特异性敲除CD147基因抑制小鼠非酒精性脂肪性肝炎的机制研究  

Mechanism of specific knockout of CD147 to inhibit non-alcoholic steatohepatitis

作　　者：沈晓雯[1] 戴世荣[1] 黄琳玲 SHEN Xiao-wen;DAI Shi-rong;HUANG Liri-ling(Department of Laboratory,the Second People's Hospital of Nantong in Jiangsu ( Nantong Jiangsu,226001 ) China)

机构地区：[1]江苏省南通市第二人民医院检验科,江苏南通226001

出　　处：《中西医结合肝病杂志》2019年第1期50-53,I0008,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：南通市科技计划项目(No.YY215028)

摘　　要：目的:研究特异性敲除小鼠CD147基因抑制其非酒精性脂肪性肝炎(NASH)的可能作用机制。方法:选择8周龄小鼠24只,其中12只敲除CD147基因,另12只不敲除。将敲除CD147基因的12只小鼠分为两组,每组6只。其中1组小鼠采用蛋氨酸-胱氨酸缺乏饲料(MCD)喂养,简称A1组;另1组小鼠采用正常饲料喂养,简称A2组。12只非基因敲除小鼠也分为两组,每组6只。其中1组小鼠采用MCD喂养,简称B1组;另1组小鼠采用正常饲养喂养,简称B2组。两周后处死小鼠,测定其血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)、NLR家族Prin域3(NLRP3),并检测B细胞淋巴瘤因子-2(Bcl-2)、Bcl-2相关X蛋白(Bax蛋白)等表达。观察小鼠肝细胞凋亡情况。结果:HE染色与油红O染色显示,A1、A2组小鼠在2周时肝细胞变性程度明显弱于B1、B2组,P<0.05;A1、A2组小鼠肝细胞凋亡数量明显少于B1、B2组,P<0.05;A1、A2组小鼠血清ALT、AST、Bax水平低于B1、B2组,P<0.05,而Bcl-2水平高于B1、B2组,P<0.05;A1、A2组小鼠IL-1β、IL-18、NLRP3水平均低于B1、B2组,P<0.05。A1、B1组小鼠血清ALT、AST、分别高于A2、B2组,但A1组小鼠ALT、AST则低于B1组。结论:特异性敲除小鼠CD147基因,可以减弱小鼠肝组织脂肪变性,减少肝细胞凋亡(下调促凋亡分子Bax水平,上调抗凋亡分子Bcl-2水平)、降低细胞因子IL-1β、IL-18以及炎性小体NLRP3的表达,从而达到抑制NASH的目的。Objective:To study the possible mechanism of action specific knockout of CD147 to inhibit non-alcoholic steatohepatitis(NASH).Methods:Eight week old mice were selected,which were consistent with hepatocyte-specific knockout CD147 gene mice(Alb;Bsg flx-flx group) and their littermate control mice(Bsg flx-flx group),6 in each group.The corresponding methionine-cystine-deficient feed(MCD group) and control feed were administered separately(control group).Two weeks later,the mice were sacrificed and serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),interleukin-1β/(IL-1β-18),and NLR family pyrin domain 3(NLRP3) were measured,and the expression of B-cell lymphoma factor-2(Bcl-2),Bcl-2 related x protein(Bax) protein and apoptosis of mouse hepatocytes were detected.Results:HE staining and oil red O dye staining showed that the degree of steatosis at 2 weeks in the Alb;Bsg flx-flx group was significantly weaker than that in the Bsg flx-flx group.When MCD was induced,serum ALT and AST levels were higher in the MCD group than in the littermate control group(P<0.05),while serum ALT and AST levels in the Alb;Bsg flx-flx group were lower than those in the Bsg flx-flx group(P<0.05).When MCD was induced,the number of hepatocytes in the liver tissue of Alb;Bsg flx-flx group were significantly lower than that of Bsg flx-flx group(P<0.05);Alb;Bsg flx-flx group was lower than Bsg flx-flx group(P<0.05),while Alb;Bsg flx-flx group Bcl-2 level was higher than Bsg flx-flx group(P<0.05);Alb;Bsg flx-flx group IL-1β,IL-18 level,The NLRP3 level were lower than that of the Bsg flx-flx group(P<0.05).Conclusion:Hepatocyte knockout of CD147 in NASH can attenuate hepatic steatosis,hepatocyte apoptosis(down-regulation of pro-apoptotic Bax levels,up-regulation of anti-apoptotic Bcl-2 levels),cytokines IL-1β,IL-18,and Expression of inflammatory corpuscle NLRP3.

关 键 词：非酒精性脂肪性肝炎 CD147 特异性敲除 肝功能 炎性因子 

分 类 号：R575.1[医药卫生—消化系统]