摘要
目的:探讨金乌健骨方对类风湿关节炎(RA)滑膜细胞自噬基因LC3、Beclin-1、Bcl-2、VPS34表达的影响。方法:取3代RA滑膜细胞,加金乌健骨方总提物低、中、高浓度(0.06、0.6、6.0 mg/mL)及雷公藤多苷(0.03 mg/mL)干预24 h后,采用透射电镜观察细胞自噬情况,PCR检测LC3、Beclin-1、Bcl-2、VPS34 mRNA表达水平;Western blot检测LC3、Beclin-1蛋白表达。结果:空白对照组细胞无明显自噬小体或自噬溶酶体,各给药组滑膜细胞胞浆中自噬小体和自噬溶酶体明显增多。与空白对照组比较,除金乌健骨方低浓度组外,各给药组滑膜细胞LC3、Beclin-1、VPS34 mRNA表达均显著降低,Bcl-2 mRNA表达显著升高,LC3、Beclin-1蛋白表达显著降低(P<0.01)。以金乌健骨方高浓度组和雷公藤多苷组作用最明显。结论:苗药金乌健骨方能抑制RA滑膜细胞自噬相关基因LC3、Beclin-1、VPS34的表达,增强Bcl-2的表达水平。
Objective:To investigate the effect of Jinwu jiangu decoction on the expressions of autophagy genes LC3,Beclin-1,Bcl-2 and VPS34 in synovial cells of rheumatoid arthritis(RA).Methods:After 24 h of intervention with the low,medium,high concentration(0.06,0.6,6.0 mg/mL)of total extract of Jinwu jiangu decoction and tripterygium glycoside(0.03 mg/mL),the autophagy of RA synovial cells was observed by transmission electron microscopy.The expression levels of LC3,Beclin-1,Bcl-2 and VPS34 mRNA were detected by PCR,and the protein expressions of LC3 and Beclin-1 were detected by Western blot.Results:There was no significant autophagic bodies or lysosomes in the blank control group,and the autophagic bodies and lysosomes in the cytoplasm of synovial cells increased significantly in each treatment group.Compared with the blank control group,expect the low-dose group of Jinwu jiangu decoction,the expressions of LC3,Beclin-1 and VPS34 mRNA in synovial cells decreased significantly,the expression of Bcl-2 increased significantly,LC3 and Beclin-1 protein expressions were significantly decreased in each treatment group(P<0.01).The effect was more obvious in Jinwu jiangu decoction high-dose group and tripterygium glycoside group.Conclusion:Miao medicine Jinwu jiangu decoction can inhibit the expressions of autophagy-related genes LC3,Beclin-1 and VPS34 in RA synovial cells and enhance the expression level of Bcl-2.
作者
马武开
曾晨
徐晖
黄颖
唐芳
姚血明
MA Wu-kai;ZENG Chen;XU Hui;HUANG Ying;TANG Fang;YAO Xue-ming(The Second Affiliated Hospital,Guiyang College of TCM,Guiyang 550003,China)
出处
《中药材》
CAS
北大核心
2019年第4期891-896,共6页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(81360580)
贵州省高层次创新型人才培养计划——“百”层次人才项目(黔科合平台人才[2016]5650)
国家重点研发计划中医药现代化研究重点专项(2017YFC1703904)
贵州省中医药(民族医药)治疗风湿病研究平台建设(黔科合平台人才[2018]5707).
作者简介
马武开(1968-),男,博士,教授,主要从事中西医结合防治风湿病研究,Tel:0851-85556970,E-mail:walker55@163.com;通讯作者:姚血明,Tel:0851-85556970,E-mail:594305557@qq.com。
