Hedgehog信号通路在LX-2细胞活化中的作用及慈菇消脂方的干预机制  

The role of Hedgehog signaling pathway in LX-2 cell activation and Intervention Mecha⁃nism of Cigu Xiaozhi Pills

作　　者：赵秀萍 王莉 王爱娣 马燕花[1] ZHAO Xiu-ping;WANG Li;WANG Ai-di;MA Yan-hua(First Clinical Medicine College of Gansu University of Chinese Medicine,Lanzhou Gansu 730000,China)

机构地区：[1]甘肃中医药大学第一临床医学院,甘肃兰州730000

出　　处：《时珍国医国药》2023年第4期785-790,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金地区基金(81860821);甘肃中医药大学2019创新育幼提升项目(201913-104)。

摘　　要：目的探讨Hedgehog(Hh)信号通路在LX-2细胞活化中的作用及慈菇消脂方的干预机制。方法体外培养人肝星状细胞LX-2,TGF-β1处理建立细胞活化模型。实验随机分组为空白对照组、TGF-β1诱导组即模型组(5ng/ml)、TGF-β1+含药血清组(6%)、TGF-β1+环靶明脂组(10μM),每组复设9孔,干预72h后,CCK8检测各组细胞增殖活力;Q-PCR和Western-blot检测各组细胞中Shh、Gli1、Gli2、Col-I、α-SMA mRNA和蛋白的表达水平。结果通过CCK8法确定慈菇消脂方最佳干预时间和最佳干预浓度是72h和6%含药血清。与空白对照组相比,模型组细胞活力升高(P<0.05),Shh、Gli1、Gli2、Col-I、α-SMA mRNA和蛋白表达水平升高(P<0.01)。与模型组相比,含药血清组细胞活力降低(P<0.01),Shh、Gli1、Gli2、Col-I、α-SMA mRNA和蛋白表达水平降低(P<0.05);环靶明脂组细胞活力降低(P<0.01),Shh、Gli1、Gli2、Col-I、α-SMA mRNA和蛋白表达水平降低(P<0.05)。与含药血清组相比,环靶明脂组细胞活力降低(P<0.01),Shh、Gli1、Gli2、Col-I、α-SMA mRNA和蛋白表达水平降低(P<0.05)。结论慈菇消脂方含药血清可以抑制LX-2细胞的增殖、活化,从而抗肝纤维化,其机制可能通过调控Hh信号通路相关蛋白表达。Objective To investigate the role of Hedgehog signaling pathway in LX-2 cell activation and the intervention mecha⁃nism of Cigu Xiaozhi Prescription.Methods Human hepatic stellate cells LX-2 were cultured in vitro and TGF-β1 treatment was used to establish a cell activation model.The experiments were randomly grouped into blank control group,TGF-β1 induc⁃tion group was model group(5ng/ml),TGF-β1+drug-containing serum group(6%),TGF-β1+cyclopamine group(10μM),Nine additional Wells were set in each group,after 72h of intervention,CCK8 was used to detect the proliferation of cells in each group.The expression levels of Shh,Gli1,Gli2,Col-I,α-SMA mRNA and protein in each group of cells were detected by Q-PCR and Western-blot.Results The CCK8 method determined that the optimal intervention time and optimal in⁃tervention concentration were 72h and 6%drug-containing serum.Compared with the blank control group,cell viability was in⁃creased in the model group(P<0.05),and Shh,Gli1,Gli2,Col-I,α-SMA mRNA and protein expression levels were in⁃creased(P<0.01).Compared with the model group,cell viability was reduced in the drug-containing serum group,and Shh,Gli1,Gli2,Col-I,α-SMA mRNA and protein expression levels were reduced(P<0.05);cell viability was reduced in the cyclopamine group(P<0.01),and Shh,Gli1,Gli2,Col-I,α-SMA mRNA and protein expression levels decreased(P<0.05).Compared with the drug-containing serum group,the cell viability(P<0.01)and the expression levels of Shh,Gli1,Gli2,Col-I,α-SMA mRNA and protein were reduced in the cyclopamine group(P<0.05).Conclusion The drug-contai⁃ning serum of Cigu Xiaozhi Prescription can inhibit the proliferation and activation of LX-2 cells,thus anti-liver fibrosis,and the mechanism may be through the regulation of Hedgehog signaling pathway-related protein expression.

关 键 词：肝纤维化 Hh信号通路 肝星状细胞 慈菇消脂方 

分 类 号：R285.5[医药卫生—中药学]