草乌甲素对乙醛诱导的LX-2细胞ECM分泌及相关蛋白表达的影响  

Effects of bulleyaconitine A on extracellular matrix secretion and expression of related proteins in acetaldehyde-induced LX-2 cells

作　　者：袁亚平 孙启山 周小倩 姚润 湛孝东[1] 赵金红[1] 姜玉新 唐小牛[1] YUAN Yaping;SUN Qishan;ZHOU Xiaoqian;YAO Run;ZHAN Xiaodong;ZHAO Jinhong;JIANG Yuxin;TANG Xiaoniu(Department of Medical Parasitology,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]皖南医学院医学寄生虫学教研室,安徽芜湖241002 [2]嘉兴学院医学院病原生物学与免疫学教研室,浙江嘉兴314001

出　　处：《皖南医学院学报》2022年第5期416-419,共4页Journal of Wannan Medical College

基　　金：国家自然科学基金项目(81671586);安徽省高校自然科学研究重点项目(KJ2018A0263,KJ2019A0403)。

摘　　要：目的:探讨草乌甲素(BLA)对乙醛诱导的人LX-2肝星状细胞(LX-2细胞)外基质及其相关蛋白的影响。方法:将细胞分为空白对照组(NC组)、乙醛组(模型组)和BLA+乙醛组(BLA组),乙醛浓度为400μmol/L,BLA浓度为18.75μg/mL。对NC组和BLA组LX-2细胞进行形态学评价和DAPI染色,ELISA检测三组Ⅰ型胶原蛋白(Col-Ⅰ)和Ⅲ型胶原蛋白(Col-Ⅲ)的浓度,WB检测三组α-SMA、TGF-β1、MMP-1、TIMP-1的蛋白含量。结果:乙醛组Col-Ⅰ和Col-Ⅲ的含量高于对照组(P<0.05),而BLA组Col-Ⅰ、Col-Ⅲ含量低于乙醛组(P<0.05)。乙醛组TGF-β1、α-SMA和TIMP-1蛋白含量均高于对照组(P<0.05),而MMP-1蛋白含量低于对照组(P<0.05);BLA组TGF-β1、α-SMA和TIMP-1蛋白含量均低于乙醛组(P<0.05),而MMP-1蛋白含量高于乙醛组(P<0.05)。结论:BLA可能通过抑制TGF-β1信号通路来抑制LX-2细胞的激活和增殖,并通过降低MMP-1/TIMP-1比值来降低Col-Ⅰ和Col-Ⅲ的含量。Objective:To investigate the effect of bulleyaconitine A(BLA) on the production of extracellular matrix secretion(ECM) and related proteins by acetaldehyde-activated LX-2 human hepatic stellate cells(LX-2 cells).Methods:LX-2 cells were divided into blank control group(NC group),acetaldehyde group(Model group) and BLA + acetaldehyde group(BLA group).Dose of acetaldehyde and BLA used in the experiment was 400 μmol/L and 18.75 μg/mL,respectively.LX-2 cells in NC group and BLA group were subjected to morphological evaluation and DAPI staining.The concentrations of collagen type Ⅰ(Col-Ⅰ) and collagen type Ⅲ(Col-Ⅲ) in the three groups were detected by ELISA,and the protein contents of α-SMA,TGF-β1,MMP-1 and TIMP-1 in the three groups were detected by WB.Results:Collagen type I(Col-Ⅰ) and collagen type Ⅲ(Col-Ⅲ) content were significantly higher in model group than in NC group(P<0.05),whereas markedly lower in BLA group than in model group(P<0.05).The expression levels of TGF-β1,α-SMA and TIMP-1 in model group were significantly higher than those in the NC group(P<0.05),yet MMP-1 expression was lower than that in NC group(P<0.05).TGF-β1,α-SMA and TIMP-1 were highly expressed in BLA group compared to model group(P<0.05),whereas higher MMP-1 expression was found in model group(P<0.05).Conclusion:BLA may suppress the activation and proliferation of LX-2 cells via inhibiting TGF-β1 signalling pathway, and down-regulate Col-Ⅰ and Ⅲ content through reducing MMP-1/TIMP-1 ratio.

关 键 词：草乌甲素 LX-2细胞 肝纤维化 

分 类 号：R285.5[医药卫生—中药学]