青蒿琥酯抗大鼠免疫性肝纤维化的作用及机制研究

Effects and mechanisms of artesunate on immunological hepatic fibrosis in experimental rats

目的研究青蒿琥酯抗实验性肝纤维化的作用及其可能机制。方法制备牛血清白蛋白免疫性大鼠肝纤维化模型。动物随机分为6组:正常对照组、模型对照组、Art组(5、15、45 mg.kg-1)、阳性对照Col组(0.1 mg.kg-1)。Masson染色观察肝组织病理变化,Jamall法测定肝组织中羟脯氨酸(Hyp)的含量,RT-PCR技术检测肝组织转化生长因子β1(TGF-β1),α-肌动蛋白(α-SMA)mRNA的表达。体外培养大鼠肝星状细胞株(HSC-T6),用TGF-β1(5μg.L-1)刺激,与Art(终浓度6.25、25、50 mg.L-1)共培养后,检测HSC-T6中Ⅰ型前胶原(procollagenⅠ)mRNA及Ⅰ型胶原(collagenⅠ)蛋白的表达。结果与模型组相比,Art各组能减轻肝组织纤维增生的程度,降低肝组织Hyp含量,使肝组织α-SMA、TGF-β1 mRNA和HSC-T6中procollagenⅠmRNA和collagenⅠ蛋白的表达水平下降。结论 Art有明显的抗肝纤维化作用,其机制可能与抑制HSC活化,降低TGF-β1 mR-NA及collagenⅠ蛋白的表达有关。

Aim To investigate the effects and mechanisms of artesunate on immunological hepatic fibrosis in experimental rats.Methods The model of immunological hepatic fibrosis induced by bovine serum albumin was prepared.The rats were randomly divided into 6 groups： the normal group,the model group,the positive control group treated by colchicine（0.1 mg·kg-1）,and the three artesunate groups（5,15,45 mg·kg-1）.The pathological morphology of liver was examined by masson trichrome stain and the content of Hyp were analyzed via jamall′s method;The expressions of TGF-β1 mRNA and α-SMA mRNA in liver were detected by RT-PCR.Hepatic stellate cells（HSC-T6） were incubated in vitro,stimulated by TGF-β1（5 μg·L-1） and treated with different concentrations of Art（6.25,25,50 mg·L-1） for 24 h.The expressions of procollagen Ⅰ mRNA and collagen Ⅰ protein in HSC-T6 cells were separately determined by RT-PCR and Western blot.Results The degree of hepatic fibrosis and the content of Hyp in artesunate group were reduced in comparison with that of the model group.The expressions of TGF-β1 mRNA and α-SMA mRNA were decreased in artesunate groups.The expressions of procollagen Ⅰ mRNA and collagen Ⅰ protein were down-regulated in artesunate groups.Conclusion Artesunate significantly inhibited the progress of hepatic fibrosis in experimental rats,probably by inhibiting the activation of HSC and down-regulating the expression of TGF-β1 mRNA and collagenⅠprotein.