眼科新进展2018，Vol.38Issue(4)：319-323,5.DOI:10.13389/j.cnki.rao.2018.0075
杞精明目汤颗粒剂对肿瘤坏死因子-α刺激下结膜松弛症成纤维细胞MAPK信号通路的影响
Effect of Qijingmingmu decoction granule on MAPK signal pathway in fibroblasts of conjunctivochalasis stimulated by TNF-α
摘要
Abstract
Objective To observe the effects of Qijingmingmu decoction granule on the expression of mitogen-activated protein linase (MAPK) signal pathway in fibroblasts of conjunctivochalasis (CCH) under the stimulation of tumor necrosis factor (TNF-α) and to explore the pathogenesis and effective treatment method of CCH.Methods CCH conjunctival fibroblasts were cultured in vitro and divided into CCH group,CCH + TNF-α group and CCH + TNF-α + Qijingmingmu decoction group.CCK-8 assay was used to determine the effective concentration of Qijingmingmu decoction granule,and 10-2 mg · L-1 TNF-α was added in the cultured fibroblasts of the latter two groups,followed by interference with Qijingmingmu decoction granule for 48h.The expression of MAPK signal pathway related protein and mRNA were detected by ELISA,Western Blot and RT-PCR.Then the results were statistically analyzed.Results CCK-8 assay showed the effective concentration of Qijingmingmu decoction granule was 1.61g · L-1.And there were significant differences in the total A values (A450) of extracellular regulated protein kinase (ERK),c-jun N-terminal kinase (JNK),p38 mitogen activated protein kinase (p38 MAPK) and their phosphorylation levels among the three groups (all P ＜ 0.05).Moreover,TNF-α could significantly up-regulate the expression of ERK1/2,JNK1/2,p-JNK1/2,p38 MAPK and p-p38 MAPK in CCH fibroblasts (all P ＜0.05),while Qijingmingmu decoction down-regulated their expressions in CCK fibroblasts after TNF-α stimulation (all P ＜ 0.05).Furthermore,the total differences in p-ERK1/2,p38 MAPK and p-p38 MAPK protein were significant in the three groups (all P＜0.05);and TNF-α could significantly up-regulate the expression of p-ERK1/2,p-JNK1/2,p38 MAPK and p-p38 MAPK protein in CCH fibroblasts (all P ＜0.05),while Qijingmingmu decoction down-regulated p-ERK1/2 and p38 MAPK expressions in CCK fibroblasts after TNF-α stimulation (both P ＜ 0.05).In addition,the total differences in ERK1/2 and p38 MAPK mRNA were significant in the three groups (both P ＜ 0.05);and TNF-α could significantly up-regulate the expression of ERK1/2 and p38 MAPK mRNA in CCH fibroblasts (both P ＜ 0.05),while Qijingmingmu decoction down-regulated p38 MAPK mRNA expressions in CCK fibroblasts after TNF-α stimulation (P ＜ 0.05).Conclusion The inflammatory factor TNF-α can up-regulate the expressions of MAPK signal pathway related protein and mRNA in CCH fibroblasts,resulting in the occurrence and development of CCH,and meanwhile Qijingmingmu decoction granule can downregulate the expression of MAPK signal pathway to play the therapeutic role in CCH to some extent.关键词
结膜松弛症/成纤维细胞/肿瘤坏死因子α/丝裂原活化蛋白激酶/杞精明目汤颗粒剂Key words
conjunctivochalasis/fibroblast/TNF-α/mitogen-activated protein kinase/Qijingmingmu decoction granule分类
医药卫生引用本文复制引用
贾元玲,项敏泓,文杭,李青松,詹月萍,黄丽,刘晓静..杞精明目汤颗粒剂对肿瘤坏死因子-α刺激下结膜松弛症成纤维细胞MAPK信号通路的影响[J].眼科新进展,2018,38(4):319-323,5.基金项目
上海中医药大学预算内项目(编号:2015YSN50) （编号:2015YSN50）
上海市医学重点专科项目(编号:ZK2015A20) （编号:ZK2015A20）
上海市普陀区中心医院英才计划项目(编号:2017202B)Shanghai University of Traditional Chinese Medicine Budget Project (No:2015YSN50) （编号:2017202B）
Shanghai Key Medical Specialist-Department of Ophthalmology (No:ZK2015A20) （No:ZK2015A20）
Elite Plan of Shanghai Central Hospital of Putuo District (No:2017202B) （No:2017202B）
