肝特异性敲除AMPKα对小鼠糖代谢基因的影响  

Effects of liver-specifie knockout of AMPKa on glycometabolism genes in mice

作　　者：张慧明 高倩 胡艳波 张笑茹 张钟月 杨艳 高峰 王敏杰 ZHANG Hui-ming;GAO Qian;HU Yan-bo;ZHANG Xiao-ru;ZHANG Zhong-yue;YANG Yan;GAO Feng;WANG Min-jie(School of Basic Medical Sciences,Inner Mongolia Medecial University,Hohhot 010110,Inner Mongolia Autonomous Region,China;College of Pharmacy,Inner Mongolia Medecial University,Hohhot 010110,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古医科大学基础医学院,内蒙古自治区呼和浩特010110 [2]内蒙古医科大学药学院,内蒙古自治区呼和浩特010110

出　　处：《中国临床药理学杂志》2024年第8期1184-1188,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81703511);内蒙古自治区草原英才青年创新创业计划(一层次)基金资助项目(11150000011512968L/2023-01172);内蒙古自治区自然科学基金面上基金资助项目(2023LHMS08068)。

摘　　要：目的 探究肝特异性敲除AMP活化蛋白激酶α(AMPKα)对小鼠胰腺功能和糖代谢相关基因的影响。方法 将AMPKα1/α2flox/flox小鼠分为空白组(给予普通饮食)和模型组(给予60%高脂胆碱缺乏饲料饲养),每组8只;另取AMPKα1/α2flox/flox/Alb-Cre+小鼠8只,设为敲除组,给予60%高脂胆碱缺乏饲料饲养,造模16周。用试剂盒检测小鼠的血脂水平和肝功能。用转录组测序检测小鼠胰腺中的差异表达基因。用实时荧光定量聚合酶链反应和蛋白质印迹法检测小鼠差异基因的表达情况。结果 空白组、模型组和敲除组的三酰甘油分别为(0.94±0.11)、(0.71±0.14)和(1.05±0.17)mmol·L^(-1),高密度脂蛋白分别为(1.62±0.07)、(0.44±0.08)和(0.90±0.06)mmol·L^(-1),谷草转氨酶分别为(7.02±5.87)、(15.60±3.15)和(22.70±2.14)U·L^(-1),谷丙转氨酶分别为(14.56±11.55)、(48.64±15.84)和(75.40±11.96)U·L^(-1),磷酸烯醇式丙酮酸羧激酶1(PCK1)mRNA分别为1.00±0、1.37±0.25、0.31±0.18,PCK1蛋白相对表达水平分别为0.77±0.27、1.23±0.43和0.51±0.40。敲除组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论 PCK1基因可能是介导肝AMPKα影响胰岛功能和糖代谢的关键基因。Objective To investigate the effects of liver-specific knockout of adenosine 5'-monophosphate-activated protein kinaseα(AMPKα)on pancreatic function and glucose metabolism-related genes in mice.Methods AMPKα_1/α_2~(flox/flox)mice were divided into blank group(common feed)and model group(60%high fat choline deficiency feet)with eight mice in each group,and another 8 AMPKα_1/α_2~(flox/flox/)Alb-Cre~+mice were divided into the knockout group(60%high fat choline deficiency feet).The kit detected the levels of blood lipids and liver function indexes.The differential genes in the mouse pancreas were detected by transcriptome sequencing.The expression of differential genes in mice was detected by real-time fluorescence quantitative polymerase chain reaction and Western blotting.Results The levels of triglyceride in the blank group,model group and knockout group were(0.94±0.11),(0.71±0.14)and(1.05±0.17)mmol·L^(-1);the levels of triglyceride and high-density lipoprotein were(1.62±0.07),(0.44±0.08)and(0.90±0.06)mmol·L^(-1);the levels of glutamic oxaloacetic transaminase were(7.02±5.87),(15.60±3.15)and(22.70±2.14)U·L^(-1);the levels of glutamic pyruvic transaminase were(14.56±11.55),(48.64±15.84)and(75.40±11.96)U·L^(-1);the expression levels of phosphoenolpyruvate carboxykinase 1(PCK1)mRNA were 1.00±0,1.37±0.25 and 0.31±0.18;the relative expression levels of PCK1 protein were 0.77±0.27,1.23±0.43 and 0.51±0.40,respectively.Significant differences existed in the above indexes between the knockout group and the model group(all P<0.05).Conclusion PCK1 gene may be an essential gene mediating the effect of liver AMPKαon islet function.

关 键 词：非酒精性脂肪性肝病 AMP活化蛋白激酶α 胰腺 糖代谢 磷酸烯醇式丙酮酸羧激酶1 

分 类 号：R97[医药卫生—药品]