藏药二十五味松石丸减方对大鼠慢性肝纤维化的防治作用研究

Effect of Tibetan medicine Ershiwuwei Songshi pills modified decoction on rats liver fibrosis

目的:探讨二十五味松石丸减方前后对大鼠慢性肝纤维化的防治作用。方法:Wistar雄性大鼠,随机分为模型组、空白对照组、阳性对照组(复方鳖甲软肝片630mg/kg)、二十五味松石丸105mg/kg组、原方减朱砂103mg/kg组、原方减松石98mg/kg组、原方减朱砂和松石97mg/kg组。除空白对照组以外,其余各组大鼠皮下注射40%CCL_4橄榄油溶液连续造模8周,造模期间模型组和空白对照组灌胃等量的生理盐水,其余各组灌胃给予相应药物,8周末处死大鼠。测定大鼠血清指标谷草转氨酶(AST)、白蛋白(AL)、谷丙转氨酶(ALT)含量,采用ELISA法测定大鼠透明质酸(HA)、羟脯氨酸(HYP)、层粘连蛋白(LN)、转化生长因子-β1(TGF-β1),测定超氧化歧化酶(SOD)的水平。进行常规HE染色观察各组大鼠肝组织病理学变化,采用Masson染色观察各组大鼠肝纤维化的程度。结果:与空白对照组比较,模型组大鼠血清ALT、AST、TGF-β1、HA、LN和HYP水平显著升高,AL含量明显降低。与模型组比较,二十五味松石丸105mg/kg组、原方减朱砂和松石97mg/kg组大鼠血清ALT、TGF-β1、HA、LN和HYP水平明显降低,AL含量明显升高,肝纤维化面积明显减少。结论:二十五味松石丸减方前后均对CCL_4所致大鼠慢性肝纤维化具有一定的防治作用,以二十五味松石丸组的防治作用最佳,原方减朱砂和松石组次之,原方减朱砂组、原方减松石组的防治作用均最差。

Objective: In this study,we investigated the prevention and treatment effects of Ershiwuwei Songshi pills modified decoction on rats liver fibrosis. Methods: Wistar rats were randomly divided into 7 groups: Model group,Normal control group,Positive control group( 630 mg/kg Fufangbiejiaruangan pills),105 mg/kg Ershiwuwei Songshi pills group,103 mg/kg Ershiwuwei Songshi pills without cinnabar group,98 mg/kg Ershiwuwei Songshi pills without Songshi group and 97 mg/kg Ershiwuwei Songshi pills without Songshi and cinnabar group. Except for the normal control group,rats in other groups were subcutaneously injected with olive oil solution containing 40% CCL4 for 8 weeks. During the course,rats in the model group and the normal control group were orally administered with 1. 5 mL normal saline,while rats in other groups were administered with corresponding drugs. After 8 weeks,all rats were sacrificed. The levels of ALT,AST,and AL in serum were determined by biochemical analysis,while levels of transforming growth factor-β1( TGF-β1),hyaluronic acid( HA),laminin( LN),hydroxyproline( HYP),and superoxide dismutase( SOD) were determined by enzyme-linked immuno sorbent assay( ELISA). The liver tissue morphology was observed by HE staining. Hepatic fibrosis was evaluated by Masson trichrome staining. Results: Compared with Normal control group,the levels of ALT,AST,TGF-β1,HA,LN and HYP in Model group were significantly increased. Compared with Model group,the levels of AL in Ershiwuwei Songshi pills group and Ershiwuwei Songshi pills without Songshi and cinnabar group were significantly increased,the levels of TGF-β1,HA,LN and HYP in Ershiwuwei Songshi pills group and Ershiwuwei Songshi pills without Songshi and cinnabar group were significantly decreased,and the area of hepatic fibrosis was significantly reduced. Conclusion: The results indicate that they have certain prevention and treatment effects on rats liver fibrosis before and after Ershiwuwei Songshi pills is modified. The intensities of effects show as following: Ershiwuwei Songshi pills > Ershiwuwei Songshi pills without Songshi and cinnabar > Ershiwuwei Songshi pills without cinnabar and Ershiwuwei Songshi pills without Songshi.