三七皂苷Rg1对肝纤维化大鼠线粒体质子跨膜转运的作用机制

Action mechanism of proton translocation across mitochondrial membrane of notoginsenoside Rg1 on hepatic fibrosis rats

目的探讨在三七皂苷(panax notoginseng saponins,Pn S)Rg1干预下,肝纤维化大鼠线粒体质子跨膜转运的变化和肝线粒体膜的流动性的变化,为开发三七单体Rg1在临床抗纤维化的应用提供详尽的试验依据和理论基础。方法 72只Wistar大鼠随机分为对照组、四氯化碳(carbon tetrachloride,CCl4)肝纤维化大鼠模型组、Pn S Rg1组各24只。除对照组外,其余2组用5%CCl4橄榄油按5 m L/kg灌胃制作肝纤维化大鼠模型。Pn S Rg1组在每次CCl4灌胃同时腹腔注射Pn S Rg1(5 mg/kg)用稳态荧光探针标记技术动态观察肝纤维化大鼠线粒体质子跨膜转运的变化,用荧光偏振法测定肝线粒体膜的流动性和膜的微黏度的改变。结果 (1)与对照组相比,肝纤维化模型组大鼠用单因素多组间方差分析,发现模型组大鼠质子跨膜转运中,肝线粒体质子跨膜转运能力显著下降(P<0.01)。Pn S Rg1组与对照组相比质子跨膜转运的变化没有显著性意义(P>0.05),与模型组相比,有显著性差异(P<0.01)。(2)肝纤维化模型组大鼠用单因素多组间方差分析,与对照组相比,证实模型组大鼠线粒体膜的流动性显著下降(P<0.01),增加膜的微黏度(P<0.01);而Rg1组与模型组相比增加线粒体膜的流动性(P<0.01),降低膜的微黏度(P<0.01)。结论肝纤维化大鼠肝线粒体质子跨膜转运能力下降和线粒体膜的流动性显著下降是导致肝纤维化的重要原因之一。Pn S Rg1通过增加肝线粒体质子跨膜转运能力和线粒体膜的流动性而防治肝纤维化的发生发展的。

Objective To investigate the changes in proton translocation across mitochondrial membrane and hepatic mitochondria membrane fluidity of rats with hepatic fibrosis under the intervention of notoginsenoside Rg1, in order to provide detailed experimental and theoretical basis for the clinical applica-tion of notoginsenoside Rg1. Methods 72 Wistar rats were randomly divided into control group, carbon tetrachloride （ CCl4 ） group, model group and notoginsenoside Rg1 group, and with 24 rats in each group. All the rats received gavage administration with 5% CCl4 solution （5 mL/kg） in addition to the control group, and rats in notoginsenoside Rg1 group received intraperitoneal injection with notoginsenoside Rg1 （5 mg/kg） based on the gavage administration. The change of proton translocation across mitochondrial mem-brane of rats with hepatic fibrosis was observed dynamically by using steady-state fluorescence probe tech-nique. The change of hepatic mitochondria membrane fluidity and the viscosity of membrane were observed by using fluorescence polarization methods. Results （1）Compared with control group, the ability of pro-ton-translocation across mitochondrial membrane in hepatic was declined significantly of rats in model group （ P〈0. 01 ） . There was no significant difference in ability of proton-translocation across mitochondrialnbsp;membrane in hepatic of rats between control and notoginsenoside Rg1 groups （P〉0. 05）, while the differ-ence was significant between model and notoginsenoside Rg1 groups （ P〈0. 01 ） . （ 2 ） Compared with con-trol group, the mitochondrial membrane fluidity of rats in model group was declined significantly （ P 〈0. 01）, and the membrane viscosity was increased significantly （P〈0. 01）. Compared with model group, the mitochondrial membrane fluidity of rats in notoginsenoside Rg1 group was increased （ P〈0. 1 ） , while the membrane viscosity was declined significantly （ P〈0. 01 ） . Conclusion One of the important causes of hepatic fibrosis rats was the decline in ability of proton-translocation across mitochondrial membrane and membrane fluidity in hepatic. Notoginsenoside Rg1 could prevent the occurrence and development of hepat-ic fibrosis through increasing the ability of proton-translocation across mitochondrial membrane and mem-brane fluidity of hepatic.