三七皂甙Rg1、Rb1抗大鼠肝纤维化的作用及机制研究

Study on the effect and action mechanism of panax notoginseng saponins-Rg1,Rb1 on hepatic fibrosis in rats

目的探讨三七皂甙Rg1、Rb1对肝纤维化大鼠线粒体的保护作用及机制。方法将大鼠随机分为4组,分别为对照组、模型组、Rg1、Rb1保护组,用5%CCl4橄榄油胃饲复制大鼠肝纤维化模型,用透射电镜和光镜观察肝细胞形态学变化。用放射免疫法检测Ⅲ型前胶原(procollagen typeⅢ,PCⅢ)的水平。用[3H]油酸标记的生物膜法测定肝线粒体分泌型磷脂酶A2(secretory phospholipase A2,sPLA2)活性,用分光光度法测定肝线粒体MDA含量,SOD活性,Ca2+-ATP酶,总ATP酶活性,用荧光偏振法测定肝线粒体膜的流动性,紫外分光光度法测定呼吸链复合体Ⅰ,Ⅱ,Ⅲ,Ⅳ的活性。结果组织学用电镜检测可见模型组细胞有粗大束状的胶原纤维沉淀。HE染色和Masson胶原染色切片统计分析得出Rg1、Rb1组的纤维化程度与对照组比较有统计学意义(P<0.01)。血清学检查显示Rg1、Rb1组显著抑制血清中ALT、PCⅢ的升高,提高线粒体SOD活性,降低MDA含量,抑制sPLA2的升高,提高线粒体膜Ca2+-ATP酶活性,总ATP酶活性(P<0.01);生物物理学证实Rg1、Rb1组增加线粒体膜的流动性,降低膜的微粘度,显著增加呼吸链复合体Ⅲ,Ⅳ的活性(P<0.01)。结论三七皂甙Rg1、Rb1通过对肝纤维化大鼠线粒体显著的保护机理而起到抗肝纤维化的作用。

Objective To investigate the protective effect panax notoginseng saponins -Rg1, Rb1 on mitochondria of rats with hepatic fibrosis .Methods The rats were randomly divided into four groups , control group,model group,Rg1 group and Rb1group.The rat models with hepatic fibrosis were established by giving 5%CCI4 solution via gavage , then hepatic morphological changes were observed by transmission electron microscope and light microscope;procollagen type Ⅲ （ PCⅢ） levels were detected by radio-immuno assay;secretory phospholipase A2 （sPLA2） activity was tested by labeled E.coli membrane with [3H] oleic acid.The changes of mitochondrial membrane fluidity were detected by fluorescent polarization;MDA content,activities of SOD,Ca2＋-ATPase and total ATPase were determined by spectrophotography;activities of respiratory chain complex Ⅰ,Ⅱ,Ⅲ,Ⅳ were detected by ultraviolet spectrophotometry .Results Transmission electron microscope showed that collagen fibrils and rough bunch of collagen fibrils precipitation were found in control group .The results of HE staining and Masson collagen staining slices demonstrated that there were significant differences in fibration degrees between Rg1,Rb1 groups and control group （ P 〈0.01）.Serological testing showed that Rg 1 and Rb1 remarkably reduced the levels of ALT ,PCⅢin serum,enhanced SOD activity and decreased MDA levels .Rg 1 and Rb1 also significantly increased the activities of respiratory chain complex Ⅰ,Ⅲ,Ⅳ and total ATPase （ P 〈0.01）. Conclusion Panax notoginseng saponins-Rg1,Rb1 has inhibitory effect on hepatic fibrosis through protecting the mitochondria of rats with liver fibrosis .