苦参素抗大鼠肝纤维化作用的实验研究

Effect of Oxymatrine in CCl_4-Induced Liver Fibrosis in Rats.

目的 研究苦参素对四氯化碳诱导的大鼠肝纤维化肝组织Smad基因表达的影响。方法 90只SD大鼠分为 3组 ,正常对照组 (C组 )、模型组 (M组 )和苦参素干预组 (T组 )。以CCl4皮下注射法诱导大鼠肝纤维化 ,干预组大鼠在造模的同时给予苦参素注射液腹腔注射 ,正常对照组给予液体石蜡皮下注射和生理盐水腹腔注射 ,8周实验结束时处死动物。酶联免疫吸附法检测血清TGFβ1水平 ,HE和Masson染色观察大鼠肝组织病理学变化和胶原沉积 ,免疫组化法检测Smad3基因表达情况。结果 苦参素干预组大鼠血清TGFβ1水平下降 (32 .6 5± 8.71)vs(96 .37± 16 .5 4 )mg/L ,P <0 .0 5 ) ,肝组织组织学积分 (1.9± 0 .3vs3.6± 0 .8,P <0 .0 5 )和胶原面积 ((94 .4 1± 37.2 6 )vs(6 90 .86± 188.71) μm2 ,P <0 .0 5 )明显减少 ;Smad3蛋白表达阳性率减少 ((2 .33± 1.6 4 )vs(9.5 6± 1.34) % ,P <0 .0 5 )。结论 苦参素干预性治疗能够有效地减少大鼠肝组织胶原沉积 ,使其血清TGFβ1水平下降 ,并抑制Smad3基因表达 。

Objective To study the effect of Oxymatrine on the gene expression of Smad 3 in the prevention of cirrhosis experimentally induced in rats by chronic administration of CCl 4,and to evaluate its potential anti-fibrotic mechanisms.Methods 90 healthy male SD rats were randomly divided into three groups:the normal group(n=10),the treatment group of Salidroside(n=40)and the fibrosis group(n=40).Liver fibrosis was induced by CCl 4 subcutaneous injection(300 mL/L,3mL/kg,twice per week for 8 weeks).Treated rats received Oxymatrine celiac injection at 10mg/kg twice a week.The level of serum TGFβ1 was detected with ELISA.The deposition of collagen was observed with Masson staining.The gene expression of Smad 3 was detected with immunohistochemistry(IH)respectively.All the data were scanned and analyzed with special software.Results A reduction of collagen deposition and rearrangement of the parenchyma were noted in treated rats.The serum concentration of TGFβ1 was decreased significantly(32.65±8.71)vs(96.37±16.54)mg/L,P<0.05).The semi-quantitative histological scores(1.9±0.3vs 3.6±0.8,P<0.05)and average area of collagen((94.41±37.26)vs (690.86±188.71)μm 2,P<0.05)in treatment group were decreased.The expression of Smad 3 decreased considerably in treated animals((9.56±1.34)vs(2.33±1.64)%,P<0.05).Conclusion Oxymatrine was effective at interfering CCl 4-induced liver fibrosis in SD rats.Its potential anti-fibrotic mechanisms may be inhibiting the expression of Smad 3 thus modulating the fibrogenesis signal transduction of TGF-β,So Oxymatrine was effective in reducing the production and deposition of collagen protein in liver of experimental animal.