AIM： To determine if tranilast affects human corneal fibroblast（HCFs） contraction.METHODS： HCFs cultured in a three-dimensional type I collagen gel were treated with or without transforming growth factor beta（TGF...AIM： To determine if tranilast affects human corneal fibroblast（HCFs） contraction.METHODS： HCFs cultured in a three-dimensional type I collagen gel were treated with or without transforming growth factor beta（TGF-β） or tranilast. Gel diameter was measured as an indicator for collagen contraction. Immunoblot was performed to evaluate myosin light chain（MLC） and paxillin phosphorylation. Confocal microscopy was employed to examine the focal adhesions and actin stress fiber formation. Immunoblot analysis and gelatin zymography were performed to detect tissue inhibitors of metalloproteinases and matrix metalloproteinases（MMPs） in supernatant.RESULTS： The inhibitory effect of tranilast on HCFsmediated collagen gel contraction induced by TGF-β was dose-dependent. The significant effect of tranilast was started from 100 μmol/L and maximized at 300 μmol/L. The peak effect of 300 μmol/L tranilast also relied on the duration of treatment, which showed statistical significance from day 2. TGF-β-induced paxillin and MLC phosphorylation, stress fiber formation, focal adhesions, and MMP-1, MMP-2, and MMP-3 secretion in HCFs were also inhibited by tranilast.CONCLUSION： Tranilast suppresses the HCFs-cultured collagen gel contraction induced by TGF-β. It attenuates actin stress fibers formation, focal adhesions, and the secretion of MMPs, with these actions likely contributing to the inhibitory effect on HCF contractility. By attenuating the contractility of corneal fibroblasts, tranilast treatment may inhibit corneal scarring.展开更多
Objective:Cold exposure is associated with increased prevalence of hypertension and the related severe cardiovascular events.Aberrant activation of the nucleotide-binding oligomerization domain-like receptor protein 3...Objective:Cold exposure is associated with increased prevalence of hypertension and the related severe cardiovascular events.Aberrant activation of the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome plays an important role in the development of hypertension.Tranilast(TR),an inhibitor of NLRP3,provides a useful pharmacological probe for exploring the role of NLRP3 in pathogenesis associated with inflammation and its potential application as a therapeutic agent.This study was designed to examine the effects of TR on NLRP3 and hypertension in rats exposed to cold environment to simulate the frigid-zone conditions.Methods and results:Sprague Dawley(SD)rats were exposed to moderate cold temperature(4±1℃),and then were randomized to receive TR or vehicle for 3 weeks,while the control group was raised under rat room temperature(RT,23±1℃).We found that cold exposure substantially increased blood pressure,NLRP3 inflammasome level,and fibrosis in aorta,which were reversed by TR.Conclusion:TR has an anti-hypertensive property in cold environment,and this beneficial action is likely conferred by its inhibitory effects on inflammation and fibrosis.These findings suggest TR as a potential drug for the treatment of cold-induced hypertension.展开更多
基金Supported by the National Science Foundation of China (No.81770889)the Natural Science Foundation of Guangdong Province (No.2017A030313774)the Research Fund of Jilin Provincial Science and Technology Department to Yang Liu (International Cooperation Item, No.20160414055GH)
文摘AIM： To determine if tranilast affects human corneal fibroblast（HCFs） contraction.METHODS： HCFs cultured in a three-dimensional type I collagen gel were treated with or without transforming growth factor beta（TGF-β） or tranilast. Gel diameter was measured as an indicator for collagen contraction. Immunoblot was performed to evaluate myosin light chain（MLC） and paxillin phosphorylation. Confocal microscopy was employed to examine the focal adhesions and actin stress fiber formation. Immunoblot analysis and gelatin zymography were performed to detect tissue inhibitors of metalloproteinases and matrix metalloproteinases（MMPs） in supernatant.RESULTS： The inhibitory effect of tranilast on HCFsmediated collagen gel contraction induced by TGF-β was dose-dependent. The significant effect of tranilast was started from 100 μmol/L and maximized at 300 μmol/L. The peak effect of 300 μmol/L tranilast also relied on the duration of treatment, which showed statistical significance from day 2. TGF-β-induced paxillin and MLC phosphorylation, stress fiber formation, focal adhesions, and MMP-1, MMP-2, and MMP-3 secretion in HCFs were also inhibited by tranilast.CONCLUSION： Tranilast suppresses the HCFs-cultured collagen gel contraction induced by TGF-β. It attenuates actin stress fibers formation, focal adhesions, and the secretion of MMPs, with these actions likely contributing to the inhibitory effect on HCF contractility. By attenuating the contractility of corneal fibroblasts, tranilast treatment may inhibit corneal scarring.
文摘Objective:Cold exposure is associated with increased prevalence of hypertension and the related severe cardiovascular events.Aberrant activation of the nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome plays an important role in the development of hypertension.Tranilast(TR),an inhibitor of NLRP3,provides a useful pharmacological probe for exploring the role of NLRP3 in pathogenesis associated with inflammation and its potential application as a therapeutic agent.This study was designed to examine the effects of TR on NLRP3 and hypertension in rats exposed to cold environment to simulate the frigid-zone conditions.Methods and results:Sprague Dawley(SD)rats were exposed to moderate cold temperature(4±1℃),and then were randomized to receive TR or vehicle for 3 weeks,while the control group was raised under rat room temperature(RT,23±1℃).We found that cold exposure substantially increased blood pressure,NLRP3 inflammasome level,and fibrosis in aorta,which were reversed by TR.Conclusion:TR has an anti-hypertensive property in cold environment,and this beneficial action is likely conferred by its inhibitory effects on inflammation and fibrosis.These findings suggest TR as a potential drug for the treatment of cold-induced hypertension.
