OBJECTIVE:To examine the effects of moxibustion on myocardial injury and myocardial metabolomics in rats with rheumatoid arthritis(RA)based on the transforming growth factor beta1(TGF-β1)/Smads signaling pathway.METH...OBJECTIVE:To examine the effects of moxibustion on myocardial injury and myocardial metabolomics in rats with rheumatoid arthritis(RA)based on the transforming growth factor beta1(TGF-β1)/Smads signaling pathway.METHODS:One hundred rats were treated with saline[normal control(NC)group]or complete Freund’s adjuvant(CFA)by right plantar injection for the RA model group,and the latter were randomly divided into 4 groups.Tripterygium wilfordii polyglycoside tablets(雷公藤多苷片,TPT)have anti-inflammatory and are widely used in the clinical treatment of RA,therefore serving as a positive control group.Three days post injection rats were given TPT tablet(TPT group),acupuncture therapy(APT group),and moxibustion treatment(MOX group)for 15 consecutive days,while NC group and model group were equally grasped and fixed and received normal saline.Rat joint swelling scores and arthritis index(AI)were evaluated in each group before the CFA challenge,therapy and after receiving therapy.Myocardial ultrastructure was observed by electron microscope.Enzyme-linked immunosorbent assay was used to detect cardiac troponin I(cTnI)levels in rat myocardial tissue.Quantitative reverse transcription polymerase chain reaction and Western blotting analysis were used to measure the mRNA and protein levels of TGF-βsignaling molecules including TGF-β1,Smad2,Smad3,Smad4,and Smad7.Myocardial metabolomics was analyzed using gas chromatography-mass spectrometer.RESULTS:Compared with model group,RA model rats receiving TPT,acupuncture,or moxibustion therapy all showed reduced joint swelling scores and AI(all P<0.01)and improved myocardial damage,whereas rats treated with moxibustion were found to be more marked.Consistently,the expressions of cTnI,TGF-β1,Smad2,Smad3,and Smad4 were found to be elevated in model rat group in contrast to NC rats and were significantly downregulated in TPT,APT and MOX group when compared with model group,while the levels of Smad7 showed the opposite result(all P<0.01).Moreover,the dissection of metabolomics suggested a novel metabolite biomarker panel including D-Xylulose 5-phosphate,dihydroxyacetone phosphate,arachidonic acid,etc was defined and implicated in amino acid,glucose,and fatty acid metabolic processes as revealed by principal component analysis and partial least squares discriminant analysis.CONCLUSION:Moxibustion prevents RA-induced inflammatory response and offers potent therapeutic effects on myocardial dysfunctions.The protective effects might be associated with its role in TGF-β1 inactivation and metabolic reprogramming.展开更多
OBJECTIVE:The aim of this study was to investigate the protective effects of Tuina(a traditional Chinese massage therapy)on intervertebral disc(IVD)degeneration and the regulatory mechanisms of the transforming growth...OBJECTIVE:The aim of this study was to investigate the protective effects of Tuina(a traditional Chinese massage therapy)on intervertebral disc(IVD)degeneration and the regulatory mechanisms of the transforming growth factor-β1(TGF-β1)/small mothers against decapentaplegic(Smad)signaling pathway.METHODS:Thirty New Zealand white rabbits were randomized into five groups:the control group,model group,model+Tuina group(Tuina group),model+TGF-β1 group(TGF-β1 group),and model+TGF-β1 inhibitor SB431542 group(SB431542 group).The model was established by posterolateral annulus fibrosus puncturing(AFP).Recombinant TGF-β1 and inhibitor SB431542 was injected into the TGF-β1 group and SB431542 group with a microsyringe,respectively.The rabbits in the Tuina group received Tuina treatment along the bladder meridian for 4 weeks.Magnetic resonance imaging(MRI)was performed on rabbits before AFP and after 4 weeks of intervention.Lumbar IVDs(L2-L3 to L4-L5)were harvested after intervention.Histopathological changes in the IVDs were measured by hematoxylin and eosin(HE)staining.Type I collagen was analyzed by immunohistochemistry detection.The expression level of matrix metalloproteinase-3(MMP3)was determined by enzyme-linked immunosorbent assay.Cell apoptosis was evaluated by terminal deoxynucleotidyl transferasemediated nick end labeling and Western blotting.Realtime polymerase chain reaction and Western blotting were used to analyze the expression of TGF-β1 and Smad2/3/4 and a disintegrin and metalloproteinase with thrombospondin motifs 5.RESULTS:Posterolateral AFP induced IVD degeneration in rabbits with histopathological damage and noticeable changes in MRI images.Tuina alleviated histopathological changes and reversed the expression of extracellular matrix degeneration-related molecules and apoptosis-related proteins.Furthermore,AFP induced the activation of TGF-β1 and Smad2/3/4,whereas Tuina therapy markedly reduced the protein expression of Smad2/3 and the gene expression of TGF-β1 and Smad2/3/4.Additionally,the TGF-β1/Smad signaling pathway was activated in the TGF-β1 group,while the TGF-β1/Smad signaling pathway was inhibited in the SB431542 group.CONCLUSION:Posterolateral AFP induced disc degeneration as determined by MRI assessment and histological analysis.Tuina alleviated disc degeneration,possibly by inhibiting the fibrotic response mediated by the TGF-β1/Smad pathway,thus alleviating extracellular matrix degeneration and reducing cell apoptosis.展开更多
OBJECTIVE:To explore the effect of electroacupuncture(EA)on rats with chronic fallopian tube inflammation and its potential mechanisms.METHODS:Thirty-six female Sprague-Dawley rats were divided into Control,Model and ...OBJECTIVE:To explore the effect of electroacupuncture(EA)on rats with chronic fallopian tube inflammation and its potential mechanisms.METHODS:Thirty-six female Sprague-Dawley rats were divided into Control,Model and EA groups.The pathological morphology of the fallopian tubes was observed by hematoxylin-eosin(HE)and Masson staining.The results of transforming growth factor-β1(TGF-β1),P38 mitogen-activated protein kinase(MAPK),phosphorylation(p)-p38MAPK in rat oviduct tissues were detected by immunohistochemistry.Results of P38MAPK,p-P38MAPK and TGF-β1 in rat oviduct tissues were detected by immunofluorescence.The expression level of p38MAPK,p-P38MAPK,TGF-β1 protein in rats was detected by Western blot.Quantitative real-time polymerase chain reaction(RT-q PCR)was used to detect m RNA expression levels of TGF-β1.RESULTS:It found that collagen fibers counts decreased significantly in EA group compared to Model group.The phosphorylation of P38MAPK in EA group was significantly reduced compared to Model group.The serum TGF-β1 expressions in EA group increased decreased significantly.CONCLUSION:Electroacupuncture was able to attenuate chronic salpingitis through down-regulating TGF-β1/MAPK signaling pathway.展开更多
OBJECTIVE:To investigate the efficacy of Shenweifang(SWF)-containing serum on transforming growth factor(TGF)-β1–induced fibroblast-myofibroblast transition in normal rat kidney interstitial fibroblast cells(NRK-49 ...OBJECTIVE:To investigate the efficacy of Shenweifang(SWF)-containing serum on transforming growth factor(TGF)-β1–induced fibroblast-myofibroblast transition in normal rat kidney interstitial fibroblast cells(NRK-49 F).METHODS:Sprague-Dawley rats were gavaged with one of five solutions:(a)saline;(b)saline plus low-dose SWF;(c)saline plus medium-dose SWF;(d)saline plus highdose SWF;and(e)saline plus valsartan.NRK-49 F cells were treated with TGF-β1 and cultured using serum from the gavaged rats.RESULTS:TGF-β1 treatment increased the expression ofα-smooth muscle actin,proliferating cell nuclear antigen,collagenⅠ,Smad3,mitogen-activated protein kinase(MAPK)10,and c-Jun N-terminal kinase(JNK)3 and induced abnormalities in cell morphology,cell cycle progression,and cell proliferation.CONCLUSIONS:SWF-or valsartan-containing serum corrected(or partially corrected)TGF-β1–induced abnormal changes in this in vitro system.SWF-containing serum reversed abnormalities in morphology,cell cycle progression,and proliferation in TGF-β1–treated NRK-49F cells,probably by blocking the TGF-β1/Smads and TGF-β1/MAPK/JNK pathways.展开更多
OBJECTIVE: To investigate the effect of Huangqi decoction( 黄芪汤) on renal interstitial fibrosis and its association with the transforming growth factor-β1(TGF-β1)/mitogen-activated protein kinase(MAPK) signaling p...OBJECTIVE: To investigate the effect of Huangqi decoction( 黄芪汤) on renal interstitial fibrosis and its association with the transforming growth factor-β1(TGF-β1)/mitogen-activated protein kinase(MAPK) signaling pathway. METHODS: 120 C57/BL mice were randomly divided into six groups: sham group, Enalapril(20 mg/kg) group, 5/6 nephrectomy model group, and 5/6 nephrectomy model plus Huangqicoction(0.12, 0.36 and 1.08 g/kg respectively) groups. Detecting 24hours urinary protein, blood pressure, serum creatinine, urea nitrogen content changes. Periodic Acid-Schiff stain(PAS) and Masson’s trichrome staining was used to observe the renal tissue pathological changes. Protein expression of TGF-β1, Phosphorylated P38 mitogen activated protein kinases(P-P38), Phosphorylated c-jun N-terminal kinase(P-JNK), Phosphorylated extracellular regulated proteinhnase(PERK), Fibroblast-specific protein-1(FSP-1), Alpha smooth muscle actin(α-SMA), Type Ⅲ collagen(Collagen Ⅲ), Connective tissue growth factor(CTGF),Bcl-2 Assaciated X protein(Bax) and B cell lymphoma 2(Bcl-2) were measured with western blot and immunohistochemical. RESULTS: Both Huangqi decoction and Enalapril improved the kidney function, 24 h urinary protein and the fibrosis in 5/6 nephrectomy mice, Huangqi decoction downregulated the expressions of TGF-β1, FSP-1, α-SMA, Collagen Ⅲ and CTGF in a dose-dependent manner, and it has a significant difference(P < 0.01) compared with model group.Huangqi decoction downregulated the expressions of P-P38, P-JNK, P-ERK and Bcl-2 in a dose-dependent manner, while upregulated the expression of Bax. CONCLUSIONS: The protective effect of Huangqi decoction for renal interstitial fibrosis in 5/6 nephrectomized mice via the inhibition of EpithelialMesenchymal Transitions and downregulating the TGF-β1/MAPK signaling pathway.展开更多
OBJECTIVE: To investigate the effect of Danggui Buxue Tang(DBT), a decoction from Traditional Chinese Medicine, on bleomycin-induced pulmonary fibrosis in rats, and to propose the possible underlying mechanism.METHODS...OBJECTIVE: To investigate the effect of Danggui Buxue Tang(DBT), a decoction from Traditional Chinese Medicine, on bleomycin-induced pulmonary fibrosis in rats, and to propose the possible underlying mechanism.METHODS: Forty male Sprague-Dawley rats were randomly divided into sham group, model group,prednisone group and DBT group. Pulmonary fibrosis rat model was established by intratracheal injection with bleomycin. Body weight and lung index were monitored. Histopathologic examination and collagen deposition were determined using Hematoxylin and eosin(HE) and Masson's trichrome staining. Immunohistochemistry staining was applied to observe the expression of alpha-smooth muscle actin(α-SMA). m RNA expression of α-SMA,collagen Ⅰ and collagen Ⅲ were measured by realtime fluorescence quantitative PCR(RT-q PCR). Inflammatory cytokines, including tumor necrosis factor alpha(TNF-α), interleukin-6(IL-6) and IL-1β in serum were detected by Enzyme-linked immunosorbent assay. Alkali hydrolysis method was conducted to investigate the content of hydroxyproline(HYP). Transforming growth factor-β1(TGF-β1),Smad3 and plasminogen activator inhibitor-1(PAI-1) protein level were examined by Western blot assay.RESULTS: DBT significantly reduced the severity of bleomycin-induced pulmonary fibrosis and inflammation as indicated by minimizing the lost of weight, and by lowering the levels of lung index, inflammation score, Ashcroft score, collagen volume fraction(%), HYP, α-SMA, collagen Ⅰ, collagen Ⅲ,TNF-α, IL-6, IL-1β, TGF-β1, Smad3 and PAI-1, consistent with the effect of prednisone.CONCLUSION: Our findings suggest that DBT is able to ameliorate the pulmonary fibrosis, the possible mechanism may involve inhibition of pulmonary inflammation and collagen deposition, possibly via suppressing TGF-β1/Smad3/PAI-1 signaling pathway.展开更多
OBJECTIVE:To explore the effects of Qingguang'an(青光安)containing serum on the expression levels of autophagy related genes in the transforming growth factor beta 1(TGF-β1)-activated human Tenon's fibroblast...OBJECTIVE:To explore the effects of Qingguang'an(青光安)containing serum on the expression levels of autophagy related genes in the transforming growth factor beta 1(TGF-β1)-activated human Tenon's fibroblasts(HTFs).METHODS:(a)Primary HTFs were stimulated by TGF-β1 and underwent immunohistochemistry,which established a cell model after Glaucoma filtration surgery(GFS).(b)The cell models were divided into 4 group:normal group(normal cells),model group(+TGF-β1),treatment group(+TGF-β1+medicated serum),and positive control group(TGF-β1+rapamycin).Then,Qingguang'an medicated serum with optimum concentration was added to the corresponding group.The autophagy positive cells were identified by the Cyto-ID autophagy detection kits under fluorescent microscope and Cytation 5 multifunctional instrument for cell imaging.And the mean fluorescence intensity of autophagy positive cells was determined by flow cytometry.The expression levels of autophagy related genes—Beclin-1,autophagy related gene 5(ATG-5),and microtubule-associated protein 1 light chain 3(LC-3Ⅱ)were detected by quantitative reverse transcription-polymerase chain reaction and Western blot analysis.RESULTS:Compared with the normal group and the model group,the relative mRNA expression levels of autophagy-related genes(Beclin-1,ATG-5 and LC-3Ⅱ)in the experimental group were notably increased(P<0.05,P<0.01),and with the extension of treatment time,it had an increasing trend(48 h was more obvious),which showed a certain time dependency;the protein expression levels of autophagy-related genes(Beclin-1,ATG-5,and LC-3Ⅱ)were significantly increased in the experimental group(P<0.05,P<0.01).With the prolongation of treatment time,there was an increasing trend(48 h was relatively obvious),and it revealed a certain time dependency CONCLUSION:The Qingguang'an medicated serum could up-regulate autophagy related genes(Beclin1,ATG5,and LC3Ⅱ)in the TGF-β1-activated HTFs.展开更多
Objective:To evaluate the effect of resveratrol against CCl4-induced nephrotoxicity.Methods:Forty-two male Wistar rats were divided into seven groups randomly.After six weeks,kidney weight,body weight,blood urea,serum...Objective:To evaluate the effect of resveratrol against CCl4-induced nephrotoxicity.Methods:Forty-two male Wistar rats were divided into seven groups randomly.After six weeks,kidney weight,body weight,blood urea,serum creatinine,oxidative stress markers,and gene expression of renal transforming growth factor-beta1(TGF-β1),TGF-βreceptor type 1(TGF-βR1)and Smad3 were determined.In addition,the protein level of TGF-β1 in the tissue lysate was measured.Results:Resveratrol had a protective role in renal tissue by the improvement of antioxidant balance and reduction of renal parameters such as creatinine and urea(P<0.001).In addition,the renal m RNA level of TGF-β1,TGF-βR1,Smad3,as well as the protein level of TGF-β1 were decreased in rats treated with resveratrol(P<0.001),compared to the CCl4 group.Conclusions:Overall,resveratrol shows a protective effect against nephrotoxicity in CCl4 treated rats by reducing oxidative stress status and modulating the TGF-βsignaling.展开更多
Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,an...Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,and its dynamic changes are closely related to the proliferation of endothelial cells,the recruitment of pericytes to blood vessels,and functional differentiation during embryonic vascular development.The pathophysiology of many cerebrovascular diseases is today understood as a disorder of endothelial cell function and an imbalance in the proportion of vascular cells.Indeed,mutations in ALK1 and its co-receptor endoglin are major genetic risk factors for vascular arteriovenous malformation.Many studies have shown that ALK1 is closely related to the development of cerebral aneurysms,arteriovenous malformations,and cerebral atherosclerosis.In this review,we describe the various roles of ALK1 in the regulation of angiogenesis and in the maintenance of cerebral vascular homeostasis,and we discuss its relationship to functional dysregulation in cerebrovascular diseases.This review should provide new perspectives for basic research on cerebrovascular diseases and offer more effective targets and strategies for clinical diagnosis,treatment,and prevention.展开更多
Stress, inflammation and Plasminogen activator inhibitor 1 (PAI-1) are key mechanisms throughout the development of aging, constituting a crossroad in the most frequent pathologies that accompany it. Among metabolic p...Stress, inflammation and Plasminogen activator inhibitor 1 (PAI-1) are key mechanisms throughout the development of aging, constituting a crossroad in the most frequent pathologies that accompany it. Among metabolic processes, obesity, metabolic syndrome and type 2 diabetes mellitus are included and Alzheimer’s disease among the neurodegenerative processes. Stress is a mechanism of defense of the organism against exogenous and endogenous actions called stressors. In the case of low intensity stimuli, the organism responds with actions aimed at a physiological adaptation (Homeostasis). On the other hand, when a high intensity (experimental level) or chronic stimulus (oxidative stress) is repeated, structural and functional changes are observed in different organs with activation of the hypothalamus-pituitary-adrenal axis, the renin angiotensin system and the sympathetic nervous system, stimulating the production of hormones that release cytokines with proin-flammatory/antiinflammatory properties that play an important role in the previously mentioned pathologies, as well as a marked increase in PAI-1, a gene regulated by stress and by cytokines, with manifest action at the origin of thromboembolic disease, so frequent in aging. The objective of this review is to highlight the importance of the binomial stress and PAI-1 in aging and in the pathologies that accompany it. Because PAI-1 is part of the pathology and complications in aging, some authors suggest the study of PAI-1 inhibitors to achieve its physiological levels, as part of the treatment of these diseases.展开更多
OBJECTIVE:To observe the efficacy of Danggui Buxue decoction(当归补血汤,DBD) on diabetic nephropathyinduced renal fibrosis in rats,and to study the possible mechanism.METHODS:Sixty male Goto Kakizaki(GK) rats were ran...OBJECTIVE:To observe the efficacy of Danggui Buxue decoction(当归补血汤,DBD) on diabetic nephropathyinduced renal fibrosis in rats,and to study the possible mechanism.METHODS:Sixty male Goto Kakizaki(GK) rats were randomly assigned to the model group,gliquidone group,astragaloside Ⅳ group,and high-,medium-and lowdoses DBD groups.After 8 weeks,changes in body weight,blood glucose,serum creatinine,serum urea nitrogen,and total cholesterol were observed.Changes in transforming growth factor-β1(TGF-β1),Smad3,and Smad5 pathways and the expression of the fibrosisrelated proteins collagen Ⅳ(col Ⅳ),α-smooth muscle actin(α-SMA),and vimentin were assessed.The degree of renal fibrosis was observed by immunohistochemistry and Mason staining.The expression of interleukin 6(IL-6),interleukin 10(IL-10),tumor necrosis factor(TNF-α),and C-reactive protein(CRP) in the kidneys was assessed using enzyme linked immunosorbent assay.RESULTS:Our experiments showed that DBD effectively reduced blood glucose,blood urea nitrogen,and creatinine levels after 8 weeks of administration,improved renal function in diabetic rats,alleviated renal fibrosis,and reduced the renal tissue levels of IL-6,IL-10,TNF-α,and CRP.Furthermore,DBD decreased the expression of TGF-β1,Smad3,col IV,α-SMA,and vimentin in renal tissues and increased the expression of Smad5.CONCLUSIONS:DBD ameliorates diabetic renal interstitial fibrosis by modulating the TGF-β1/Smads pathway.展开更多
OBJECTIVE:To explore the underlying molecular mechanism of Phellinus linteus(P.linteus).METHODS:We used a tandem mass tag-based quantitative proteomic method to determine the differentially expressed proteins.Network ...OBJECTIVE:To explore the underlying molecular mechanism of Phellinus linteus(P.linteus).METHODS:We used a tandem mass tag-based quantitative proteomic method to determine the differentially expressed proteins.Network pharmacology analysis was used to analysis the main components of P.linteus and construct the compound-target network.Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)were used to validate the analyses results.RESULTS:The expression levels of thrombospondin-1(TSP-1)and transforming growth factor(TGF)-β1/Smad3 signaling pathway proteins were significantly upregulated in focal segmental glomeruloscleosis(FSGS)rats.The P.linteus reduced the expression levels of TSP-1 and TGF-β1 signaling pathway proteins.Network pharmacology analysis revealed that protocatechualdehyde was the main active component.Subsequent in vivo and in vitro experiments validated the results of proteomic and network pharmacology analyses.CONCLUSIONS:Our results suggested that P.linteus may inhibit renal sclerosis by inhibiting TSP-1-activated TGF-β1 signaling and may have potential applications in the treatment of FSGS.展开更多
OBJEVTIVE:To investigate the efficacy of Cyclocarya paliurus(C.paliurus)polysaccharides on streptozotocin-induced diabetic nephropathy in rats.METHODS:Rats were divided into 6 groups,including group of normal control,...OBJEVTIVE:To investigate the efficacy of Cyclocarya paliurus(C.paliurus)polysaccharides on streptozotocin-induced diabetic nephropathy in rats.METHODS:Rats were divided into 6 groups,including group of normal control,group of diabetic control,group of metformin treatment,low-dose group of C.paliurus polysaccharides treatment,middle-dose group of C.paliurus polysaccharides treatment and high-dose group of C.paliurus polysaccharides treatment.Histological analysis of kidney was analyzed using hematoxilin and eosin.Levels of blood glucose,creatinine,urea,uric acid were determined by spectrophotometry.Anti-oxidative enzymes were measured by real-time polymerase chain reaction(PCR)and enzyme-linked immunosorbent assay(ELISA).Advanced glycation end products(AGEs)and transforming growth factor-β1(TGF-β1)level was measured by ELISA.RESULTS:Abnormal changes were observed in the group of diabetic control characterized by atrophy of the renal glomeruli with hypercellularity,congestion of glomerular tufts,dilation of the renal spaces,and degeneration of renal tubule.Compared with that of normal group,blood glucose,creatinine,urea,uric acid level was significantly increased in the group of diabetic control.Superoxide dismutase,catalase,glutathione peroxidase,glutathione reductase level was significantly decreased,but AGEs and TGF-β1 level was significantly increased.By contrast,administration of C.paliurus polysaccharides and metformin could reverse the above-mentioned results of the group of diabetic control,especially in the high-dose group of C.paliurus polysaccharides.CONCLUSION:Our findings suggest that C.paliurus polysaccharides may play a protecting role for nephropathy of diabetic rats by lowering glucose,creatinine,urea,uric acid level,enhancing the antioxidative ability,and reducing AGEs and TGF-β1 expression.展开更多
基金National Natural Science Foundation of China:a Metabolomic Study of the Effects of Moxibustion on Cardiac Function and Its Intervention in RA Model Rats Based on the TGF-β1/Smads Signaling Pathway (No.81403484)Anhui Province University Natural Science Fund Project of China:Exploring the Mechanism of Action of Moxibustion in AA Rats Based on Intestinal Flora and TLR4/NF-KB Signaling Pathway (No.KJ2019A0448)+2 种基金National Project Cultivation Fund Project Plan:Exploring the Mechanism of Action of Moxibustion in AA Rats Based on Intestinal Flora and TLR4/NF-KB Signaling Pathway (No.2019py01)Anhui Province Clinical Medical Research Center [Anhui Provincial Science and Technology Department Anhui Social Science (2020) No.41]the training Program of Outstanding talents in Colleges and Universities:2021 Domestic Visiting Training Program for Outstanding Young Key Teachers in Colleges and Universities (No.gxgnfx2021122)
文摘OBJECTIVE:To examine the effects of moxibustion on myocardial injury and myocardial metabolomics in rats with rheumatoid arthritis(RA)based on the transforming growth factor beta1(TGF-β1)/Smads signaling pathway.METHODS:One hundred rats were treated with saline[normal control(NC)group]or complete Freund’s adjuvant(CFA)by right plantar injection for the RA model group,and the latter were randomly divided into 4 groups.Tripterygium wilfordii polyglycoside tablets(雷公藤多苷片,TPT)have anti-inflammatory and are widely used in the clinical treatment of RA,therefore serving as a positive control group.Three days post injection rats were given TPT tablet(TPT group),acupuncture therapy(APT group),and moxibustion treatment(MOX group)for 15 consecutive days,while NC group and model group were equally grasped and fixed and received normal saline.Rat joint swelling scores and arthritis index(AI)were evaluated in each group before the CFA challenge,therapy and after receiving therapy.Myocardial ultrastructure was observed by electron microscope.Enzyme-linked immunosorbent assay was used to detect cardiac troponin I(cTnI)levels in rat myocardial tissue.Quantitative reverse transcription polymerase chain reaction and Western blotting analysis were used to measure the mRNA and protein levels of TGF-βsignaling molecules including TGF-β1,Smad2,Smad3,Smad4,and Smad7.Myocardial metabolomics was analyzed using gas chromatography-mass spectrometer.RESULTS:Compared with model group,RA model rats receiving TPT,acupuncture,or moxibustion therapy all showed reduced joint swelling scores and AI(all P<0.01)and improved myocardial damage,whereas rats treated with moxibustion were found to be more marked.Consistently,the expressions of cTnI,TGF-β1,Smad2,Smad3,and Smad4 were found to be elevated in model rat group in contrast to NC rats and were significantly downregulated in TPT,APT and MOX group when compared with model group,while the levels of Smad7 showed the opposite result(all P<0.01).Moreover,the dissection of metabolomics suggested a novel metabolite biomarker panel including D-Xylulose 5-phosphate,dihydroxyacetone phosphate,arachidonic acid,etc was defined and implicated in amino acid,glucose,and fatty acid metabolic processes as revealed by principal component analysis and partial least squares discriminant analysis.CONCLUSION:Moxibustion prevents RA-induced inflammatory response and offers potent therapeutic effects on myocardial dysfunctions.The protective effects might be associated with its role in TGF-β1 inactivation and metabolic reprogramming.
基金National Natural Science Foundation of China:Based on TGF-β1/Smads Signaling Pathway to Study the Effect Mechanism of Tuina along the Bladder Meridian on Intervertebral Disc Degeneration(82004497)China Postdoctoral Science Foundation:Based on TGF-β1/RhoA/JNK Signaling Pathway to Study the Effect Mechanism of Tuina along the Bladder Meridian on Intervertebral Disc Degeneration(No.2021M693788)。
文摘OBJECTIVE:The aim of this study was to investigate the protective effects of Tuina(a traditional Chinese massage therapy)on intervertebral disc(IVD)degeneration and the regulatory mechanisms of the transforming growth factor-β1(TGF-β1)/small mothers against decapentaplegic(Smad)signaling pathway.METHODS:Thirty New Zealand white rabbits were randomized into five groups:the control group,model group,model+Tuina group(Tuina group),model+TGF-β1 group(TGF-β1 group),and model+TGF-β1 inhibitor SB431542 group(SB431542 group).The model was established by posterolateral annulus fibrosus puncturing(AFP).Recombinant TGF-β1 and inhibitor SB431542 was injected into the TGF-β1 group and SB431542 group with a microsyringe,respectively.The rabbits in the Tuina group received Tuina treatment along the bladder meridian for 4 weeks.Magnetic resonance imaging(MRI)was performed on rabbits before AFP and after 4 weeks of intervention.Lumbar IVDs(L2-L3 to L4-L5)were harvested after intervention.Histopathological changes in the IVDs were measured by hematoxylin and eosin(HE)staining.Type I collagen was analyzed by immunohistochemistry detection.The expression level of matrix metalloproteinase-3(MMP3)was determined by enzyme-linked immunosorbent assay.Cell apoptosis was evaluated by terminal deoxynucleotidyl transferasemediated nick end labeling and Western blotting.Realtime polymerase chain reaction and Western blotting were used to analyze the expression of TGF-β1 and Smad2/3/4 and a disintegrin and metalloproteinase with thrombospondin motifs 5.RESULTS:Posterolateral AFP induced IVD degeneration in rabbits with histopathological damage and noticeable changes in MRI images.Tuina alleviated histopathological changes and reversed the expression of extracellular matrix degeneration-related molecules and apoptosis-related proteins.Furthermore,AFP induced the activation of TGF-β1 and Smad2/3/4,whereas Tuina therapy markedly reduced the protein expression of Smad2/3 and the gene expression of TGF-β1 and Smad2/3/4.Additionally,the TGF-β1/Smad signaling pathway was activated in the TGF-β1 group,while the TGF-β1/Smad signaling pathway was inhibited in the SB431542 group.CONCLUSION:Posterolateral AFP induced disc degeneration as determined by MRI assessment and histological analysis.Tuina alleviated disc degeneration,possibly by inhibiting the fibrotic response mediated by the TGF-β1/Smad pathway,thus alleviating extracellular matrix degeneration and reducing cell apoptosis.
基金Supported by the Scientific Research Project of Heilongjiang Administration of Traditional Chinese Medicine(ZHY18-087):Clinical Study on the Treatment of Tubal Inflammatory Infertility with Gexia Zhuyu Decoction Combined with Hysteroscopic Tubal Fluid PassageNatural Science Foundation of Heilongjiang Province(H2015026):Study on the Effect of Activating Blood Circulation and Removing Blood Stasis,Activating Qi and Relieving Pain on Salpingitis Infertile Rat ModelMechanism of Gexia Zhuyu Decoction Improved Fallopian Tube Fibrosis in the Treatment of Salpingitic Obstructive Infertility based on Transforming Growth Factor-Β1/Phosphorylation38 Mitogen-Activated Protein Kinase Pathway(2021ZYGLG001)。
文摘OBJECTIVE:To explore the effect of electroacupuncture(EA)on rats with chronic fallopian tube inflammation and its potential mechanisms.METHODS:Thirty-six female Sprague-Dawley rats were divided into Control,Model and EA groups.The pathological morphology of the fallopian tubes was observed by hematoxylin-eosin(HE)and Masson staining.The results of transforming growth factor-β1(TGF-β1),P38 mitogen-activated protein kinase(MAPK),phosphorylation(p)-p38MAPK in rat oviduct tissues were detected by immunohistochemistry.Results of P38MAPK,p-P38MAPK and TGF-β1 in rat oviduct tissues were detected by immunofluorescence.The expression level of p38MAPK,p-P38MAPK,TGF-β1 protein in rats was detected by Western blot.Quantitative real-time polymerase chain reaction(RT-q PCR)was used to detect m RNA expression levels of TGF-β1.RESULTS:It found that collagen fibers counts decreased significantly in EA group compared to Model group.The phosphorylation of P38MAPK in EA group was significantly reduced compared to Model group.The serum TGF-β1 expressions in EA group increased decreased significantly.CONCLUSION:Electroacupuncture was able to attenuate chronic salpingitis through down-regulating TGF-β1/MAPK signaling pathway.
基金Supported by the Department of Science and Technology of Southwest Medical University(No.2017-ZRQN-072)the Department of Science and Technology of Affiliated Hospital of Southwest Medical University(No.16231)。
文摘OBJECTIVE:To investigate the efficacy of Shenweifang(SWF)-containing serum on transforming growth factor(TGF)-β1–induced fibroblast-myofibroblast transition in normal rat kidney interstitial fibroblast cells(NRK-49 F).METHODS:Sprague-Dawley rats were gavaged with one of five solutions:(a)saline;(b)saline plus low-dose SWF;(c)saline plus medium-dose SWF;(d)saline plus highdose SWF;and(e)saline plus valsartan.NRK-49 F cells were treated with TGF-β1 and cultured using serum from the gavaged rats.RESULTS:TGF-β1 treatment increased the expression ofα-smooth muscle actin,proliferating cell nuclear antigen,collagenⅠ,Smad3,mitogen-activated protein kinase(MAPK)10,and c-Jun N-terminal kinase(JNK)3 and induced abnormalities in cell morphology,cell cycle progression,and cell proliferation.CONCLUSIONS:SWF-or valsartan-containing serum corrected(or partially corrected)TGF-β1–induced abnormal changes in this in vitro system.SWF-containing serum reversed abnormalities in morphology,cell cycle progression,and proliferation in TGF-β1–treated NRK-49F cells,probably by blocking the TGF-β1/Smads and TGF-β1/MAPK/JNK pathways.
基金Sponsored by the National Natural Science Fund (No. 81903994)Youth Development of the First Affiliated Hospital of Anhui Medical University (No. 2793)the Budget Project of Shanghai University of Chinese Medicine (No. 2019LK095)。
文摘OBJECTIVE: To investigate the effect of Huangqi decoction( 黄芪汤) on renal interstitial fibrosis and its association with the transforming growth factor-β1(TGF-β1)/mitogen-activated protein kinase(MAPK) signaling pathway. METHODS: 120 C57/BL mice were randomly divided into six groups: sham group, Enalapril(20 mg/kg) group, 5/6 nephrectomy model group, and 5/6 nephrectomy model plus Huangqicoction(0.12, 0.36 and 1.08 g/kg respectively) groups. Detecting 24hours urinary protein, blood pressure, serum creatinine, urea nitrogen content changes. Periodic Acid-Schiff stain(PAS) and Masson’s trichrome staining was used to observe the renal tissue pathological changes. Protein expression of TGF-β1, Phosphorylated P38 mitogen activated protein kinases(P-P38), Phosphorylated c-jun N-terminal kinase(P-JNK), Phosphorylated extracellular regulated proteinhnase(PERK), Fibroblast-specific protein-1(FSP-1), Alpha smooth muscle actin(α-SMA), Type Ⅲ collagen(Collagen Ⅲ), Connective tissue growth factor(CTGF),Bcl-2 Assaciated X protein(Bax) and B cell lymphoma 2(Bcl-2) were measured with western blot and immunohistochemical. RESULTS: Both Huangqi decoction and Enalapril improved the kidney function, 24 h urinary protein and the fibrosis in 5/6 nephrectomy mice, Huangqi decoction downregulated the expressions of TGF-β1, FSP-1, α-SMA, Collagen Ⅲ and CTGF in a dose-dependent manner, and it has a significant difference(P < 0.01) compared with model group.Huangqi decoction downregulated the expressions of P-P38, P-JNK, P-ERK and Bcl-2 in a dose-dependent manner, while upregulated the expression of Bax. CONCLUSIONS: The protective effect of Huangqi decoction for renal interstitial fibrosis in 5/6 nephrectomized mice via the inhibition of EpithelialMesenchymal Transitions and downregulating the TGF-β1/MAPK signaling pathway.
基金Supported by the Government Funded Clinical Medicine Eexcellent Talent Training and Basic Research Project PlanNatural Science Foundation of Hebei(No.H2019423092)+2 种基金Higher Education Science and Technology Research Project of Hebei(No.ZD2016056)Postgraduate Innovation Ability Development Project of Hebei Education Department(No.CXZZBS2019159)Basic Research Business Expenses of Provincial Universities of Hebei University of Chinese Medicine Project of Excellent Student Research Capacity Improvement(No.YXZ2019001)。
文摘OBJECTIVE: To investigate the effect of Danggui Buxue Tang(DBT), a decoction from Traditional Chinese Medicine, on bleomycin-induced pulmonary fibrosis in rats, and to propose the possible underlying mechanism.METHODS: Forty male Sprague-Dawley rats were randomly divided into sham group, model group,prednisone group and DBT group. Pulmonary fibrosis rat model was established by intratracheal injection with bleomycin. Body weight and lung index were monitored. Histopathologic examination and collagen deposition were determined using Hematoxylin and eosin(HE) and Masson's trichrome staining. Immunohistochemistry staining was applied to observe the expression of alpha-smooth muscle actin(α-SMA). m RNA expression of α-SMA,collagen Ⅰ and collagen Ⅲ were measured by realtime fluorescence quantitative PCR(RT-q PCR). Inflammatory cytokines, including tumor necrosis factor alpha(TNF-α), interleukin-6(IL-6) and IL-1β in serum were detected by Enzyme-linked immunosorbent assay. Alkali hydrolysis method was conducted to investigate the content of hydroxyproline(HYP). Transforming growth factor-β1(TGF-β1),Smad3 and plasminogen activator inhibitor-1(PAI-1) protein level were examined by Western blot assay.RESULTS: DBT significantly reduced the severity of bleomycin-induced pulmonary fibrosis and inflammation as indicated by minimizing the lost of weight, and by lowering the levels of lung index, inflammation score, Ashcroft score, collagen volume fraction(%), HYP, α-SMA, collagen Ⅰ, collagen Ⅲ,TNF-α, IL-6, IL-1β, TGF-β1, Smad3 and PAI-1, consistent with the effect of prednisone.CONCLUSION: Our findings suggest that DBT is able to ameliorate the pulmonary fibrosis, the possible mechanism may involve inhibition of pulmonary inflammation and collagen deposition, possibly via suppressing TGF-β1/Smad3/PAI-1 signaling pathway.
基金Supported by the National Natural Science Foundation of China(Inducing effect of Qingguang'an on Autophagy of Tenon's Capsule Fibroblasts after Glaucoma Filtration Surgery,No.81603665)And the Postdoctoral Science Foundation of China(Experimental Study on Autophagy of Tenon's Fibroblasts Induced by Qingguang'an after Glaucoma Filtration Surgery,No.2017M612565)。
文摘OBJECTIVE:To explore the effects of Qingguang'an(青光安)containing serum on the expression levels of autophagy related genes in the transforming growth factor beta 1(TGF-β1)-activated human Tenon's fibroblasts(HTFs).METHODS:(a)Primary HTFs were stimulated by TGF-β1 and underwent immunohistochemistry,which established a cell model after Glaucoma filtration surgery(GFS).(b)The cell models were divided into 4 group:normal group(normal cells),model group(+TGF-β1),treatment group(+TGF-β1+medicated serum),and positive control group(TGF-β1+rapamycin).Then,Qingguang'an medicated serum with optimum concentration was added to the corresponding group.The autophagy positive cells were identified by the Cyto-ID autophagy detection kits under fluorescent microscope and Cytation 5 multifunctional instrument for cell imaging.And the mean fluorescence intensity of autophagy positive cells was determined by flow cytometry.The expression levels of autophagy related genes—Beclin-1,autophagy related gene 5(ATG-5),and microtubule-associated protein 1 light chain 3(LC-3Ⅱ)were detected by quantitative reverse transcription-polymerase chain reaction and Western blot analysis.RESULTS:Compared with the normal group and the model group,the relative mRNA expression levels of autophagy-related genes(Beclin-1,ATG-5 and LC-3Ⅱ)in the experimental group were notably increased(P<0.05,P<0.01),and with the extension of treatment time,it had an increasing trend(48 h was more obvious),which showed a certain time dependency;the protein expression levels of autophagy-related genes(Beclin-1,ATG-5,and LC-3Ⅱ)were significantly increased in the experimental group(P<0.05,P<0.01).With the prolongation of treatment time,there was an increasing trend(48 h was relatively obvious),and it revealed a certain time dependency CONCLUSION:The Qingguang'an medicated serum could up-regulate autophagy related genes(Beclin1,ATG5,and LC3Ⅱ)in the TGF-β1-activated HTFs.
基金financially supported by the Hamadan University of Medical Sciences(No 9603302213)
文摘Objective:To evaluate the effect of resveratrol against CCl4-induced nephrotoxicity.Methods:Forty-two male Wistar rats were divided into seven groups randomly.After six weeks,kidney weight,body weight,blood urea,serum creatinine,oxidative stress markers,and gene expression of renal transforming growth factor-beta1(TGF-β1),TGF-βreceptor type 1(TGF-βR1)and Smad3 were determined.In addition,the protein level of TGF-β1 in the tissue lysate was measured.Results:Resveratrol had a protective role in renal tissue by the improvement of antioxidant balance and reduction of renal parameters such as creatinine and urea(P<0.001).In addition,the renal m RNA level of TGF-β1,TGF-βR1,Smad3,as well as the protein level of TGF-β1 were decreased in rats treated with resveratrol(P<0.001),compared to the CCl4 group.Conclusions:Overall,resveratrol shows a protective effect against nephrotoxicity in CCl4 treated rats by reducing oxidative stress status and modulating the TGF-βsignaling.
基金supported by the National Natural Science Foundation of China,No.81801175(to CLT)the Fundamental Research Funds for the Central Universities of China,No.WK9110000044(to CLT)+2 种基金China Scholarship Council,No.201706270155(to CLT)the China Postdoctoral Science Foundation,No.2019M662179(to CLT)the Anhui Province Postdoctoral Science Foundation of China,No.2019B324(to CLT)
文摘Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,and its dynamic changes are closely related to the proliferation of endothelial cells,the recruitment of pericytes to blood vessels,and functional differentiation during embryonic vascular development.The pathophysiology of many cerebrovascular diseases is today understood as a disorder of endothelial cell function and an imbalance in the proportion of vascular cells.Indeed,mutations in ALK1 and its co-receptor endoglin are major genetic risk factors for vascular arteriovenous malformation.Many studies have shown that ALK1 is closely related to the development of cerebral aneurysms,arteriovenous malformations,and cerebral atherosclerosis.In this review,we describe the various roles of ALK1 in the regulation of angiogenesis and in the maintenance of cerebral vascular homeostasis,and we discuss its relationship to functional dysregulation in cerebrovascular diseases.This review should provide new perspectives for basic research on cerebrovascular diseases and offer more effective targets and strategies for clinical diagnosis,treatment,and prevention.
文摘Stress, inflammation and Plasminogen activator inhibitor 1 (PAI-1) are key mechanisms throughout the development of aging, constituting a crossroad in the most frequent pathologies that accompany it. Among metabolic processes, obesity, metabolic syndrome and type 2 diabetes mellitus are included and Alzheimer’s disease among the neurodegenerative processes. Stress is a mechanism of defense of the organism against exogenous and endogenous actions called stressors. In the case of low intensity stimuli, the organism responds with actions aimed at a physiological adaptation (Homeostasis). On the other hand, when a high intensity (experimental level) or chronic stimulus (oxidative stress) is repeated, structural and functional changes are observed in different organs with activation of the hypothalamus-pituitary-adrenal axis, the renin angiotensin system and the sympathetic nervous system, stimulating the production of hormones that release cytokines with proin-flammatory/antiinflammatory properties that play an important role in the previously mentioned pathologies, as well as a marked increase in PAI-1, a gene regulated by stress and by cytokines, with manifest action at the origin of thromboembolic disease, so frequent in aging. The objective of this review is to highlight the importance of the binomial stress and PAI-1 in aging and in the pathologies that accompany it. Because PAI-1 is part of the pathology and complications in aging, some authors suggest the study of PAI-1 inhibitors to achieve its physiological levels, as part of the treatment of these diseases.
基金Supported by National Natural Science Foundation of China Youth Fund:Research on the Mechanism of Danggui Buxue Decoction in the Treatment of Diabetic Renal Interstitial Fibrosis Based on the Regulation of TGF-β1/Smad3 Signaling Pathway by Mi RNA-27a(No.81904085)Nanjing University of Chinese Medicine Foundation Youth Project:Research on the Mechanism of Danggui Buxue Decoction in the Treatment of Diabetic Renal Interstitial Fibrosis Based on the Regulation of TGF-β1/Smad3 Signaling Pathway by Mi RNA-27a(No.NZY81904085)。
文摘OBJECTIVE:To observe the efficacy of Danggui Buxue decoction(当归补血汤,DBD) on diabetic nephropathyinduced renal fibrosis in rats,and to study the possible mechanism.METHODS:Sixty male Goto Kakizaki(GK) rats were randomly assigned to the model group,gliquidone group,astragaloside Ⅳ group,and high-,medium-and lowdoses DBD groups.After 8 weeks,changes in body weight,blood glucose,serum creatinine,serum urea nitrogen,and total cholesterol were observed.Changes in transforming growth factor-β1(TGF-β1),Smad3,and Smad5 pathways and the expression of the fibrosisrelated proteins collagen Ⅳ(col Ⅳ),α-smooth muscle actin(α-SMA),and vimentin were assessed.The degree of renal fibrosis was observed by immunohistochemistry and Mason staining.The expression of interleukin 6(IL-6),interleukin 10(IL-10),tumor necrosis factor(TNF-α),and C-reactive protein(CRP) in the kidneys was assessed using enzyme linked immunosorbent assay.RESULTS:Our experiments showed that DBD effectively reduced blood glucose,blood urea nitrogen,and creatinine levels after 8 weeks of administration,improved renal function in diabetic rats,alleviated renal fibrosis,and reduced the renal tissue levels of IL-6,IL-10,TNF-α,and CRP.Furthermore,DBD decreased the expression of TGF-β1,Smad3,col IV,α-SMA,and vimentin in renal tissues and increased the expression of Smad5.CONCLUSIONS:DBD ameliorates diabetic renal interstitial fibrosis by modulating the TGF-β1/Smads pathway.
基金Public Welfare Technology Application Research Program of Zhejiang Province:Study on the Application of Phellinus linteus Protocatechualdehyde in Inhibiting Glomerular Extracellular Matrix Accumulation based on TSP-1 Double Luciferase Reporting System(No.LGC21H290002)Key Projects of Zhejiang Administration of Traditional Chinese Medicine:Quality and Safety Evaluation of Cultivated Phellinus linteus in Zhejiang Province and its Comprehensive Application in the Prevention and Treatment of Kidney Disease(No.2020ZZ016)。
文摘OBJECTIVE:To explore the underlying molecular mechanism of Phellinus linteus(P.linteus).METHODS:We used a tandem mass tag-based quantitative proteomic method to determine the differentially expressed proteins.Network pharmacology analysis was used to analysis the main components of P.linteus and construct the compound-target network.Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)were used to validate the analyses results.RESULTS:The expression levels of thrombospondin-1(TSP-1)and transforming growth factor(TGF)-β1/Smad3 signaling pathway proteins were significantly upregulated in focal segmental glomeruloscleosis(FSGS)rats.The P.linteus reduced the expression levels of TSP-1 and TGF-β1 signaling pathway proteins.Network pharmacology analysis revealed that protocatechualdehyde was the main active component.Subsequent in vivo and in vitro experiments validated the results of proteomic and network pharmacology analyses.CONCLUSIONS:Our results suggested that P.linteus may inhibit renal sclerosis by inhibiting TSP-1-activated TGF-β1 signaling and may have potential applications in the treatment of FSGS.
基金Supported by the by the Natural Science Foundation of Henan Effect of Cyclocarya paliurus polysaccharides on the diabetes mellitus(No.182300410123)。
文摘OBJEVTIVE:To investigate the efficacy of Cyclocarya paliurus(C.paliurus)polysaccharides on streptozotocin-induced diabetic nephropathy in rats.METHODS:Rats were divided into 6 groups,including group of normal control,group of diabetic control,group of metformin treatment,low-dose group of C.paliurus polysaccharides treatment,middle-dose group of C.paliurus polysaccharides treatment and high-dose group of C.paliurus polysaccharides treatment.Histological analysis of kidney was analyzed using hematoxilin and eosin.Levels of blood glucose,creatinine,urea,uric acid were determined by spectrophotometry.Anti-oxidative enzymes were measured by real-time polymerase chain reaction(PCR)and enzyme-linked immunosorbent assay(ELISA).Advanced glycation end products(AGEs)and transforming growth factor-β1(TGF-β1)level was measured by ELISA.RESULTS:Abnormal changes were observed in the group of diabetic control characterized by atrophy of the renal glomeruli with hypercellularity,congestion of glomerular tufts,dilation of the renal spaces,and degeneration of renal tubule.Compared with that of normal group,blood glucose,creatinine,urea,uric acid level was significantly increased in the group of diabetic control.Superoxide dismutase,catalase,glutathione peroxidase,glutathione reductase level was significantly decreased,but AGEs and TGF-β1 level was significantly increased.By contrast,administration of C.paliurus polysaccharides and metformin could reverse the above-mentioned results of the group of diabetic control,especially in the high-dose group of C.paliurus polysaccharides.CONCLUSION:Our findings suggest that C.paliurus polysaccharides may play a protecting role for nephropathy of diabetic rats by lowering glucose,creatinine,urea,uric acid level,enhancing the antioxidative ability,and reducing AGEs and TGF-β1 expression.
