芍药苷抑制TGF-β1诱导的肝星状细胞活化及自噬

Paeoniflorin suppresses TGF-β1-induced hepatic stellate cell activation and autophagy

目的:研究芍药苷对转化生长因子-β1(TGF-β1)诱导的肝星状细胞株LX-2活化及自噬的影响,初步探讨芍药苷抗肝纤维化的作用机制。方法:采用不同浓度芍药苷分别作用于LX-2及TGF-β1诱导的LX-2细胞,用MTT法检测芍药苷对细胞活力、细胞增殖的影响,选出最合适的芍药苷干预浓度;采用流式细胞学实验及MTT法检测芍药苷对细胞凋亡及增殖的影响;蛋白免疫印迹法检测芍药苷对细胞α-平滑肌肌动蛋白(α-SMA)、LC3-Ⅰ/Ⅱ、Beclin 1、Atg5、趋化因子CXC配体2(CXCL12)、趋化因子受体4(CXCR4)、β-catenin蛋白水平的变化;ELISA法检测芍药苷对细胞炎症因子TNF-α、CXCL12、IL-1β、IL-6的影响。结果:芍药苷能抑制TGF-β1诱导的LX-2细胞的活性及生存率(P<0.05)。芍药苷能促进LX-2细胞的凋亡,抑制LX-2细胞的增殖;与对照组相比,TGF-β1诱导的LX-2细胞α-SMA、LC3-Ⅰ/Ⅱ、Beclin 1、Atg5、CXCL12、CXCR4、β-catenin蛋白水平及TNF-α、CXCL12、IL-1β、IL-6水平明显增加(P<0.05),而芍药苷预处理能逆转上述效应(P<0.05)。TGF-β1能促进LX-2细胞炎症因子TNF-α、CXCL12、IL-1β、IL-6的表达(P<0.05),而芍药苷预处理能抑制上述炎症因子的表达水平(P<0.05)。结论:芍药苷能抑制TGF-β1诱导的肝星状细胞活化及自噬水平。

Objective:To study the effect of paeoniflorin(PAE)on transforming growth factor-β1(TGF-β1)-induced activation and autophagy of LX-2 in hepatic stellate cells and to explore the mechanism of the anti-liver fibrosis effect of paeoniflorin.Methods:Different concentrations of paeoniflorin were used to act on LX-2 and TGF-β1-induced LX-2 cells,and the effects of paeoniflorin on cell viability and cell proliferation were detected by MTT and to select the most suitable intervention concentration of paeoniflorin;flow cytometry and MTT were used to detect the effects of paeoniflorin on cell apoptosis and and cell proliferation.The protein levels ofα-smooth muscle actin(α-SMA),LC3-I/II,Beclin 1,Atg5,CXCL12,CXCR4 andβ-catenin were detected by Western blot.The cellular inflammatory factors levels of TNF-α,CXCL12,IL-1β,IL-6 were determined by ELISA.Results:PAE inhibited the activity and survival of LX-2 cells(P<0.05).Furthermore,Paeoniflorin significantly promoted the apoptosis and inhibited proliferation of LX-2 cells.Moreover,compared with the control group,α-SMA,LC3-Ⅰ/Ⅱ,Beclin 1,Atg5,CXCL12,CXCR4,β-catenin protein levels of TGF-β1-induced LX-2 cells and TNF-α,CXCL12,IL-1β,IL-6 levels were significantly increased(P<0.05),However,these effects were reversed peonidin pretreatment(P<0.05).TGF-β1 treatment elevated the expression of inflammatory factors TNF-α,CXCL12,IL-1β,IL-6 in LX-2 cells(P<0.05),while paeoniflorin treatment inhibited the expression levels of these inflammatory factors(P<0.05).Conclusion:Paeoniflorin suppresses TGF-β1-induced hepatic stellate cell activation and autophagy.