人参皂苷Rg1对小鼠急性肝衰竭的治疗作用和机制研究  

Therapeutic efficacy and mechanism of action of ginsenoside Rgl in treating acute hepatic failure in mice

作　　者：罗欢[1] 黄文祥[1] 阳成[1] 赵金秋[1] 刘舒[1] 徐雅妹[1] 刘成伟[1] 

机构地区：[1]重庆医科大学附属第一医院感染科,400016

出　　处：《中华肝脏病杂志》2017年第3期217-222,共6页Chinese Journal of Hepatology

基　　金：基金项目：重庆市科委基础科学与前沿技术研究重点项目（CSTC2015jcyjBX0079）;重庆市教委科研项目（KJ130332）

摘　　要：目的在四氯化碳（c01,）诱导的急性肝衰竭模型中，人参皂苷Rg1（G—Rg1）对内质网应激的调节作用，以及对肝细胞凋亡的影响。方法将40只健康成年C57／BL雄性小鼠随机分为等渗盐水对照（NS）组，G-Rg1空白对照（G—Rg1）组，CCl4模型（CCl4）组，G-Rg1预防治疗（CCI＆G-Rg1）组，并建立CCl4小鼠急性肝衰竭模型。建模12h后，分别采集各组小鼠血清和肝脏组织，用试剂盒法检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总胆红素（TBil）水平；定量PCR法检测肝脏葡萄糖调节蛋白78（GRP78）、C／EBP家族同源蛋白（CHOP）的表达量；Westernb10t检测GRP78、CHOP、半胱氨酸蛋白酶12（caspase12）、caspase3的表达情况；HE染色评价肝脏组织病理学改变；免疫组织化学法分析GRP78、caspase3的表达情况；TUNEL法检测肝脏组织细胞凋亡情况。计量数据进行单因素方差分析，进一步两两比较采用LSD-t法。结果NS组、G-Rg1组、CCl4组、CCI＆G-Rg1组ALT值分别为（50．12士9．25）U／L、（40．48土6．38）U／L、（980．66±110．29）U／L和（691．30±108．06）U／L，F：365．07，P〈0．05；AST值分别为（9．69±2．78）U／L、（9．40±3．84）U／L、（319．44±89．32）U／L和（195．40土15．41）U／L，F=115．64，尸〈0．05；TBil值分别为（0．46±0．13）rag／d1（1mg／dl=17．1μmol／L）、（0．48±0．08）rag／d1、（1．56±0．12）mg／dl和（1．09±0．11）mg／dl，F=211．29，P〈0．05，CCl4＋G-Rg1组血清ALT、AST、TBil水平低于CCl4组，差异均有统计学意义。CCl4＋G-Rg1组GRP78、CHOP的mRNA相对表达量均较CCl4组下降，差异均有统计学意义（F值分别为34．4、44．1，P值均〈0．05）。Westernbolt结果显示CCl4＋G—Rg1组与cch组相比，caspase3、GRP78、caspasel2、CHOP蛋白相对表达量均有不同程度降低，差异有统计学意义（P值均〈0．05）。CClObjective To examine the regulatory effect ofginsenoside Rg1 （G-Rg1） on endoplasmic reticulum stress and its effect on hepatocellular apoptosis in carbon tetrachloride （CCl4）-induced acute liverfailure （ALF）. Methods Forty healthy, adult male C57/BL mice were randomly divided into normal saline control （NS） group, G-Rg1 blank control （G-Rg1） group, GEl4 model （CC14） group, and G-Rg1 preventive treatment （CC14＋G-Rg1） group, and an ALF mouse model was established by CC14 induction. Blood and liver specimens were collected from all mice upon sacrifice at 12 hours post-intraperitoneal injection. Serum alanine aminotransferase （ALT）, serum aspartate aminotransferase （AST） and total bilirubin （TBil） levels were determined using commercial test kits. The mRNA expression of glucose-regulated protein 78 （GRP78） and C/EBP homologous protein （CHOP） was measured using real-time PCR. The protein expression of GRP78, CHOP, caspasel2, and caspase3 were measured by Western blot. Histological changes in the liver were assessed by hematoxylin-eosin staining, and the expression of GRP78 and caspase3 was detected by immunohistochemistry. Hepatocyte apoptosis was determined using terminal transferase dUTP nick end labeling. Quantitative data were analyzed using one-way ANOVA, and subsequent pairwise comparisons were performed using the LSD-t method. Results Serum ALT, AST, and TBil levels in the CCl4＋G-Rg1 group were significantly reduced compared with those in the CCl4 group （ALT： 691.30 ± 108.06 U/L vs 980.66 ± 110.29 U/L, F = 365.07, P 〈 0.05; AST： 195.40 ± 15.41 U/L vs 319.44 ± 89.32 U/L, F= 115.64, P 〈 0.05; TBil： 1.09 ± 0.11 mg/dl vs 1.56 ± 0.12 mg/dl, F = 211.29, P 〈 0.05）. The relative mRNA expression of GRP78 and CHOP was significantly lower in the CC14 ＋ G-Rg1 group than in the CCl4 group （P 〈 0.05）. The relative protein expression of caspase3, GRP78, caspase12, and CHOP was significantly reduced to different extents in the CCl4＋G-Rg1 group

关 键 词：小鼠 细胞凋亡 肝衰竭 人参皂昔Rg1 内质网应激 

分 类 号：R575.3[医药卫生—消化系统]