探讨青蒿琥酯和扶正化瘀方治疗血吸虫病肝纤维化对线粒体的影响  

Exploring the effects of artesunate and fuzheng huavu decoction on mitochondria in the treatment of schistosomiasis liver fibrosis

作　　者：罗军涛 钱胜南 吴凯悦 朱思成 黄依磊 叶建平 沈双[1] Luo Juntao;Qian Shengnan;Wu Kaiyue;Zhu Sicheng;Huang Yilei;Ye Jianping;Shen Shuang(Shanghai Jiao Tong University Affiliated Sixth People's Hospital,Shanghai 200233,China;Shanghai University of Medicine&Health Sciences,Shanghai 201318,China)

机构地区：[1]上海交通大学附属第六人民医院,上海200233 [2]上海健康医学院,上海201318

出　　处：《中华肝脏病杂志》2022年第1期45-51,共7页Chinese Journal of Hepatology

基　　金：浦东新区科技发展基金(PKJ2018-Y55);上海市卫生和计划生育委员会青年项目(20174Y0124)。

摘　　要：目的比较青蒿琥酯(Art)和扶正化瘀方治疗血吸虫病肝纤维化对线粒体自噬的影响。方法将80只C57BL/6雌性小鼠随机分为健康组、感染组、Art治疗组和扶正化瘀方治疗组,每组20只。感染组及治疗组小鼠感染日本血吸虫尾蚴16条。6周后,吡喹酮(300 mg/kg)2日疗法杀虫。Art治疗组采用每天100 mg/kg腹腔注射的方式进行治疗,扶正化瘀方治疗组采用每天饲料给药,每1 kg饲料含16 g扶正化瘀方,6周后,取4组小鼠新鲜肝组织。Masson染色观察、蛋白质印迹分析肝组织中参与线粒体三羧酸循环反应的琥珀酸脱氢酶亚单位A(SDHA)、苹果酸脱氢酶(MDH2)、柠檬酸合酶(CS)、酮戊二酸脱氢酶(OGDH);雷帕霉素靶蛋白1(mTORC1)通路;腺苷酸活化蛋白激酶(AMPK)和线粒体自噬通路激酶(PINK1)的相对表达水平。提取各组肝脏组织样品的线粒体检测线粒体的耗氧率。双因素方差分析比较各组间蛋白表达的差异。结果Masson染色显示,感染组小鼠肝脏纤维化面积显著高于健康组并且Art治疗组和扶正化瘀方治疗组小鼠肝脏纤维化面积均低于感染组。蛋白质印迹法分析结果显示,SDHA蛋白相对表达量感染组(0.82±0.05)与健康组(1.00±0.05)相比显著下降(t=11.23,P=0.004),并且Art治疗组(0.73±0.05)显著高于感染组(t=10.79,P=0.007),但是扶正化瘀方组(0.98±0.05)与健康组比较,差异无统计学意义(t=1.925,P=0.127);感染组p-AMPK的蛋白质相对表达量(1.15±0.05)显著高于健康组(0.98±0.07,t=12.18,P=0.003),Art治疗组(0.50±0.05)相比较于感染组p-AMPK的表达显著降低(t=11.78,P=0.003)。感染组AMPK的蛋白质相对表达量(0.80±0.05)显著低于健康组(1.00±0.05,t=10.53,P=0.005),Art治疗组(0.54±0.05)相比较于感染组AMPK的表达显著降低(t=13.98,P=0.004);感染组p-mTORC1的蛋白质相对表达量(0.93±0.08)与健康组相比差异无统计学意义(t=2.28,P=0.065),而Art治疗组的表达量(0.63±0.05)显著低于感染组(t=Objective To compare the effects of artesunate(Art)and fuzheng huayu decoction on mitochondrial autophagy in the treatment of schistosomiasis liver fibrosis.Methods Eighty C57BL/6 female mice were randomly divided into healthy control group,infection group,Art treatment group and Fuzheng Huayu Decoction treatment group,with 20 mice in each group.Mice in the infection group and treatment group were infected with 16 Schistosoma japonicum cercariae.After 6 weeks,praziquantel(300 mg/kg)was used for 2 days to kill the wonns.The Art treatment group was treated with intraperitoneal injection of 100 mg/kg/day,while the Fuzheng Huayu Decoction treatment group was fed 16g of fuzheng huayu decoction per 1kg per day.After 6 weeks,fresh liver tissues of the four groups were collected.Masson staining and Western blot were used to observe the succinate dehydrogenase subunit A(SDHA)and malate dehydrogenase(MDH2),citrate synthase(CS),ketoglutarate dehydrogenase(OGDH),and target of rapamycin 1(mTORCl)pathway involved in mitochondrial tricarboxylic acid cycle in liver tissues.The relative expression levels of adenylate activated protein kinase(AMPK)and mitochondrial autophagy pathway kinase(PINK.1)were detected.Liver tissue samples were extracted from each group to detect the mitochondrial oxygen consumption rate.Two-way ANOVA was used to compare the significance and difference between two sets of samples.Results Masson staining showed that the infection group mice had significantly higher liver fibrosis area than the healthy control group,while the Art treatment group and Fuzheng Huayu Decoction treatment group mice had lower liver fibrosis area than the infection group.Western blot analysis showed that the infection group(0.82±0.05)had significantly lower relative expression of SDHA protein than the healthy control group(1.00±0.05)(t=11.23,P=0.0035),while the Art treatment group(0.73±0.05)had significantly higher relative expression of SDHA protein than the infection group(t=10.79,P=0.0073).However,there was no significant cha

关 键 词：肝纤维化 线粒体 青蒿琥酯 扶正化瘀方 

分 类 号：R285.5[医药卫生—中药学]