解整合素-金属蛋白酶8对急性酒精性肝损伤小鼠中肝细胞增殖与凋亡的调控作用  

Regulation of proliferation and apoptosis of hepatocytes in mice with acute alcoholic liver injury by ADAM8

作　　者：崔钦奕 杨孟利 李三强[1] 齐婧含 冯家阳 张凯杰 CUI Qinyi;YANG Mengli;LI Sanqiang;QI Jinghan;FENG Jiayang;ZHANG Kaijie(Key Laboratory of Molecular Medicine for Liver Injury and Repair,College of Basic Medicine and Law,Henan University of Science and Technology,Luoyang 471003,China)

机构地区：[1]河南科技大学基础医学与法医学院肝脏损伤与修复分子医学重点实验室,河南洛阳471003

出　　处：《胃肠病学和肝病学杂志》2024年第8期1005-1009,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然基金资助项目(82170606);河南省高等学校重点科研项目计划基础研究专项(23ZX006)。

摘　　要：目的 探讨解整合素-金属蛋白酶8(A disintegrin and metalloprotease 8,ADAM8)对急性酒精性肝损伤小鼠中肝细胞增殖与凋亡的影响。方法 选择6~8周龄的KM小鼠30只,随机分为5组:正常对照组(n=6)、模型组(n=6)、ADAM8-sgRNA1组(n=6)、ADAM8-sgRNA2组(n=6)、ADAM8-sgRNA3组(n=6)。正常对照组小鼠不做任何处理,质粒组小鼠分别尾静脉注射3种质粒,模型组小鼠尾静脉注射等量生理盐水;将模型组和质粒组小鼠常规喂养3 d,通过一次经口灌胃50%酒精14 mL/kg诱导小鼠急性肝损伤。检测各组小鼠AST、ALT水平,HE染色观察其病理学变化,PAS染色观察肝组织糖原变化;Western blotting检测ADAM8、TNF-α、增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)、BCL2关联X蛋白(BCL2-associated X protein, Bax)的表达水平。结果 与模型组相比,ADAM8-sgRNA2组ADAM8的表达水平明显降低(P<0.05);与正常对照组相比,模型组PCNA的表达水平明显降低(P<0.001),TNF-α、Bax的表达水平明显升高(P<0.01);与模型组相比,ADAM8-sgRNA2组PCNA表达水平显著升高(P<0.001),TNF-α、Bax表达水平显著下降(P<0.01)。与正常对照组相比,模型组AST、ALT指标显著升高(P<0.05);与模型组相比,ADAM8-sgRNA2组AST、ALT指标显著下降(P<0.05)。HE染色和PAS染色结果显示:与正常对照组相比,模型组肝损伤明显,坏死积分显著升高(P<0.05),糖原含量显著减少(P<0.001);与模型组相比,ADAM8-sgRNA2组坏死积分明显降低(P<0.05),ADAM8-sgRNA2组糖原含量明显上升(P<0.001)。结论 ADAM8过表达可抑制急性酒精性肝损伤小鼠中肝细胞的增殖并促进肝细胞的凋亡。Objective To investigate the effects of ADAM8 on hepatocyte proliferation and apoptosis in mice with acute alcoholic liver injury.Methods Thirty KM mice aged 6-8 weeks were randomly divided into 5 groups:normal control group(n=6),model group(n=6),ADAM8-sgRNA1 group(n=6),ADAM8-sgRNA2 group(n=6),and ADAM8-sgRNA3 group(n=6).Mice in normal control group did not receive any treatment,mice in plasmid group were injected with 3 plasmids respectively through tail vein,and mice in model group were injected with the same amount of normal saline through tail vein.Mice in model group and plasmid group were routinely fed for 3 days,and acute liver injury was induced by oral gavage of 14 mL/kg 50%alcohol once.The serum was separated for the determination of AST and ALT.The liver of the mice was stained with HE to observe the histopathological changes and the liver glycogen changes were stained with PAS.The expression levels of ADAM8,TNF-α,PCNA and Bax were detected by Western blotting.Results The expression level of ADAM8 in ADAM8-sRNA2 group was significantly decreased compared with model group(P<0.05).Compared with the normal control group,the expression level of PCNA in the model group was significantly decreased(P<0.001),and the expression levels of TNF-αand Bax were significantly increased(P<0.01).Compared with model group,the expression levels of PCNA in ADAM8-sgRNA2 group were significantly increased(P<0.001),and the expression levels of TNF-αand Bax were significantly decreased(P<0.01 or P<0.001).Compared with normal control group,AST and ALT indexes in model group were significantly increased(P<0.05).Compared with model group,AST and ALT indexes of ADAM8-sgRNA2 group were significantly decreased.The results of HE staining and PAS staining showed that compared with the normal control group,the liver injury was obvious in the model group,the necrosis score was significantly increased(P<0.05),and the glycogen content was significantly decreased(P<0.001).Compared with model group,necrosis score in ADAM8-sgRNA2 group wa

关 键 词：急性酒精性肝损伤 解整合素-金属蛋白酶8 CRISPR/Cas9 增殖 凋亡 

分 类 号：R575[医药卫生—消化系统]