穿心莲内酯和吡非尼酮联合给药对TGF-β1诱导的肝星状细胞活化的影响及作用机制  

Study on the Effect and Mechanism of Andrographolide Combined with Pirfenidone on the Activation of Hepatic Stellate Cells Induced by TGF-β1

作　　者：许光[1] 马体栋[1] 赵凡[1] 李碧香[1] 周崇高[1] XU Guang;MA Ti-dong;ZHAO Fan(Department of Neonatal Surgery,Hunan Children’s Hospital,Changsha Hunan 410007)

机构地区：[1]湖南省儿童医院新生儿外科,湖南长沙410007

出　　处：《医学临床研究》2022年第2期168-172,共5页Journal of Clinical Research

基　　金：湖南省中医药科研计划项目(2020048);湖南省出生缺陷协同防治科技重大专项(2019SK1010)。

摘　　要：【目的】探讨穿心莲内酯(AG)联合吡非尼酮(PFD)对转化生长因子-β1(TGF-β1)诱导的人肝星状细胞LX-2活化的影响及作用机制。【方法】以LX-2作为研究对象,常规体外培养后分为Control组、TGF-β1组、AG组、PFD组、AG+PFD组。采用MTT法测定各组细胞活力,采用qRT-PCR检测各组纤维标志物和炎性标志物mRNA表达水平,采用Western blot检测各组细胞中TGF-β/Smad信号通路相关蛋白表达水平。【结果】相对于TGF-β1组,AG组、PFD组和AG+PFD组中纤维标志物mRNA[α-平滑肌肌动蛋白(α-SMA)mRNA、Ⅰ型胶原蛋白(Collagen-Ⅰ)mRNA、结缔组织生长因子(CTGF)mRNA],炎性标志物mRNA[白细胞介素6(IL-6)mRNA、白细胞介素1β(IL-1β)mRNA、肿瘤坏死因子(TNF-α)mRNA]和TGF-β/Smad信号通路相关蛋白(p-Smad2、p-Smad3)显著下调(均P<0.05);相对于AG组、PFD组,AG+PFD组中Collagen-ⅠmRNA、CTGF mRNA、p-Smad2和p-Smad3表达水平显著下调(均P<0.05);相对于PFD组,AG+PFD组中α-SMA mRNA、IL-6 mRNA表达水平显著下调(均P<0.05)。【结论】与单一疗法相比,AG联合PFD对TGF-β1诱导的LX-2活化发挥了更好的抗肝纤维化作用,其作用机制可能与其调控TGF-β/Smad信号通路有关。【Objective】To investigate the effect of andrographolide(AG)combined with pirfenidone(PFD)on hepatic stellate cell activation induced by transforming growth factor-β1(TGF-β1)and its mechanism.【Methods】Hepatic stellate cells(LX-2)were used as the research object.After routine in vitro culture,they were divided into control group,TGF-β1 Group,AG group,PFD group and AG+PFD group.MTT assay was used to determine the cell viability of five groups,RT-PCR was used to detect the mRNA expression levels of fiber markers and inflammatory markers,and Western blot was used to detect the related protein expression level of TGF-β/Smad signaling pathway cells in each group.【Results】Compared with TGF-β1 group,fiber markers(α-SMA mRNA,Collagen-ⅠmRNA and CTGF mRNA)inflammatory marker(IL-1βmRNA,IL-6 mRNA and TNF-αmRNA)and TGF-β/Smad signaling pathway related proteins(p-Smad2,p-smad3)of the AG group,the PFD group and the AG+PFD group were significantly down regulated(mean P<0.05);Compared with the PFD group,the expression levels ofα-SMA mRNA and IL-6 mRNA in the AG+PFD group were significantly down regulated(mean P<0.05).【Conclusion】Compared with monotherapy,AG combined with PFD therapy has significant effect on the activation of hepatic stellate cells induced by TGF-β1,and plays a better role in anti hepatic fibrosis,and its mechanism may be related to the regulation of TGF-β/Smad signaling pathway.

关 键 词：肝/细胞学 星形细胞 转化生长因子β1 吡啶酮类 穿心莲内酯 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学]