17β-estradiol inhibits TGF-β-induced collagen gel contraction mediated by human Tenon fibroblasts via Smads and MAPK signaling pathways  

作　　者：Cheng-Cheng Yang Meng-Jie Liu Yun-Ze-Peng Li Zheng-Hua Xu Yang Liu Zi-Han Guo Bin-Hui Li Xiu-Xia Yang 

机构地区：[1]Department of Ophthalmology,the Fifth Affiliated Hospital,Sun Yat-sen University,Zhuhai 519000,Guangdong Province,China [2]Eye Institute of Xiamen University,Fujian Provincial Key Laboratory of Ophthalmology and Visual Science,School of Medicine,Xiamen University,Xiamen 361000,Fujian Province,China

出　　处：《International Journal of Ophthalmology(English edition)》2023年第9期1441-1449,共9页国际眼科杂志（英文版）

基　　金：Supported by the National Natural Science Foundation of China(No.81770889);Zhuhai Science and Technology Program(No.ZH22036201210134PWC).

摘　　要：AIM:To investigate the impact of 17β-estradiol on the collagen gels contraction(CGC)and inflammation induced by transforming growth factor(TGF)-βin human Tenon fibroblasts(HTFs).METHODS:HTFs were three-dimensionally cultivated in type I collagen-generated gels with or without TGF-β(5 ng/mL),17β-estradiol(12.5 to 100μmol/L),or progesterone(12.5 to 100μmol/L).Then,the collagen gel diameter was determined to assess the contraction,and the development of stress fibers was analyzed using immunofluorescence staining.Immunoblot and gelatin zymography assays were used to analyze matrix metalloproteinases(MMPs)and tissue inhibitors of metalloproteinases(TIMPs)being released into culture supernatants.Enzyme-linked immunosorbent assay(ELISA)and reverse transcription-quantitative polymerase chain reaction(RT-PCR)were used to detect interleukin(IL)-6,monocyte chemoattractant proteins(MCP)-1,and vascular endothelial growth factor(VEGF)in HTFs at the translational and transcriptional levels.The phosphorylation levels of Sma-and Mad-related proteins(Smads),mitogen-activated protein kinases(MAPKs),and protein kinase B(AKT)were measured by immunoblotting.Statistical analysis was performed using either the Tukey-Kramer test or Student’s unpaired t-test to compare the various treatments.RESULTS:The CGC caused by TGF-βin HTFs was significantly inhibited by 17β-estradiol(25 to 100μmol/L),and a statistically significant difference was observed when comparing the normal control group with 17β-estradiol concentrations exceeding 25μmol/L(P<0.05).The suppressive impact of 17β-estradiol became evident 24h after administration and peaked at 72h(P<0.05),whereas progesterone had no impact.Moreover,17β-estradiol attenuated the formation of stress fibers,and the production of MMP-3 and MMP-1 in HTFs stimulated by TGF-β.The expression of MCP-1,IL-6,and VEGF mRNA and protein in HTFs were suppressed by 100μmol/L 17β-estradiol(P<0.01).Additionally,the phosphorylation of Smad2 Smad3,p38,and extracellular signal-regulated kinase(ERK)

关 键 词：Tenon fibroblasts transforming growth factor-β 17Β-ESTRADIOL FIBROSIS wound healing 

分 类 号：R777.3[医药卫生—眼科]