牛大力水提取物抗大鼠肝纤维化作用的研究

Study on the anti-hepatic fibrosis effect of aqueous extracts from Millettia speciosa

目的:探究壮药牛大力抗大鼠肝纤维化作用及其作用机制。方法:开展体外细胞实验,利用CCK8法观测牛大力水提物对肝星状细胞(HSC)增殖的抑制作用,运用流式细胞仪检测HSC的凋亡情况。体内动物实验则采用复合因素制备肝纤维化大鼠模型,灌胃给予牛大力水提物3周后,测定血清中Ⅳ型胶原(Ⅳ-C)、层黏蛋白(LN)、谷胱甘肽(GSH)以及转化生长因子-β_(1)(TGF-β_(1))、肿瘤坏死因子-α(TNT-α)的蛋白表达,并观察肝组织的病理变化。结果:在体外细胞实验中,牛大力水提物能抑制HSC的增殖,诱导HSC发生凋亡,且随着水提物浓度增加作用增强。而在动物体内实验中,牛大力水提物组的LN水平显著降低,GSH的水平显著增加(P<0.01),但只有高、中剂量组的Ⅳ-C水平出现显著性降低(P<0.01)。另外,高剂量组的TGF-_(1)表达减少(P<0.05)。通过Masson染色和HE染色发现,牛大力水提物能改善肝纤维化大鼠肝组织的病变程度。结论:牛大力水提物具有抗肝纤维化作用。

OBJECTIVE To study and explore the effect and the mechanism of aqueous extracts from Millettia speciosa antirat liver fibrosis.METHODS In vitro cell experiments were carried out to observe the inhibitory effect of bovine Dali aqueous extract on the proliferation of hepatic stellate cells(HSCs)by CCK8 method,and the apoptosis of HSCs was detected by flow cytometry.A rat model of liver fibrosis was prepared by using multiple factors.After gavage of aqueous extracts from Millettia speciosa the serum levels ofⅣ-C,LN,GSH and the protein expression of TGF-β_(1)and TNT-αwere determined.Masson staining and HE staining were performed on rat liver tissues to observe the pathological changes of liver tissues.RESULTS In vitro cell experiment,the aqueous extracts from Millettia speciosacould inhibit the proliferation of HSCs and cause HSC to undergo apoptosis,and the effect increased with the increase of the concentration of the aqueous extracts.In animal experiments,the level of LN in the High-concentration aqueous extracts from Millettia speciosa was significantly reduced,and the level of GSH was significantly increased(P<0.01).As for the impact ofⅣ-C levels,only the high and medium dose groups showed a significant decrease(P<0.01).In addition,the expression of TGF-β_(1)in the high-dose group was reduced(P<0.05).Through Masson staining and HE staining,it was found that the water extract of Millettia speciosa can improve the degree of pathological changes of liver tissue in rats with liver fibrosis.CONCLUSION Taqueous extracts from Millettia speciosa has anti-liver fibrosis effect.