD7 gene,which encodes an apoptosis-inducing factor in GCs,and represses its expression and pro-apoptotic function.Interestingly,FA and the GC apoptosis-related lncRNA NORHA was demonstrated as a ceRNA of miR-423.Additionally,we showed that a single base deletion/insertion in the miR-423 promoter is significantly associated with the number of stillbirths(NSB)trait of sows.Conclusion These results demonstrate that miR-423 is a small molecule for inhibiting FA initiation and GC early apoptosis,suggesting that treating with miR-423 may be a novel approach for inhibiting FA initiation and improving female fertility.展开更多
Background:Dihydroartemisinin(DHA)is reported to be a potential anticancer agent,and the mechanisms underlying the effects of DHA on diffuse large B cell lymphoma however are still obscure.This study aimed to assess t...Background:Dihydroartemisinin(DHA)is reported to be a potential anticancer agent,and the mechanisms underlying the effects of DHA on diffuse large B cell lymphoma however are still obscure.This study aimed to assess the antitumor effect of DHA on diffuse large B cell lymphoma cells and to determine the potential underlying mechanisms of DHA-induced cell apoptosis.Methods:Here,the Cell Counting Kit 8 assay was conducted to study cell proliferation.We performed Annexin V-FITC/propidium iodide staining,real-time polymerase chain reaction,and western blot analysis to analyze cell apoptosis and potential molecular mechanisms.Results:The results showed that DHA substantially suppressed cell proliferation and induced cell apoptosis in vitro in a time-and concentration-dependent fashion.Moreover,STAT3 activity could be inhibited after stimulation with DHA.Conclusion:These results imply that the underlying anti-tumoral effect of DHA may increase apoptosis in diffuse large B cell lymphoma cells via the STAT3 signaling pathway.In addition,DHA might be an effective drug for diffuse large B cell lymphoma therapy.展开更多
AIM:To determine whether an antisense RNA corresponding to the human Alu transposable element(Aluas RNA)can protect human lens epithelial cells(HLECs)from methylglyoxal-induced apoptosis.METHODS:Cell counting kit-8(CC...AIM:To determine whether an antisense RNA corresponding to the human Alu transposable element(Aluas RNA)can protect human lens epithelial cells(HLECs)from methylglyoxal-induced apoptosis.METHODS:Cell counting kit-8(CCK-8)and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assays were used to assess HLEC viability.HLEC viability/death was detected using a Calcein-AM/PI double staining kit;the annexin V-FITC method was used to detect HLEC apoptosis.The cytosolic reactive oxygen species(ROS)levels in HLECs were determined using a reactive species assay kit.The levels of malondialdehyde(MDA)and the antioxidant activities of total-superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px)were assessed in HLECs using their respective kits.RT-q PCR and Western blotting were used to measure m RNA and protein expression levels of the genes.RESULTS:Aluas RNA rescued methylglyoxal-induced apoptosis in HLECs and ameliorated both the methylglyoxalinduced decrease in Bcl-2 m RNA and the methylglyoxalinduced increase in Bax m RNA.In addition,Aluas RNA inhibited the methylglyoxal-induced increase in Alu sense RNA expression.Aluas RNA inhibited the production of ROS induced by methylglyoxal,restored T-SOD and GSHPx activity,and moderated the increase in MDA content after treatment with methylglyoxal.Aluas RNA significantly restored the methylglyoxal-induced down-regulation of Nrf2 gene and antioxidant defense genes,including glutathione peroxidase,heme oxygenase 1,γ-glutamylcysteine synthetase and quinone oxidoreductase 1.Aluas RNA ameliorated methylglyoxal-induced increases of the m RNA and protein expression of Keap1 that is the negative regulator of Nrf2.CONCLUSION:Aluas RNA reduces apoptosis induced by methylglyoxal by enhancing antioxidant defense.展开更多
White tea encompasses a number of teas unique to eastern Fujian in China. Although white tea extracts have been reported to result in cancer cell apoptosis, to date, few studies have evaluated the mechanism of such ap...White tea encompasses a number of teas unique to eastern Fujian in China. Although white tea extracts have been reported to result in cancer cell apoptosis, to date, few studies have evaluated the mechanism of such apoptotic induction. A transcription factor that plays a critical role in cell apoptosis, NF-κB p65, is also likely critical by which white tea extracts induce cancer cell apoptosis. In this study, white tea aqueous extract (WTAE) was added to BEL-7402 and Hela cell media and NF-κB p65 activation was evaluated using western blotting and immunofluorescence. Results revealed that the phosphorylation of IKBα and p65 decreased in both cell lines after WTAE treatment. And the nuclear translocation of NF-κB p65 in both cell lines was also reduced with the WTAE treatment. NF-κB p65 inhibition was noted to accelerate apoptosis. Our findings suggest that NF-κB p65 was an important modulator in WTAE-induced apoptotic signal transduction and it acted as a negative regulator of apoptotic induction in BEL-7402 and Hela cancer cell lines.展开更多
The retinal ganglion cells of the optic nerve have a limited capacity for self-repair after injury.Valproate is a histone deacetylase inhibitor and multitarget drug,which has been demonstrated to protect retinal neuro...The retinal ganglion cells of the optic nerve have a limited capacity for self-repair after injury.Valproate is a histone deacetylase inhibitor and multitarget drug,which has been demonstrated to protect retinal neurons.In this study,we established rat models of optic nerve-crush injury and injected valproate into the vitreous cavity immediately after modeling.We evaluated changes in the ultrastructure morphology of the endoplasmic reticulum of retinal ganglion cells over time via transmission electron microscope.Immunohistochemistry and western blot assay revealed that valproate upregulated the expression of the endoplasmic reticulum stress marker glucose-regulated protein 78 and downregulated the expression of transcription factor C/EBP homologous protein,phosphorylated eukaryotic translation initiation factor 2α,and caspase-12 in the endoplasmic reticulum of retinal ganglion cells.These findings suggest that valproate reduces apoptosis of retinal ganglion cells in the rat after optic nerve-crush injury by attenuating phosphorylated eukaryotic translation initiation factor 2α-C/EBP homologous protein signaling and caspase-12 activation during endoplasmic reticulum stress.These findings represent a newly discovered mechanism that regulates how valproate protects neurons.展开更多
BACKGROUND As an active ingredient derived from Dioscorea zingiberensis C.H.Wright,deltonin has been reported to show anti-cancer effects in a variety of malignancies.AIM To investigate the role and mechanism of actio...BACKGROUND As an active ingredient derived from Dioscorea zingiberensis C.H.Wright,deltonin has been reported to show anti-cancer effects in a variety of malignancies.AIM To investigate the role and mechanism of action of deltonin in promoting gastric carcinoma(GC)cell apoptosis and chemosensitivity to cisplatin.METHODS The GC cell lines AGS,HGC-27,and MKN-45 were treated with deltonin and then subjected to flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-3,5-diphenyltet-razolium bromide assays for cell apoptosis and viability determination.Western blot analysis was conducted to examine alterations in the expression of apoptosis-related proteins(Bax,Bid,Bad,and Fas),DNA repair-associated proteins(Rad51 and MDM2),and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of the rapamycin(PI3K/AKT/mTOR)and p38-mitogen-activated protein kinase(MAPK)axis proteins.Additionally,the influence of deltonin on GC cell chemosensitivity to cisplatin was evaluated both in vitro and in vivo.RESULTS Deltonin treatment weakened viability,enhanced apoptosis,and dampened DNA repair in GC cell lines in a dose-dependent pattern.Furthermore,deltonin mitigated PI3K,AKT,mTOR,and p38-MAPK phosphorylation.HS-173,an inhibitor of PI3K,attenuated GC cell viability and abolished deltonin inhibition of GC cell viability and PI3K/AKT/mTOR and p38-MAPK pathway activation.Deltonin also promoted the chemosensitivity of GC cells to cisplatin via repressing GC cell proliferation and growth and accelerating apoptosis.CONCLUSION Deltonin can boost the chemosensitivity of GC cells to cisplatin via inactivating p38-MAPK and PI3K/AKT/mTOR signaling.展开更多
Marine collagen peptides(MCPs)are natural products prepared by hydrolyzing marine collagen protein through a variety of chemical methods or enzymes.MCPs have a range of structures and biological activities and are wid...Marine collagen peptides(MCPs)are natural products prepared by hydrolyzing marine collagen protein through a variety of chemical methods or enzymes.MCPs have a range of structures and biological activities and are widely present in marine species.MCPs also have a small molecular weight,are easily modified,and absorbed by the body.These properties have attracted great interest from researchers studying antioxidant,anti-tumor,and anti-aging activities.MCPs of specific molecular weights have significant anti-tumor activity and no toxic side effects.Thus,MCPs have the potential use as anti-cancer adjuvant drugs.Free radicals produced by oxidation are closely related to human aging,cancer,arteriosclerosis,and other diseases,but their relationship with cancer is not well known.In this review,we focus on the antioxidant properties of MCPs in the treatment of cancer,highlighting their antioxidant molecular structure and potential for clinical practice.展开更多
AIM:To investigate the effect of electroacupuncture(EA)on the mitochondria-dependent apoptotic signaling pathway in the ciliary muscle of guinea pigs with negative lensinduced myopia(LIM).METHODS:Guinea pigs were rand...AIM:To investigate the effect of electroacupuncture(EA)on the mitochondria-dependent apoptotic signaling pathway in the ciliary muscle of guinea pigs with negative lensinduced myopia(LIM).METHODS:Guinea pigs were randomly divided into normal control(NC)group,LIM group,LIM+SHAM acupoint(LIM+SHAM)group,and LIM+EA group.Animals in the NC group received no intervention,while those in other three groups were covered with-6.0 diopter(D)lenses on right eyes.Meanwhile,animals in the LIM+EA group received EA at Hegu(LI4)combined with Taiyang(EX-HN5)acupoints,while those in the LIM+SHAM group were treated at sham points.After treatments for 1,2,and 4wk,morphological changes in ciliary muscles were observed with hematoxylin and eosin(H&E)staining and nick end labeling(TUNEL),and the expression of the mitochondrial apoptotic signaling pathway-related molecules in ciliary muscles was measured by real-time quantitative polymerase chain reaction(qPCR)and Western blot.Additionally,the adenosine triphosphate(ATP)contents were also determined in ciliary muscles.RESULTS:Axial length increased significantly in the LIM and LIM+SHAM groups and decreased in the LIM+EA group.The ciliary muscle fibers were broken and destroyed in both LIM and LIM+SHAM groups,whereas those in the LIM+EA group improved significantly.TUNEL assay showed the number of apoptotic cells increased in the LIM and LIM+SHAM groups,whereas reduced in the LIM+EA group.ATP contents showed a significant decrease in the LIM and LIM+SHAM groups,whereas increased after EA treatment.Compared with the NC group,the dynamin-related protein 1(DRP1),Caspase3,and apoptotic protease activator 1(APAF1)levels were significantly increased in the LIM group and decreased in the LIM+EA group.CONCLUSION:The results provide evidence of EA inhibiting the development of myopia by regulating the mitochondrial apoptotic signaling pathway.展开更多
BACKGROUND Gastric adenocarcinoma(GAC)mortality rates have remained relatively changed over the past 30 years,and it continues to be one of the leading causes of cancerrelated death.AIM To search for novel miRNAs rela...BACKGROUND Gastric adenocarcinoma(GAC)mortality rates have remained relatively changed over the past 30 years,and it continues to be one of the leading causes of cancerrelated death.AIM To search for novel miRNAs related to GAC prognosis and further investigate the effect of miR-96-5p on MGC-803 cells.METHODS The miRNA expression profile data of GAC based on The Cancer Genome Atlas were obtained and used to screen differently expressed miRNAs(DEMs)and DEMs related to GAC prognosis.Then,the expression of DEMs related to GAC prognosis was identified in GAC tumor samples and adjacent normal samples by qRT-PCR.The target gene,ZDHHC5,of miR-96-5p was predicted using TargetScan,miRTarBase,and miRDB databases and confirmed by luciferase reporter assay.Furthermore,MGC-803 cells were transfected with inhibitor NC,miR-96-5p inhibitor,si-ZDHHC5,or miR-96-5p inhibitor+si-ZDHHC5,and then cell apoptosis was detected by flow cytometry.The expression of ZDHHC5,Bcl-2,and COX-2 was detected using western blotting.RESULTS A total of 299 DEMs and 35 DEMs related to GAC prognosis were screened based on The Cancer Genome Atlas.Then compared with adjacent normal samples,the levels of miR-96-5p,miR-222-5p,and miR-652-5p were remarkably increased,while miR-125-5p,miR-145-3p,and miR-379-3p levels were reduced in GAC tumor samples(P<0.01),which were consistent with bioinformatics analysis.Furthermore,ZDHHC5 was defined as a direct target gene of miR-96-5p.miR-96-5p inhibition increased the number of apoptotic cells as well as promoted the expression of ZDHHC5,Bcl-2,and COX-2 in MGC-803 cells(P<0.01).After ZDHHC5 inhibition,the number of apoptotic cells and the expression of ZDHHC5,Bcl-2,and COX-2 were reduced.The addition of an miR-96-5p inhibitor partly reversed these effects(P<0.01).CONCLUSION Our findings identified six miRNAs related to GAC prognosis and suggested that downregulated miR-96-5p might induce cell apoptosis via upregulating ZDHHC5 expression in MGC-803 cells.展开更多
BACKGROUND: Neuronal apoptosis in perihematomal brain tissues following intracerebral hemorrhage is strongly related to the formation of a compound signal pathway between nerve growth factor precursor (proNGF), p75NTR...BACKGROUND: Neuronal apoptosis in perihematomal brain tissues following intracerebral hemorrhage is strongly related to the formation of a compound signal pathway between nerve growth factor precursor (proNGF), p75NTR, and sortilin receptor. Sortilin acts as a co-receptor and molecular switch governing the p75NTR-mediated pro-apoptotic signal induced by proNGF. OBJECTIVE: To investigate proNGF and sortilin expressions in perihematomal brain tissues following intracerebral hemorrhage, and to study the effects of proNGF and sortilin on secondary cell apoptosis. DESIGN, TIME AND SETTING: A paired, comparison study was performed at the Laboratory of Histology and Embryology, Xi'an Jiaotong University from October 2007 to September 2008. MATERIALS: Brain tissue samples were obtained from 15 patients with intracerebral hemorrhage, who were treated at the Department of Neurosurgery, First Affiliated Hospital, Medical College of Xi'an Jiaotong University from October 2007 to March 2008. Rabbit anti-proNGF polyclonal antibody was provided by Chemicon, USA; rabbit antisortilin polyclonal antibody by Abcam, UK; and TUNEL kit by Promega, USA. METHODS: Perihematomal brain tissues selected 0.5 cm from the hemorrhage area were considered to be the hemorrhage group, while brain tissues from the middle temporal gyrus served as the control group. MAIN OUTCOME MEASURES: Histopathological changes were detected using hematoxylin-eosin staining, cell apoptosis was determined using the TUNEL method, and proNGF and sortilin ex- pressions were determined using immunohistochemistry. RESULTS: Edema was clearly observed in perihematomal brain tissues, and infiltration of inflammatory cells was visible, with the presence of irregular and necrotic bodies. The apoptotic rate in the hemorrhage group was significantly greater than in the control group (P < 0.01). Moreover, sortilin expression significantly increased (P < 0.01), but proNGF expression remained unchanged (P > 0.05). Correlation analysis suggested that sortilin expression positively correlated with apoptosis (rs = 0.648, P < 0.01). CONCLUSION: proNGF expression was stable, but sortilin expression increased in perihematomal brain tissues following intracerebral hemorrhage, suggesting that sortilin acted as a co-receptor and molecular switch to govern p75NTR-mediated cell apoptosis.展开更多
Background:Follicular atresia has been shown to be strongly associated with a low follicle utilization rate and female infertility,which are regulated by many factors such as microRNAs(miRNAs),which constitute a class...Background:Follicular atresia has been shown to be strongly associated with a low follicle utilization rate and female infertility,which are regulated by many factors such as microRNAs(miRNAs),which constitute a class of noncoding RNAs(ncRNAs).However,little is known about long noncoding RNAs(lncRNAs),which constitute another ncRNA family that regulate follicular atresia.Results:A total of 77 differentially expressed lncRNAs,including 67 upregulated and 10 downregulated lncRNAs,were identified in early atretic follicles compared to healthy follicles by RNA-Sequencing.We characterized a noncoding RNA that was highly expressed in atretic follicles(NORHA).As an intergenic lncRNA,NORHA was one of the upregulated lncRNAs identified in the atretic follicles.To determine NORHA function,RT-PCR,flow cytometry and western blotting were performed,and the results showed that NORHA was involved in follicular atresia by influencing GC apoptosis with or without oxidative stress.To determine the mechanism of action,bioinformatics analysis,luciferase reporter assay and RNA immunoprecipitation assay were performed,and the results showed that NORHA acted as a‘sponge’,that directly bound to the miR-183-96-182 cluster,and thus prevented its targeted inhibition of FoxO1,a major sensor and effector of oxidative stress.Conclusions:We provide a comprehensive perspective of lncRNA regulation of follicular atresia,and demonstrate that NORHA,a novel lncRNA related to follicular atresia,induces GC apoptosis by influencing the activities of the miR-183-96-182 cluster and FoxO1 axis.展开更多
Objective Epidemiological studies reveal that exposure to fine particulate matter(aerodynamic diameter≤2.5μm,PM_(2.5))increases the morbidity and mortality of respiratory diseases.Emerging evidence suggests that hum...Objective Epidemiological studies reveal that exposure to fine particulate matter(aerodynamic diameter≤2.5μm,PM_(2.5))increases the morbidity and mortality of respiratory diseases.Emerging evidence suggests that human circulating extracellular vesicles(EVs)may offer protective effects against injury caused by particulate matter.Currently,however,whether EVs attenuate PM_(2.5)-induced A549 cell apoptosis is unknown.Methods EVs were isolated from the serum of healthy subjects,quantified via nanoparticle tracking analysis,and qualified by the marker protein CD63.PM_(2.5)-exposed(50μg/mL)A549 cells were pretreated with 10μg/mL EVs for 24 h.Cell viability,cell apoptosis,and AKT activation were assessed via Cell Counting Kit-8,flow cytometry,and Western blot,respectively.A rescue experiment was also performed using MK2206,an AKT inhibitor.Results PM_(2.5)exposure caused a 100%in crease in cell apoptosis.EVs treatme nt reduced cell apoptosis by 10%,promoted cell survival,and inhibited the PM_(2.5)-induced upregulation of Bax/Bcl2 and cleaved caspase 3/caspase 3 in PM_(2.5)-exposed A549 cells.Moreover,EVs treatment reversed PM_(2.5)-induced reductions in p-AKT^(Thr308)and p-AKT^(Ser473).A KT inhibition attenuated the anti-apoptotic effect of EVs treatment on PM_(2.5)-exposed A549 cells.Conclusions EVs treatment promotes cell survival and attenuates PM_(2.5)-induced cell apoptosis via AKT phosphorylation.Human serum-derived EVs may be an efficacious novel therapeutic strategy in PM_(2.5)-induced lung injury.展开更多
BACKGROUND:Many Chinese herbs,especially herbal injections,have been shown to have anti-tumor effects in recent years.However,since most reports focus on the clinical effectiveness of these herbs,their mechanisms of a...BACKGROUND:Many Chinese herbs,especially herbal injections,have been shown to have anti-tumor effects in recent years.However,since most reports focus on the clinical effectiveness of these herbs,their mechanisms of action are not well understood.In this study,we assessed apoptosis in the hepatocellular carcinoma (HCC) cell line HepG2 induced by an injectable extract from the seed of Coix lacryma-jobi (Semen coicis,SC),and monitored the expression of Bcl-2 and caspase-8.METHODS:Injectable SC was applied to HepG2 cells at different concentrations and the cells were collected 12,24 and 48 hours later.5-fluorouracil was used as a positive control group,and fluorescence-activated cell-sorting cytometry was used to measure the apoptosis rate of HepG2 cells and the expression of Bcl-2 and caspase-8 proteins.RESULTS:SC induced apoptosis in HepG2 cells in a concentration and time-dependent manner,and the expression of caspase-8 was elevated and prolonged.However,it did not significantly influence the expression of Bcl-2.CONCLUSION:Injectable SC may induce apoptosis in HCC cells by regulating the expression of caspase-8.展开更多
Objective:To study the protective effect of resveratrol on lens epithelial cell apoptosis in diabetic cataract rat.Methods:A total of 84 Wistar rats were divided into 4 groups:12 in Group A(control group).24 in Group ...Objective:To study the protective effect of resveratrol on lens epithelial cell apoptosis in diabetic cataract rat.Methods:A total of 84 Wistar rats were divided into 4 groups:12 in Group A(control group).24 in Group B(diabetic cataract group),24 in Group C(therapeutic-dose of resveratrol group) and 24 in Group D(low-dose of resveratrol group).Rats in Group B-D were given with60 mg/kg streptozotocin through intraperitoneal injection.Rats in Group C were given with 100mg/kg resveratrol and rats in Group D were given with 20 mg/kg resveratrol.The caspase-3expression levels and apoptosis ratios of LEC among each group were observed:the degrees of lens opacity in Group B-D after 12 weeks were compared.Results:There were significant differences in caspase-3 expression levels,apoptosis ratios of T.F.C among groups at 4 w,8 w and12 w(P<0.05).After 12 weeks,in Group B the degree of lens opacity was as follow:0(0.00%) in grade Ⅰ,3(37.50%) in grade Ⅱ,2(25.00%)in grade Ⅲ,2(25.00%)grade Ⅳ,and 1(12.50%) in gradeⅤ:in Group C:2(25.00%)in grade Ⅰ,4(50.00%) in grade Ⅱ.2(25.00%)in grade Ⅲ,0(0.00%)gradeⅣ,and 0(0.00%) in grade Ⅴ;in Group D:1(12.50%)in grade Ⅰ,4(50.00%) in grade Ⅱ,2(25.00%)in grade Ⅲ,1(12.50%) grade Ⅳ,and 0(0.00%) in grade Ⅴ.The.difference among Group B-D was statistically significant(P<0.05).Conclusions:Resveratrol has protective effect on lens epithelial cell apoptosis in diabetic cataract rat,and the effect is relative to its dose.展开更多
Apoptosis contributes to the development of diabetic nephropathy, but the mechanism by which high glucose induces apoptosis is not fully understood. Apoptosis of tubular epithelial cells is a major feature of diabetic...Apoptosis contributes to the development of diabetic nephropathy, but the mechanism by which high glucose induces apoptosis is not fully understood. Apoptosis of tubular epithelial cells is a major feature of diabetic kidney disease, and hyperglycemia triggers the generation of free radicals and oxidant stress in tubular cells. Hyperglycemia and high glucose in vitro also lead to apoptosis, a form of programmed cell death. High glucose similar to those seen with hyperglycemia in people with diabetes mellitus, lead to accelerated apoptosis, a form of programmed cell death characterized by cell shrinkage, chromatin condensation and DNA fragmentation, in variety of cell types, including renal proximal tubular epithelial cells.展开更多
AIM:To investigate whether Melatonin has synergistic effects with Doxorubicin in the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and Bel-7402.METHODS:The synergism of Melatonin and Dox...AIM:To investigate whether Melatonin has synergistic effects with Doxorubicin in the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and Bel-7402.METHODS:The synergism of Melatonin and Doxorubicin inhibited the cell growth and induced cell apoptosis in human hepatoma cell lines HepG2 and Bel-7402.Cell viability was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT)assay.Cell apoptosis was evaluated using TUNEL method and flow cytometry.Apoptosis-related protein Bax,Bcl-2 and caspase-3 expressions were measured by immunohistochemical staining.RESULTS:Treatment with Melatonin(10 -8 -10 -5 mol/L) alone had a dose-related inhibitory effect on cell proliferation but no cytotoxic effect on hepatoma cell lines HepG2 and Bel-7402.Interestingly,when combined with Doxorubicin,Melatonin significantly increased the effects of cell growth inhibition and cell apoptosis.Furthermore,TUNEL staining and flow cytometry revealed that cooperative apoptosis induction was associated with decreased expression of Bcl-2 as well as increased expression of Bax and Caspase3.CONCLUSION:The synergism of Melatonin and Doxorubicin inhibits hepatoma cell growth and induces cell apoptosis.展开更多
Background: Heat stress is known to alter follicular dynamics and granulosa cell function and may contribute to the diminished reproductive efficiency commonly observed in mammals during the summer. Although several i...Background: Heat stress is known to alter follicular dynamics and granulosa cell function and may contribute to the diminished reproductive efficiency commonly observed in mammals during the summer. Although several investigators have studied heat-induced ovarian injury, few reports have focused on the effects of chronic heat stress on ovarian function and the molecular mechanisms through which it induces ovarian injury.Methods: In Exp. 1, 48 female mice were assigned to a control or heat-stressed treatment. After exposure to a constant temperature of 25 °C for 7, 14, 21 or 28 d(n = 6) or to 42 °C for 3 h per d for 7, 14, 21 or 28 d(n = 6), the mice were euthanized and their ovaries were analyzed for follicular atresia, granulosa cell apoptosis, changes in the abundance of HSP70 protein and serum concentrations of estradiol. In Exp. 2, the expression of HSP70 and aromatase was quantified in antral follicles cultured in vitro at 37 or 42 °C for 24 h. In Exp. 3, granulosa cells from ovaries maintained at 37 or 41 °C for 2 h were analyzed for their expression of HSP70, Bim, caspase-3 and cleaved caspase-3.Results: In Exp. 1, body weight and food intake of heat-stressed mice decreased(P < 0.05) compared with control mice while the concentration of estradiol in serum was lower(P < 0.05) in heat-stressed mice than in control mice. Compared with control mice, the percentage of atretic follicles and the number of antral follicles with severe apoptotic signals were increased(P < 0.05) after 21 d of heat-stressed treatment. HSP70 protein was more abundant(P < 0.05) in heat-stressed mice than control mice. In Exp. 2, heat stress increased HSP70 and decreased aromatase proteins(P < 0.05) in antral follicles. In Exp. 3, TUNEL-positive granulosa cells from heat-stressed ovaries were observed concomitant with a significant increase in HSP70, Bim and cleaved caspase-3 protein.Conclusion: Heat-stress in mice decrease estradiol in serum and aromatase in antral follicles but increased number of atretic follicles and granulosa cell undergoing apoptosis which may explain the decreased fertility commonly observed in heat-stressed animals.展开更多
AIM: To investigate the apoptotic effects of melittin on SGC-7901 cells via activation of the mitochondrial signaling pathway in vitro.METHODS: SGC-7901 cells were stimulated by melittin, and its effect on proliferati...AIM: To investigate the apoptotic effects of melittin on SGC-7901 cells via activation of the mitochondrial signaling pathway in vitro.METHODS: SGC-7901 cells were stimulated by melittin, and its effect on proliferation and apoptosis of was investigated by methyl thiazolyl tetrazolium assay, morphologic structure with transmission electron microscopy, annexin-V/propidium iodide double-staining assay, measuring mitochondrial membrane potential(MMP) levels, and analyzing reactive oxygen species(ROS) concentrations were analyzed by flow cytometry. Cytochrome C(Cyt C), apoptosis-inducing factor(AIF), endonuclease G(Endo G), second mitochondria-derived activator of caspases(Smac)/direct IAP binding protein with low isoelectric point(Diablo), and FAS were analyzed by western blot. The expression of caspase-3 and caspase-8 was measured using activity assay kits.RESULTS: Melittin was incubated at 1.0, 2.0, 4.0, or 6.0 μg/m L for 1, 2, 4, 6, or 8 h and showed a timeand concentration-dependent inhibition of SGC-7901 cell growth. Melittin induced SGC-7901 cell apoptosis, which was confirmed by typical morphological changes. Treatment with 4 μg/m L melittin induced early apoptosis of SGC-7901 cells, and the early apoptosis rates were 39.97% ± 3.19%, 59.27% ± 3.94%, and 71.50% ± 2.87% vs 32.63% ± 2.75% for 1, 2, and 4 h vs 0 h(n = 3, P < 0.05); the ROS levels were 616.53% ± 79.78%, 974.81% ± 102.40%, and 1330.94% ± 93.09% vs 603.74% ± 71.99%(n = 3, P < 0.05); the MMP values were 2.07 ± 0.05, 1.78 ± 0.29, and 1.16 ± 0.25 vs 2.55 ± 0.42(n = 3, P < 0.05); caspase-3 activity was significantly higher compared to the control(5492.3 ± 321.1, 6562.0 ± 381.3, and 8695.7 ± 449.1 vs 2330.0 ± 121.9), but the caspase activity of the non-tumor cell line L-O2 was not different from that of the control. With the addition of the caspase-3 inhibitor(Ac-DEVD-CHO), caspase-3 activity was significantly decreased compared to the control group(1067.0 ± 132.5 U/g vs 8695.7 ± 449.1 U/g). The expression of the Cyt C, Endo G, and AIF proteins in SGC-7901 cells was significantly higher than those in the control(P < 0.05), while the expression of the Smac/Diablo protein was significantly lower than the control group after melittin exposure(P < 0.01). Ac-DEVD-CHO did not, however, have any effect on the expression of caspase-8 and FAS in the SGC-7901 cells.CONCLUSION: Melittin can induce apoptosis of human gastric cancer(GC) cells through the mitochondria pathways, and it may be a potent agent in the treatment of human GC.展开更多
This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and th...This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and the underlying mechanism.Two hundred SD rats were randomly divided into four groups:normal group,sham-operated group,model group and EA+TMS group,and each group was then divided into five sub-groups in terms of the different time to start treatment post operation:6,12,24,48 and 72 h.Cerebral infarction models were established in the model and the EA+TMS groups by left middle cerebral artery occlusion/reperfusion (MCAO/R).After treatment for 14 d,the Morris water maze test was applied to examine the spatial learning and memory abilities of rats.In infarcted area,the expression of caspase-3 was immunohistochemically detected,and real-time fluorescent quantitative PCR was used to measure the expression of Bcl-2 mRNA.The results showed that in EA+TMS group compared with model group at the same treatment time windows,the escape latency was substantially shortened,the expression of caspase-3 was considerably decreased and the expression level of Bcl-2 mRNA significantly increased (P<0.05).In the EA+TMS sub-groups,the escape latency was shortest,the expression level of caspase-3 lowest,and the expression level of Bcl-2 mRNA highest at the treatment time window of 24 h.It was concluded that EA combined with TMS can promote neurological function of rats with cerebral infarction by increasing the expression level of Bcl-2 mRNA and decreasing the expression of caspase-3.The best time window is 24 h after perfusion treatment to ischemia.展开更多
Aim: Diabetes mellitus is a metabolic disorder leading to hyperglycemia and exhibiting altered fat and protein metabolism. Diabetes altered cellular microenvironment caused myriad untoward effects. Periodontitis is ch...Aim: Diabetes mellitus is a metabolic disorder leading to hyperglycemia and exhibiting altered fat and protein metabolism. Diabetes altered cellular microenvironment caused myriad untoward effects. Periodontitis is chronic inflammatory disease. Diabetes and periodontitis had higher prevalence in populations. The objective studied the relationship between diabetes and periodontitis associated with cell apoptosis and the influence of diabetes enhanced inflammation on apoptosis and periodontitis. Methods: This paper studied and analyzed the papers which published in the worldwide associated with the influence of diabetes enhanced inflammation on cell apoptosis and periodontitis, and reviewed the probably mechanism associated with apoptosis. Results: Diabetes induced hyperglycemia enhanced inflammation related to cell apoptosis. Periodontitis had a higher morbidity on diabetes patients. Periodontal intervention may be benefit to controlling the diabetes. The bidirectional efficiency happened between diabetes and periodontitis. Anti-apoptotic and anti-inflammation option can improve the therapeutic effects on diabetes and periodontitis. The finding included following several aspects. 1) Advanced glycation end products enhanced inflammatory response;2) Hyperglycemia induced cell apoptosis;3) inflammatory cytokines caused cell apoptosis;4) Mutuality between cell apoptosis and periodontitis;5) Diabetes induce periodontitis and bone loss;6) Periodontitis induced insulin resistance. 7) TNFα induce prostaglandins elicited cell apoptosis;8) periodontal therapies had effects on diabetes. Conclusion: Diabetes can enhance inflamemation leading to apoptosis and periodontitis. Effective periodontal therapy and control glucose may produce better effects on diabetes or periodontitis. Further research required to investigate the bidirectional mechanism between diabetes and periodontitis.展开更多
基金supported by the National Key R&D Program of China(2022YFD1600903)the National Natural Science Foundation of China(32072693)the College Students’Innovative Entrepreneurial Training Plan Program(202110307028).
文摘Background Atresia and degeneration,a follicular developmental fate that reduces female fertility and is triggered by granulosa cell(GC)apoptosis,have been induced by dozens of miRNAs.Here,we report a miRNA,miR-423,that inhibits the initiation of follicular atresia(FA),and early apoptosis of GCs.Results We showed that miR-423 was down-regulated during sow FA,and its levels in follicles were negatively correlated with the GC density and the P4/E2 ratio in the follicular fluid in vivo.The in vitro gain-of-function experiments revealed that miR-423 suppresses cell apoptosis,especially early apoptosis in GCs.Mechanically speaking,the miR-423 targets and interacts with the 3’-UTR of the porcine SMAD7 gene,which encodes an apoptosis-inducing factor in GCs,and represses its expression and pro-apoptotic function.Interestingly,FA and the GC apoptosis-related lncRNA NORHA was demonstrated as a ceRNA of miR-423.Additionally,we showed that a single base deletion/insertion in the miR-423 promoter is significantly associated with the number of stillbirths(NSB)trait of sows.Conclusion These results demonstrate that miR-423 is a small molecule for inhibiting FA initiation and GC early apoptosis,suggesting that treating with miR-423 may be a novel approach for inhibiting FA initiation and improving female fertility.
基金supported by the Shandong Provincial Natural Science Foundation of China(ZR2019MH096).
文摘Background:Dihydroartemisinin(DHA)is reported to be a potential anticancer agent,and the mechanisms underlying the effects of DHA on diffuse large B cell lymphoma however are still obscure.This study aimed to assess the antitumor effect of DHA on diffuse large B cell lymphoma cells and to determine the potential underlying mechanisms of DHA-induced cell apoptosis.Methods:Here,the Cell Counting Kit 8 assay was conducted to study cell proliferation.We performed Annexin V-FITC/propidium iodide staining,real-time polymerase chain reaction,and western blot analysis to analyze cell apoptosis and potential molecular mechanisms.Results:The results showed that DHA substantially suppressed cell proliferation and induced cell apoptosis in vitro in a time-and concentration-dependent fashion.Moreover,STAT3 activity could be inhibited after stimulation with DHA.Conclusion:These results imply that the underlying anti-tumoral effect of DHA may increase apoptosis in diffuse large B cell lymphoma cells via the STAT3 signaling pathway.In addition,DHA might be an effective drug for diffuse large B cell lymphoma therapy.
基金Supported by the National Natural Science Foundation of China(No.81771499)the Natural Science Foundation of Hebei Province,China(No.H2018206099,No.H2021206460)。
文摘AIM:To determine whether an antisense RNA corresponding to the human Alu transposable element(Aluas RNA)can protect human lens epithelial cells(HLECs)from methylglyoxal-induced apoptosis.METHODS:Cell counting kit-8(CCK-8)and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assays were used to assess HLEC viability.HLEC viability/death was detected using a Calcein-AM/PI double staining kit;the annexin V-FITC method was used to detect HLEC apoptosis.The cytosolic reactive oxygen species(ROS)levels in HLECs were determined using a reactive species assay kit.The levels of malondialdehyde(MDA)and the antioxidant activities of total-superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px)were assessed in HLECs using their respective kits.RT-q PCR and Western blotting were used to measure m RNA and protein expression levels of the genes.RESULTS:Aluas RNA rescued methylglyoxal-induced apoptosis in HLECs and ameliorated both the methylglyoxalinduced decrease in Bcl-2 m RNA and the methylglyoxalinduced increase in Bax m RNA.In addition,Aluas RNA inhibited the methylglyoxal-induced increase in Alu sense RNA expression.Aluas RNA inhibited the production of ROS induced by methylglyoxal,restored T-SOD and GSHPx activity,and moderated the increase in MDA content after treatment with methylglyoxal.Aluas RNA significantly restored the methylglyoxal-induced down-regulation of Nrf2 gene and antioxidant defense genes,including glutathione peroxidase,heme oxygenase 1,γ-glutamylcysteine synthetase and quinone oxidoreductase 1.Aluas RNA ameliorated methylglyoxal-induced increases of the m RNA and protein expression of Keap1 that is the negative regulator of Nrf2.CONCLUSION:Aluas RNA reduces apoptosis induced by methylglyoxal by enhancing antioxidant defense.
文摘White tea encompasses a number of teas unique to eastern Fujian in China. Although white tea extracts have been reported to result in cancer cell apoptosis, to date, few studies have evaluated the mechanism of such apoptotic induction. A transcription factor that plays a critical role in cell apoptosis, NF-κB p65, is also likely critical by which white tea extracts induce cancer cell apoptosis. In this study, white tea aqueous extract (WTAE) was added to BEL-7402 and Hela cell media and NF-κB p65 activation was evaluated using western blotting and immunofluorescence. Results revealed that the phosphorylation of IKBα and p65 decreased in both cell lines after WTAE treatment. And the nuclear translocation of NF-κB p65 in both cell lines was also reduced with the WTAE treatment. NF-κB p65 inhibition was noted to accelerate apoptosis. Our findings suggest that NF-κB p65 was an important modulator in WTAE-induced apoptotic signal transduction and it acted as a negative regulator of apoptotic induction in BEL-7402 and Hela cancer cell lines.
基金supported by the National Natural Science Foundation of China,Nos.30772368(to DH),81371034(to XH)the Key Project of Natural Science Foundation of Shaanxi Province,No.2017JZ025(to DH).
文摘The retinal ganglion cells of the optic nerve have a limited capacity for self-repair after injury.Valproate is a histone deacetylase inhibitor and multitarget drug,which has been demonstrated to protect retinal neurons.In this study,we established rat models of optic nerve-crush injury and injected valproate into the vitreous cavity immediately after modeling.We evaluated changes in the ultrastructure morphology of the endoplasmic reticulum of retinal ganglion cells over time via transmission electron microscope.Immunohistochemistry and western blot assay revealed that valproate upregulated the expression of the endoplasmic reticulum stress marker glucose-regulated protein 78 and downregulated the expression of transcription factor C/EBP homologous protein,phosphorylated eukaryotic translation initiation factor 2α,and caspase-12 in the endoplasmic reticulum of retinal ganglion cells.These findings suggest that valproate reduces apoptosis of retinal ganglion cells in the rat after optic nerve-crush injury by attenuating phosphorylated eukaryotic translation initiation factor 2α-C/EBP homologous protein signaling and caspase-12 activation during endoplasmic reticulum stress.These findings represent a newly discovered mechanism that regulates how valproate protects neurons.
文摘BACKGROUND As an active ingredient derived from Dioscorea zingiberensis C.H.Wright,deltonin has been reported to show anti-cancer effects in a variety of malignancies.AIM To investigate the role and mechanism of action of deltonin in promoting gastric carcinoma(GC)cell apoptosis and chemosensitivity to cisplatin.METHODS The GC cell lines AGS,HGC-27,and MKN-45 were treated with deltonin and then subjected to flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-3,5-diphenyltet-razolium bromide assays for cell apoptosis and viability determination.Western blot analysis was conducted to examine alterations in the expression of apoptosis-related proteins(Bax,Bid,Bad,and Fas),DNA repair-associated proteins(Rad51 and MDM2),and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of the rapamycin(PI3K/AKT/mTOR)and p38-mitogen-activated protein kinase(MAPK)axis proteins.Additionally,the influence of deltonin on GC cell chemosensitivity to cisplatin was evaluated both in vitro and in vivo.RESULTS Deltonin treatment weakened viability,enhanced apoptosis,and dampened DNA repair in GC cell lines in a dose-dependent pattern.Furthermore,deltonin mitigated PI3K,AKT,mTOR,and p38-MAPK phosphorylation.HS-173,an inhibitor of PI3K,attenuated GC cell viability and abolished deltonin inhibition of GC cell viability and PI3K/AKT/mTOR and p38-MAPK pathway activation.Deltonin also promoted the chemosensitivity of GC cells to cisplatin via repressing GC cell proliferation and growth and accelerating apoptosis.CONCLUSION Deltonin can boost the chemosensitivity of GC cells to cisplatin via inactivating p38-MAPK and PI3K/AKT/mTOR signaling.
基金supported by the Central Government Supports Local College Reform and Development Fund Talent Training Projects[Grant Number 2020GSP16]the Heilongjiang Touyan Innovation Team Program[Grant Number 2019HTY078].
文摘Marine collagen peptides(MCPs)are natural products prepared by hydrolyzing marine collagen protein through a variety of chemical methods or enzymes.MCPs have a range of structures and biological activities and are widely present in marine species.MCPs also have a small molecular weight,are easily modified,and absorbed by the body.These properties have attracted great interest from researchers studying antioxidant,anti-tumor,and anti-aging activities.MCPs of specific molecular weights have significant anti-tumor activity and no toxic side effects.Thus,MCPs have the potential use as anti-cancer adjuvant drugs.Free radicals produced by oxidation are closely related to human aging,cancer,arteriosclerosis,and other diseases,but their relationship with cancer is not well known.In this review,we focus on the antioxidant properties of MCPs in the treatment of cancer,highlighting their antioxidant molecular structure and potential for clinical practice.
基金Supported by National Key R&D Program of China(No.2021YFC2702103,No.2021YFC2702100)the National Natural Science Foundation of China(No.82104937)the Key R&D Program of Shandong Province(No.2019GSF108252).
文摘AIM:To investigate the effect of electroacupuncture(EA)on the mitochondria-dependent apoptotic signaling pathway in the ciliary muscle of guinea pigs with negative lensinduced myopia(LIM).METHODS:Guinea pigs were randomly divided into normal control(NC)group,LIM group,LIM+SHAM acupoint(LIM+SHAM)group,and LIM+EA group.Animals in the NC group received no intervention,while those in other three groups were covered with-6.0 diopter(D)lenses on right eyes.Meanwhile,animals in the LIM+EA group received EA at Hegu(LI4)combined with Taiyang(EX-HN5)acupoints,while those in the LIM+SHAM group were treated at sham points.After treatments for 1,2,and 4wk,morphological changes in ciliary muscles were observed with hematoxylin and eosin(H&E)staining and nick end labeling(TUNEL),and the expression of the mitochondrial apoptotic signaling pathway-related molecules in ciliary muscles was measured by real-time quantitative polymerase chain reaction(qPCR)and Western blot.Additionally,the adenosine triphosphate(ATP)contents were also determined in ciliary muscles.RESULTS:Axial length increased significantly in the LIM and LIM+SHAM groups and decreased in the LIM+EA group.The ciliary muscle fibers were broken and destroyed in both LIM and LIM+SHAM groups,whereas those in the LIM+EA group improved significantly.TUNEL assay showed the number of apoptotic cells increased in the LIM and LIM+SHAM groups,whereas reduced in the LIM+EA group.ATP contents showed a significant decrease in the LIM and LIM+SHAM groups,whereas increased after EA treatment.Compared with the NC group,the dynamin-related protein 1(DRP1),Caspase3,and apoptotic protease activator 1(APAF1)levels were significantly increased in the LIM group and decreased in the LIM+EA group.CONCLUSION:The results provide evidence of EA inhibiting the development of myopia by regulating the mitochondrial apoptotic signaling pathway.
文摘BACKGROUND Gastric adenocarcinoma(GAC)mortality rates have remained relatively changed over the past 30 years,and it continues to be one of the leading causes of cancerrelated death.AIM To search for novel miRNAs related to GAC prognosis and further investigate the effect of miR-96-5p on MGC-803 cells.METHODS The miRNA expression profile data of GAC based on The Cancer Genome Atlas were obtained and used to screen differently expressed miRNAs(DEMs)and DEMs related to GAC prognosis.Then,the expression of DEMs related to GAC prognosis was identified in GAC tumor samples and adjacent normal samples by qRT-PCR.The target gene,ZDHHC5,of miR-96-5p was predicted using TargetScan,miRTarBase,and miRDB databases and confirmed by luciferase reporter assay.Furthermore,MGC-803 cells were transfected with inhibitor NC,miR-96-5p inhibitor,si-ZDHHC5,or miR-96-5p inhibitor+si-ZDHHC5,and then cell apoptosis was detected by flow cytometry.The expression of ZDHHC5,Bcl-2,and COX-2 was detected using western blotting.RESULTS A total of 299 DEMs and 35 DEMs related to GAC prognosis were screened based on The Cancer Genome Atlas.Then compared with adjacent normal samples,the levels of miR-96-5p,miR-222-5p,and miR-652-5p were remarkably increased,while miR-125-5p,miR-145-3p,and miR-379-3p levels were reduced in GAC tumor samples(P<0.01),which were consistent with bioinformatics analysis.Furthermore,ZDHHC5 was defined as a direct target gene of miR-96-5p.miR-96-5p inhibition increased the number of apoptotic cells as well as promoted the expression of ZDHHC5,Bcl-2,and COX-2 in MGC-803 cells(P<0.01).After ZDHHC5 inhibition,the number of apoptotic cells and the expression of ZDHHC5,Bcl-2,and COX-2 were reduced.The addition of an miR-96-5p inhibitor partly reversed these effects(P<0.01).CONCLUSION Our findings identified six miRNAs related to GAC prognosis and suggested that downregulated miR-96-5p might induce cell apoptosis via upregulating ZDHHC5 expression in MGC-803 cells.
基金Scientific and Technological Research Developing Program of Shaanxi Province, No. 2007K15-01
文摘BACKGROUND: Neuronal apoptosis in perihematomal brain tissues following intracerebral hemorrhage is strongly related to the formation of a compound signal pathway between nerve growth factor precursor (proNGF), p75NTR, and sortilin receptor. Sortilin acts as a co-receptor and molecular switch governing the p75NTR-mediated pro-apoptotic signal induced by proNGF. OBJECTIVE: To investigate proNGF and sortilin expressions in perihematomal brain tissues following intracerebral hemorrhage, and to study the effects of proNGF and sortilin on secondary cell apoptosis. DESIGN, TIME AND SETTING: A paired, comparison study was performed at the Laboratory of Histology and Embryology, Xi'an Jiaotong University from October 2007 to September 2008. MATERIALS: Brain tissue samples were obtained from 15 patients with intracerebral hemorrhage, who were treated at the Department of Neurosurgery, First Affiliated Hospital, Medical College of Xi'an Jiaotong University from October 2007 to March 2008. Rabbit anti-proNGF polyclonal antibody was provided by Chemicon, USA; rabbit antisortilin polyclonal antibody by Abcam, UK; and TUNEL kit by Promega, USA. METHODS: Perihematomal brain tissues selected 0.5 cm from the hemorrhage area were considered to be the hemorrhage group, while brain tissues from the middle temporal gyrus served as the control group. MAIN OUTCOME MEASURES: Histopathological changes were detected using hematoxylin-eosin staining, cell apoptosis was determined using the TUNEL method, and proNGF and sortilin ex- pressions were determined using immunohistochemistry. RESULTS: Edema was clearly observed in perihematomal brain tissues, and infiltration of inflammatory cells was visible, with the presence of irregular and necrotic bodies. The apoptotic rate in the hemorrhage group was significantly greater than in the control group (P < 0.01). Moreover, sortilin expression significantly increased (P < 0.01), but proNGF expression remained unchanged (P > 0.05). Correlation analysis suggested that sortilin expression positively correlated with apoptosis (rs = 0.648, P < 0.01). CONCLUSION: proNGF expression was stable, but sortilin expression increased in perihematomal brain tissues following intracerebral hemorrhage, suggesting that sortilin acted as a co-receptor and molecular switch to govern p75NTR-mediated cell apoptosis.
基金This work was supported by the National Natural Science Foundation of China(32072693 and 31630072)the Qing Lan Project of Jiangsu Province(2020).
文摘Background:Follicular atresia has been shown to be strongly associated with a low follicle utilization rate and female infertility,which are regulated by many factors such as microRNAs(miRNAs),which constitute a class of noncoding RNAs(ncRNAs).However,little is known about long noncoding RNAs(lncRNAs),which constitute another ncRNA family that regulate follicular atresia.Results:A total of 77 differentially expressed lncRNAs,including 67 upregulated and 10 downregulated lncRNAs,were identified in early atretic follicles compared to healthy follicles by RNA-Sequencing.We characterized a noncoding RNA that was highly expressed in atretic follicles(NORHA).As an intergenic lncRNA,NORHA was one of the upregulated lncRNAs identified in the atretic follicles.To determine NORHA function,RT-PCR,flow cytometry and western blotting were performed,and the results showed that NORHA was involved in follicular atresia by influencing GC apoptosis with or without oxidative stress.To determine the mechanism of action,bioinformatics analysis,luciferase reporter assay and RNA immunoprecipitation assay were performed,and the results showed that NORHA acted as a‘sponge’,that directly bound to the miR-183-96-182 cluster,and thus prevented its targeted inhibition of FoxO1,a major sensor and effector of oxidative stress.Conclusions:We provide a comprehensive perspective of lncRNA regulation of follicular atresia,and demonstrate that NORHA,a novel lncRNA related to follicular atresia,induces GC apoptosis by influencing the activities of the miR-183-96-182 cluster and FoxO1 axis.
基金supported by grants from the National Natural Science Foundation of China[NSFC31500618,awarded to GAO JNSFC82000253,awarded to WANG HY]sponsored by Shanghai Sailing Program[20YF1414000 to WANG HY]。
文摘Objective Epidemiological studies reveal that exposure to fine particulate matter(aerodynamic diameter≤2.5μm,PM_(2.5))increases the morbidity and mortality of respiratory diseases.Emerging evidence suggests that human circulating extracellular vesicles(EVs)may offer protective effects against injury caused by particulate matter.Currently,however,whether EVs attenuate PM_(2.5)-induced A549 cell apoptosis is unknown.Methods EVs were isolated from the serum of healthy subjects,quantified via nanoparticle tracking analysis,and qualified by the marker protein CD63.PM_(2.5)-exposed(50μg/mL)A549 cells were pretreated with 10μg/mL EVs for 24 h.Cell viability,cell apoptosis,and AKT activation were assessed via Cell Counting Kit-8,flow cytometry,and Western blot,respectively.A rescue experiment was also performed using MK2206,an AKT inhibitor.Results PM_(2.5)exposure caused a 100%in crease in cell apoptosis.EVs treatme nt reduced cell apoptosis by 10%,promoted cell survival,and inhibited the PM_(2.5)-induced upregulation of Bax/Bcl2 and cleaved caspase 3/caspase 3 in PM_(2.5)-exposed A549 cells.Moreover,EVs treatment reversed PM_(2.5)-induced reductions in p-AKT^(Thr308)and p-AKT^(Ser473).A KT inhibition attenuated the anti-apoptotic effect of EVs treatment on PM_(2.5)-exposed A549 cells.Conclusions EVs treatment promotes cell survival and attenuates PM_(2.5)-induced cell apoptosis via AKT phosphorylation.Human serum-derived EVs may be an efficacious novel therapeutic strategy in PM_(2.5)-induced lung injury.
基金supported by a grant from the Foundation of Most Advanced Group of Medical Scientists and Technicians of Shandong Province (2007GG30002014)
文摘BACKGROUND:Many Chinese herbs,especially herbal injections,have been shown to have anti-tumor effects in recent years.However,since most reports focus on the clinical effectiveness of these herbs,their mechanisms of action are not well understood.In this study,we assessed apoptosis in the hepatocellular carcinoma (HCC) cell line HepG2 induced by an injectable extract from the seed of Coix lacryma-jobi (Semen coicis,SC),and monitored the expression of Bcl-2 and caspase-8.METHODS:Injectable SC was applied to HepG2 cells at different concentrations and the cells were collected 12,24 and 48 hours later.5-fluorouracil was used as a positive control group,and fluorescence-activated cell-sorting cytometry was used to measure the apoptosis rate of HepG2 cells and the expression of Bcl-2 and caspase-8 proteins.RESULTS:SC induced apoptosis in HepG2 cells in a concentration and time-dependent manner,and the expression of caspase-8 was elevated and prolonged.However,it did not significantly influence the expression of Bcl-2.CONCLUSION:Injectable SC may induce apoptosis in HCC cells by regulating the expression of caspase-8.
基金supported by Wenzhou Science and Technology Project(Y20080087)
文摘Objective:To study the protective effect of resveratrol on lens epithelial cell apoptosis in diabetic cataract rat.Methods:A total of 84 Wistar rats were divided into 4 groups:12 in Group A(control group).24 in Group B(diabetic cataract group),24 in Group C(therapeutic-dose of resveratrol group) and 24 in Group D(low-dose of resveratrol group).Rats in Group B-D were given with60 mg/kg streptozotocin through intraperitoneal injection.Rats in Group C were given with 100mg/kg resveratrol and rats in Group D were given with 20 mg/kg resveratrol.The caspase-3expression levels and apoptosis ratios of LEC among each group were observed:the degrees of lens opacity in Group B-D after 12 weeks were compared.Results:There were significant differences in caspase-3 expression levels,apoptosis ratios of T.F.C among groups at 4 w,8 w and12 w(P<0.05).After 12 weeks,in Group B the degree of lens opacity was as follow:0(0.00%) in grade Ⅰ,3(37.50%) in grade Ⅱ,2(25.00%)in grade Ⅲ,2(25.00%)grade Ⅳ,and 1(12.50%) in gradeⅤ:in Group C:2(25.00%)in grade Ⅰ,4(50.00%) in grade Ⅱ.2(25.00%)in grade Ⅲ,0(0.00%)gradeⅣ,and 0(0.00%) in grade Ⅴ;in Group D:1(12.50%)in grade Ⅰ,4(50.00%) in grade Ⅱ,2(25.00%)in grade Ⅲ,1(12.50%) grade Ⅳ,and 0(0.00%) in grade Ⅴ.The.difference among Group B-D was statistically significant(P<0.05).Conclusions:Resveratrol has protective effect on lens epithelial cell apoptosis in diabetic cataract rat,and the effect is relative to its dose.
基金Supported by Grants from the American Heart Association, to Habib SL
文摘Apoptosis contributes to the development of diabetic nephropathy, but the mechanism by which high glucose induces apoptosis is not fully understood. Apoptosis of tubular epithelial cells is a major feature of diabetic kidney disease, and hyperglycemia triggers the generation of free radicals and oxidant stress in tubular cells. Hyperglycemia and high glucose in vitro also lead to apoptosis, a form of programmed cell death. High glucose similar to those seen with hyperglycemia in people with diabetes mellitus, lead to accelerated apoptosis, a form of programmed cell death characterized by cell shrinkage, chromatin condensation and DNA fragmentation, in variety of cell types, including renal proximal tubular epithelial cells.
文摘AIM:To investigate whether Melatonin has synergistic effects with Doxorubicin in the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and Bel-7402.METHODS:The synergism of Melatonin and Doxorubicin inhibited the cell growth and induced cell apoptosis in human hepatoma cell lines HepG2 and Bel-7402.Cell viability was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT)assay.Cell apoptosis was evaluated using TUNEL method and flow cytometry.Apoptosis-related protein Bax,Bcl-2 and caspase-3 expressions were measured by immunohistochemical staining.RESULTS:Treatment with Melatonin(10 -8 -10 -5 mol/L) alone had a dose-related inhibitory effect on cell proliferation but no cytotoxic effect on hepatoma cell lines HepG2 and Bel-7402.Interestingly,when combined with Doxorubicin,Melatonin significantly increased the effects of cell growth inhibition and cell apoptosis.Furthermore,TUNEL staining and flow cytometry revealed that cooperative apoptosis induction was associated with decreased expression of Bcl-2 as well as increased expression of Bax and Caspase3.CONCLUSION:The synergism of Melatonin and Doxorubicin inhibits hepatoma cell growth and induces cell apoptosis.
基金The design of the study and collection,analysis,and interpretation of data and in writing the manuscript were supported by the Specialized Research Fund for the Doctoral Program of Higher Education(20130008130001)
文摘Background: Heat stress is known to alter follicular dynamics and granulosa cell function and may contribute to the diminished reproductive efficiency commonly observed in mammals during the summer. Although several investigators have studied heat-induced ovarian injury, few reports have focused on the effects of chronic heat stress on ovarian function and the molecular mechanisms through which it induces ovarian injury.Methods: In Exp. 1, 48 female mice were assigned to a control or heat-stressed treatment. After exposure to a constant temperature of 25 °C for 7, 14, 21 or 28 d(n = 6) or to 42 °C for 3 h per d for 7, 14, 21 or 28 d(n = 6), the mice were euthanized and their ovaries were analyzed for follicular atresia, granulosa cell apoptosis, changes in the abundance of HSP70 protein and serum concentrations of estradiol. In Exp. 2, the expression of HSP70 and aromatase was quantified in antral follicles cultured in vitro at 37 or 42 °C for 24 h. In Exp. 3, granulosa cells from ovaries maintained at 37 or 41 °C for 2 h were analyzed for their expression of HSP70, Bim, caspase-3 and cleaved caspase-3.Results: In Exp. 1, body weight and food intake of heat-stressed mice decreased(P < 0.05) compared with control mice while the concentration of estradiol in serum was lower(P < 0.05) in heat-stressed mice than in control mice. Compared with control mice, the percentage of atretic follicles and the number of antral follicles with severe apoptotic signals were increased(P < 0.05) after 21 d of heat-stressed treatment. HSP70 protein was more abundant(P < 0.05) in heat-stressed mice than control mice. In Exp. 2, heat stress increased HSP70 and decreased aromatase proteins(P < 0.05) in antral follicles. In Exp. 3, TUNEL-positive granulosa cells from heat-stressed ovaries were observed concomitant with a significant increase in HSP70, Bim and cleaved caspase-3 protein.Conclusion: Heat-stress in mice decrease estradiol in serum and aromatase in antral follicles but increased number of atretic follicles and granulosa cell undergoing apoptosis which may explain the decreased fertility commonly observed in heat-stressed animals.
基金Supported by the Natural Science Foundation of ChinaNo.30801497+2 种基金No.81272537 and No.81472815the Natural Science Fund for Colleges and Universities in Jiangsu ProvinceNo.11KJD360003
文摘AIM: To investigate the apoptotic effects of melittin on SGC-7901 cells via activation of the mitochondrial signaling pathway in vitro.METHODS: SGC-7901 cells were stimulated by melittin, and its effect on proliferation and apoptosis of was investigated by methyl thiazolyl tetrazolium assay, morphologic structure with transmission electron microscopy, annexin-V/propidium iodide double-staining assay, measuring mitochondrial membrane potential(MMP) levels, and analyzing reactive oxygen species(ROS) concentrations were analyzed by flow cytometry. Cytochrome C(Cyt C), apoptosis-inducing factor(AIF), endonuclease G(Endo G), second mitochondria-derived activator of caspases(Smac)/direct IAP binding protein with low isoelectric point(Diablo), and FAS were analyzed by western blot. The expression of caspase-3 and caspase-8 was measured using activity assay kits.RESULTS: Melittin was incubated at 1.0, 2.0, 4.0, or 6.0 μg/m L for 1, 2, 4, 6, or 8 h and showed a timeand concentration-dependent inhibition of SGC-7901 cell growth. Melittin induced SGC-7901 cell apoptosis, which was confirmed by typical morphological changes. Treatment with 4 μg/m L melittin induced early apoptosis of SGC-7901 cells, and the early apoptosis rates were 39.97% ± 3.19%, 59.27% ± 3.94%, and 71.50% ± 2.87% vs 32.63% ± 2.75% for 1, 2, and 4 h vs 0 h(n = 3, P < 0.05); the ROS levels were 616.53% ± 79.78%, 974.81% ± 102.40%, and 1330.94% ± 93.09% vs 603.74% ± 71.99%(n = 3, P < 0.05); the MMP values were 2.07 ± 0.05, 1.78 ± 0.29, and 1.16 ± 0.25 vs 2.55 ± 0.42(n = 3, P < 0.05); caspase-3 activity was significantly higher compared to the control(5492.3 ± 321.1, 6562.0 ± 381.3, and 8695.7 ± 449.1 vs 2330.0 ± 121.9), but the caspase activity of the non-tumor cell line L-O2 was not different from that of the control. With the addition of the caspase-3 inhibitor(Ac-DEVD-CHO), caspase-3 activity was significantly decreased compared to the control group(1067.0 ± 132.5 U/g vs 8695.7 ± 449.1 U/g). The expression of the Cyt C, Endo G, and AIF proteins in SGC-7901 cells was significantly higher than those in the control(P < 0.05), while the expression of the Smac/Diablo protein was significantly lower than the control group after melittin exposure(P < 0.01). Ac-DEVD-CHO did not, however, have any effect on the expression of caspase-8 and FAS in the SGC-7901 cells.CONCLUSION: Melittin can induce apoptosis of human gastric cancer(GC) cells through the mitochondria pathways, and it may be a potent agent in the treatment of human GC.
基金supported by a grant from the National Natural Science Foundation of China (No.30640010)
文摘This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and the underlying mechanism.Two hundred SD rats were randomly divided into four groups:normal group,sham-operated group,model group and EA+TMS group,and each group was then divided into five sub-groups in terms of the different time to start treatment post operation:6,12,24,48 and 72 h.Cerebral infarction models were established in the model and the EA+TMS groups by left middle cerebral artery occlusion/reperfusion (MCAO/R).After treatment for 14 d,the Morris water maze test was applied to examine the spatial learning and memory abilities of rats.In infarcted area,the expression of caspase-3 was immunohistochemically detected,and real-time fluorescent quantitative PCR was used to measure the expression of Bcl-2 mRNA.The results showed that in EA+TMS group compared with model group at the same treatment time windows,the escape latency was substantially shortened,the expression of caspase-3 was considerably decreased and the expression level of Bcl-2 mRNA significantly increased (P<0.05).In the EA+TMS sub-groups,the escape latency was shortest,the expression level of caspase-3 lowest,and the expression level of Bcl-2 mRNA highest at the treatment time window of 24 h.It was concluded that EA combined with TMS can promote neurological function of rats with cerebral infarction by increasing the expression level of Bcl-2 mRNA and decreasing the expression of caspase-3.The best time window is 24 h after perfusion treatment to ischemia.
文摘Aim: Diabetes mellitus is a metabolic disorder leading to hyperglycemia and exhibiting altered fat and protein metabolism. Diabetes altered cellular microenvironment caused myriad untoward effects. Periodontitis is chronic inflammatory disease. Diabetes and periodontitis had higher prevalence in populations. The objective studied the relationship between diabetes and periodontitis associated with cell apoptosis and the influence of diabetes enhanced inflammation on apoptosis and periodontitis. Methods: This paper studied and analyzed the papers which published in the worldwide associated with the influence of diabetes enhanced inflammation on cell apoptosis and periodontitis, and reviewed the probably mechanism associated with apoptosis. Results: Diabetes induced hyperglycemia enhanced inflammation related to cell apoptosis. Periodontitis had a higher morbidity on diabetes patients. Periodontal intervention may be benefit to controlling the diabetes. The bidirectional efficiency happened between diabetes and periodontitis. Anti-apoptotic and anti-inflammation option can improve the therapeutic effects on diabetes and periodontitis. The finding included following several aspects. 1) Advanced glycation end products enhanced inflammatory response;2) Hyperglycemia induced cell apoptosis;3) inflammatory cytokines caused cell apoptosis;4) Mutuality between cell apoptosis and periodontitis;5) Diabetes induce periodontitis and bone loss;6) Periodontitis induced insulin resistance. 7) TNFα induce prostaglandins elicited cell apoptosis;8) periodontal therapies had effects on diabetes. Conclusion: Diabetes can enhance inflamemation leading to apoptosis and periodontitis. Effective periodontal therapy and control glucose may produce better effects on diabetes or periodontitis. Further research required to investigate the bidirectional mechanism between diabetes and periodontitis.
