靶向沉默CaMKK-AMPK通路对NEFAs介导肝脂损伤的调控机制  

Regulatory mechanism of targeted silencing of CaMKK-AMPK pathway on NEFAs mediated liver lipid injury

作　　者：王爽[1] 任大禹 张睿锴 杨威[1] 张冰冰[2] 董志昊 赵莹莹 麻芯茹 徐闯[1] WANG Shuang;REN Dayu;ZHANG Ruikai;YANG Wei;ZHANG Bingbing;DONG Zhihao;ZHAO Yingying;MA Xinru;XU Chuang(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China;College of Life Science and Technology,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]黑龙江八一农垦大学生命科学技术学院,黑龙江大庆163319

出　　处：《中国兽医学报》2021年第2期322-326,共5页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2016YFD0501206)。

摘　　要：为探究Ca^(2+)/钙调素依赖性蛋白激酶(CaMKK)-AMP依赖蛋白激酶(AMPK)对非酯化脂肪酸(NEFAs)介导的AML12细胞肝脂损伤的调控机制,本试验通过添加高浓度的NEFAs刺激AML12细胞,RNA靶向沉默CaMKK,检测不同分组细胞内AMPK及固醇调节原件结合蛋白(SREBP1)的蛋白表达水平以及利用流式细胞术检测活性氧(ROS)的产生水平,同时利用激光共聚焦检测细胞内脂滴变化情况。结果显示,siCaMKK+NEFAs组AMPK蛋白表达水平较空白对照组及NEFAs组均显著降低,而SREBP1蛋白表达水平、ROS水平及脂滴数量均显著高于其他各组。结果表明,CaMKK通过AMPK信号通路调控SREBP1及线粒体脂质氧化可调节NEFAs诱导的肝脂损伤过程。To explore the regulatory mechanism of Ca^(2+)/calmodulin-dependent protein kinase(CaMKK)and AMP-dependent protein kinase(AMPK)on non-esterified fatty acids(NEFAs)mediated liver lipid injury in AML12 cells,AML12 cells were firstly treated with high concentrations of NEFAs,and then the expression of CaMKK was silenced by RNAi,followed by detection of protein expression level of AMPK and sterol regulatory element binding protein 1(SREBP1)in different groups of cells.The expression level of ROS was measured by flow cytometry assay,and the change of lipid droplets in cells were detected by laser confocal microscopy.The results showed that the expression level of AMPK protein in siCaMKK+NEFAs group was significantly lower than that in control group and NEFAs group,while the expression level of SREBP1 protein,ROS level and the number of lipid droplets were significantly higher than those in other groups.The results showed that CaMKK regulates SREBP1 and mitochondrial lipid oxidation via AMPK signaling pathway,which can regulate NEFAs induced liver lipid injury.

关 键 词：AML12 非酯化脂肪酸(NEFAs) CaMKK ROS 脂质沉积 

分 类 号：S852.3[农业科学—基础兽医学]