香砂六君丸对非酒精性脂肪肝大鼠肝脏线粒体的影响

Effect of Xiangsha Liujunzi Pill on liver's mitochondrial of rats with nonalcoholic fatty liver disease

目的:研究香砂六君丸对非酒精性脂肪肝大鼠模型肝脏组织内线粒体的影响。方法:24只正常成年雄性SD大鼠随机分为正常对照组、模型组和治疗组。采用高脂饲料喂养法来制作非酒精性脂肪肝大鼠模型,给予香砂六君丸(1.35 g·kg^(-1)·d^(-1))灌胃4周治疗后取材。检测各组大鼠的体质量、肝脏湿重、肝脏HE染色结果、血液生化指标。用Real time PCR法、紫外分光光度计法对肝脏组织线粒体DNA(mt DNA)拷贝数、线粒体复合物Ⅰ、Ⅳ活性的变化进行测定。结果:与正常对照组相比,模型组大鼠肝脏湿重和质量指数(肝脏湿重/体质量)显著升高,血清总胆固醇(TC)和低密度脂蛋白胆固醇(LDL-C)水平明显升高,肝脏组织mt DNA显著降低,肝脏组织线粒体复合物Ⅰ、Ⅳ活性显著降低,差异均有统计学意义(P<0.05)。与模型组大鼠相比,治疗组大鼠肝脏湿重和质量指数显著降低,血清TC和LDL-C水平明显降低,肝脏组织mt DNA显著升高,肝脏组织线粒体复合物Ⅰ、Ⅳ活性显著升高,差异均有统计学意义(P<0.05)。肝脏HE染色结果显示,模型组大鼠肝细胞出现大量脂肪空泡,细胞核固缩;治疗组大鼠肝细胞出现脂肪空泡量显著减少,细胞核固缩情况减轻。结论:香砂六君丸可能通过调节mt DNA拷贝数与呼吸链酶复合物活性对非酒精性脂肪肝起到治疗作用。

Objective： To observe the effect of Xiangsha Liujunzi Pill on liver’s mitochondrial in rats with nonalcoholic fatty liver disease. Methods： Under specific pathogen free （SPF） conditions, 24 male Sprague-Dawley （SD） rats were randomly divided into control group, high fat and high cholesterol diet group （HFD） and Xiangsha Liujunzi Pill （1.35 g？kg-1？d-1） treatment group. After 12 weeks, all rats were executed to get liver samples. Blood paramaters were analyzed by automatic biochemical. The copy number of mitochondrial DNA （mtDNA） and the activity of mitochondrial respiratory chain complex I, IV in liver samples was analysized with RT-PCR and UV spectrophotometer respectively. Results： Compared with control group, the hepatic wet weight and mass index （hepatic wet weight/body mass） increased significantly, levels of total cholesterol （TC） and low-density lipoprotein cholesterol （LDL-C） in serum were significantly increased, and the copy number of mtDNA and the activity of complex I, IV expression were significantly decreased in HFD group. Compared with HFD group, the liver wet weight and quality index of rats were significantly reduced, levels of TC and LDL-C in serum were significantly increased, and the copy number of mtDNA and the activity of complex I, IV expression were significantly increased in Xiangsha Liujunzi Pill treatment group. The HE staining of liver showed that the livers sections of HFD group were abundant of macrovesicular fat droplets in hepatocytes and large quantities of ballooned hepatocytes in centrilobular parenchyma with infiltration of inflammatory cells and fibrosis. The livers sections of HFD group showed that the macrovesicular fat droplets decreased and the nucleus consolidation was reduced. Conclusion： Xiangsha Liujunzi Pill would has effect on nonalcoholic fatty liver disease. Xiangsha Liujunzi Pill can regulate the disordered mitochondrial respiratory chain complex in nonalcoholic fatty liver disease.