基于细胞自噬研究双氢青蒿素对肝星状细胞活化的影响

Effects of Dihydroartemisinin(DHA)on Activation of Hepatic Stellate Cells(HSCs)Based on Autophagy

目的:基于细胞自噬探讨双氢青蒿素(DHA)对肝星状细胞(HSC)活化的影响。方法:体外培养HSC-T6细胞,分为正常细胞对照组、DHA组、3-MA组、DHA+3-MA组共4组。用免疫组化法检测HSC-T6细胞α-平滑肌肌动蛋白(α-SMA)的表达。定量聚合酶链反应(PCR)检测HSC-T6细胞Ⅰ型胶原和转化生长因子-β1(TGF-β1)的m RNA表达。结果:正常培养的HSC大量表达α-SMA,经DHA(6.25μg/m L)或自噬抑制剂3-MA(5.0 mmo L/L)干预后,α-SMA表达明显减少,而经DHA和3-MA联合处理,α-SMA表达进一步减少。DHA和3-MA均能下调HSC表达Ⅰ型胶原和TGF-β1m RNA,与对照组比较,差异具有统计学意义(P<0.05)。而DHA和3-MA联合干预,Ⅰ型胶原和TGF-β1 m RNA表达均进一步下降,与DHA组或3-MA组比较,差异具有统计学意义(P<0.05)。结论:DHA可抑制HSC活化和增殖,从而在体外发挥抗肝纤维化作用,而细胞自噬能促进这一进程。

Objective To investigate the effects of dihydroartemisinin（DHA）on activation of hepatic stellate cells（HSCs）based on autophagy. Methods The cultured HSC-T6 cells were used for experimental cells,and were divided into four groups ：normal control group,DHA group,3—methyladenine（3-MA）group and DHA ＋ 3-MA group. Immunohistochemical assay was used for the detection of α-smooth muscle actin（α-SMA） expression. Quantitative polymerase chain reaction（PCR） was applied for the detection of collagen type Ⅰ and transforming growth factor-β1（TGF-β1） m RNA expressions of HSC-T6 cells.Results HSCs of normal control group express α-SMA in great amount. By DHA（6.25μmol/L）or 3-MA（5.0 mmol/L）therapy,α-SMA expression of HSCs was decreased significantly. By combined therapy of DHA and 3-MA,α-SMA expression of HSCs was decreased further.Compared with the normal control group,the collagen type I and TGF-β1 m RNA expression in DHA group or 3-MA group were downregulated（all P〈 0.05）. Compared with the DHA group or 3-MA group, the collagen type I and TGF-β1 m RNA expression in DHA ＋ 3-MA group was significantly decreased further（all P 〈0.05）. Conclusions The in-vitro anti-liverfibrosis mechanism of DHA may be related with the inhibition of activation and proliferation of hepatic stellate cells,while autophagy may accelerate the course.