地顶孢霉菌丝体对免疫性肝纤维化大鼠的保护作用及其可能机制的研究

摘要

To investigate the protective effects of Acremoniumterricola milleretal mycelium( AMM ) on immunological hepatic fibrosis rats induced by porcine serum and its mechanism. Methods Adult healthy of SD male rats were randomly divided into six groups: normal control group, liver fibrosis model group, colchicines group ( 0. 1 mg/kg ) and AMM( 700,350,175 mg/kg ) treated groups. Establishing a rat model of liver fibrosis by giving an intraperitoneal injection of porcine serum ( 0. 5 ml,twice a week )for 18 weeks. Except for normal control group with same volume of normal saline. The drug was given by intragastrically in AMM group and colchicines group respectively , and rats in the normol control group and model group were given the same volume of distilled water. VG staining were used to examine the histopathological change. The activity of ALT, AST in sera and the concent of Hyp in hepatic tissue were measured by commercial kits. The content of TGF-β1 was measured by ELISA. The expression of TGF-β1 protein in the liver tissues was assayed by immunohistochemistry and Western blot respectively. Results Compared with hepatic fibrosis model group, VG staining showed that AMM( 700,350 mg/kg )significantly alleviated collagen area of liver constitution ( P < 0. 05 ,P < 0. 01 ). Moreover, AMM treatment significantly reduced content of Hyp,TGF-β1( P <0. 05 ,P <0. 01 ). The results of immunohistochemistry and Western blot showed that AMM( 700,350 mg/kg )significantly decreased TGF-β1 protein expression levels of liver tissues in immune hepatic fibrosis rats( P < 0. 05 ,P < 0. 01 ). Conclusion AMM has significantly against immune-mediated liver fibrosis in rats by the way of reducing TGF-β1 synthesis and secretion, which may be one of the anti-hepatic fibrosis mechanisms of AMM.%目的 探讨地顶孢霉菌丝体(AMM)对猪血清诱导的免疫性肝纤维化大鼠的保护作用及机制.方法 健康成年SD大鼠90只,随机分为空白对照组、模型组、秋水仙碱组(Col 0.1 mg/kg)、AMM组(700、350、175 mg/kg).除空白对照组腹腔注射等量的生理盐水外,其余各组分别腹腔注射猪血清0.5 ml/只,每周2次,连续18周,建立肝纤维化模型.并于造模之日起,AMM组和Col组灌胃给药,空白对照组和模型组分别给予等量蒸馏水.VG染色观察AMM对肝脏病理形态学影响;测定血清中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)的活性及肝脏匀浆中羟脯氨酸(Hyp)的含量;ELISA法测定血清中转化生长因子β1(TGF-β1)的含量;免疫组化和Western blot法检测大鼠肝脏组织TGF-β1蛋白表达水平.结果 VG染色显示,与模型组比较,AMM(700、350 mg/kg)组肝组织胶原面积明显减少,TGF-β1和Hyp含量均显著降低(P<0.05,P<0.01);免疫组化和Western blot结果 显示,AMM(700、350 mg/kg)明显下调TGF-β1表达水平(P<0.05,P<0.01).结论 AMM能有效抗免疫性肝纤维化,其机制可能与下调肝纤维化大鼠中升高的TGF-β1的表达和分泌有关.