沙棘熊果酸通过调节线粒体-细胞色素c抑制酒精性肝病大鼠模型肝细胞凋亡的作用分析  

Ursolic acid in Hippophae rhamnoides L.inhibits hepatocyte apoptosis in rats with alcoholic liver disease by regulating mitochondria-cytochrome c

作　　者：包艳红 王强[2] 张文龙[3] 戈娜[4] 李楠 苏军 李可欣 BAO Yanhong;WANG Qiang;ZHANG Wenlong;GE Na;LI Nan;SU Jun;LI Kexin(Institute of Health,Baotou Medical College,Baotou,Inner Mongolia 014040,China;Institute of Basis and Forensic Medicine,Baotou Medical College,Baotou,Inner Mongolia 014040,China;The First Affiliated Hospital,Baotou Medical College,Baotou,Inner Mongolia 014010,China;Institute of Nutrition and Food Health,Baotou Medical College,Baotou,Inner Mongolia 014040,China;System Reform’s Office,Inner Mongolia Health Committee,Hohhot 010030,China)

机构地区：[1]包头医学院卫生健康学院,内蒙古包头014040 [2]包头医学院基础医学与法医学院,内蒙古包头014040 [3]包头医学院第一附属医院,内蒙古包头014010 [4]包头医学院营养与食品健康研究所,内蒙古包头014040 [5]内蒙古自治区综合疾病预防控制中心,呼和浩特010030

出　　处：《临床肝胆病杂志》2023年第7期1617-1626,共10页Journal of Clinical Hepatology

基　　金：国家自然科学基金(81550044,81760586);内蒙古自治区自然科学基金(2014MS0303);内蒙古自治区高等学校“青年科技英才计划”项目(NJTY-15-B11);内蒙古自治区卫生计生科研计划项目(201701084);包头医学院博士研究启动基金项目(BSJJ201630);包头医学院创新团队发展计划(BTMCTD202205);内蒙古自治区草原英才工程青年创新人才培养计划项目(Q2017089);包头市青年创新人才项目(包人社办字[2017]284号)。

摘　　要：目的 基于线粒体-细胞色素c途径探讨沙棘熊果酸对酒精性肝病大鼠肝细胞凋亡的抑制作用。方法 根据随机数字表将50只SPF级雄性Wistar大鼠进行完全随机分组,分为正常对照组、酒精模型组、沙棘熊果酸低、中和高剂量组,每组10只。正常对照组给予每日1次生理盐水灌胃8周;酒精模型组用阶梯式浓度酒精灌胃的方法持续灌胃8周;沙棘熊果酸组分别按50 mg/kg、100 mg/kg和150 mg/kg灌胃,1 h后再灌喂模型组同等剂量酒精。测定各组大鼠血清肝功能指标;HE染色观察肝组织病理情况;电镜下观察大鼠肝细胞超微结构;TUNEL法检测大鼠肝细胞凋亡情况;Western Blot法检测肝细胞线粒体和胞浆细胞色素c和活化caspase-3蛋白表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 与酒精模型组相比,沙棘熊果酸中、高剂量组大鼠血清ALT、AST和胆碱酯酶水平均下降(P值均<0.05);酒精模型组大鼠肝细胞索排列紊乱,肝细胞水肿、脂肪变性明显,而沙棘熊果酸中、高剂量组大鼠肝细胞索排列逐渐趋于正常、肝脂肪变性明显改善;肝细胞线粒体数目增加、形态明显改善;肝细胞凋亡率、胞浆细胞色素c和活化caspase-3蛋白的表达均低于酒精模型组(P值均<0.05)。结论沙棘熊果酸可改善酒精性肝病大鼠肝功能和肝组织形态,可能与抑制肝细胞线粒体细胞色素c的释放及caspase-3蛋白的活化,通过线粒体-细胞色素c途径抑制肝细胞凋亡有关。Objective To investigate the inhibitory effect of ursolic acid in Hippophae rhamnoides L.on hepatocyte apoptosis in rats with alcoholic liver disease based on the mitochondria-cytochrome c pathway.Methods A total of 50 specific pathogen-free male Wistar rats were divided into normal control group,alcohol model group,and low-,middle-,and high-dose ursolic acid groups using a random number table,with 10 rats in each group.The rats in the normal control group were given normal saline by gavage once a day for 8 weeks;the rats in the alcohol model group were given alcohol at increasing concentrations by gavage for 8 consecutive weeks;the rats in the low-,middle-,and high-dose ursolic acid groups were given ursolic acid at a dose of 50,100,and 150 mg/kg,respectively,followed by an equal volume of alcohol as the model group 1 hour later.Serum liver function parameters were measured for each group;HE staining was used to observe liver histopathology;an electron microscope was used to observe hepatocyte ultrastructure;the TUNEL method was used to measure hepatocyte apoptosis;Western Blotting was used to measure the protein expression levels of cytochrome c and activated caspase-3 in hepatocyte mitochondria and cytoplasm.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the alcohol model group,the middle-and high-dose ursolic acid groups had significant reductions in the serum level of alanine aminotransferase,aspartate aminotransferase,and cholinesterase(all P<0.05).The rats in the alcohol model group had disordered arrangement of hepatic cords with marked hepatocyte edema and fatty degeneration,while those in the middle-and high-dose ursolic acid groups had basically normal arrangement of hepatic cords and a significant improvement in hepatocyte fatty degeneration,as well as a significant increase in the number of hepatocyte mitochondria and a significant imp

关 键 词：肝疾病 酒精性 沙棘 细胞凋亡 线粒体 细胞色素C类 

分 类 号：R575.5[医药卫生—消化系统]