左旋卡尼汀对脂多糖诱导小鼠肺微血管内皮细胞自噬和凋亡的影响  

Effects of L-carnitine on autophagy and apoptosis of mouse pulmonary microvascular endothelial cells induced by lipopolysaccharide

作　　者：薛峰[1] 王辉[1] 许思多 刘首挺 龚永生[1] 范小芳[1] XUE Feng;WANG Hui;XU Si-duo;LIU Shou-ting;GONG Yong-sheng;FAN Xiao-fang(Institute of Hypoxia Medicine,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学低氧医学研究所,浙江温州325035

出　　处：《中国应用生理学杂志》2021年第6期589-593,共5页Chinese Journal of Applied Physiology

基　　金：浙江省自然科学研究项目(LY18H010007);温州市科技计划项目(2016Y0831);国家自然科学基金青年基金项目(81600041)。

摘　　要：目的:探讨左旋卡尼汀(LC)对脂多糖(LPS)损伤的小鼠肺微血管内皮细胞(PMVECs)的保护作用及自噬、凋亡的影响。方法:采用体外培养的小鼠PMVECs,分为对照组(Control组)、LPS组(10μg/ml,3、6、12、24 h)、LPS(10μg/ml,24 h)+LC(终浓度为2.5、5、10μg/ml)(LC组)。Annexin V-FITC/PI双标记法检测细胞凋亡,细胞免疫荧光染色法检测自噬小体,Western blot法检测自噬相关蛋白LC3及凋亡蛋白Caspase-3的含量,CCK-8法检测细胞活力。结果:①与Control组比较,LPS 6 h、12 h、24 h组PMVECs细胞活力显著受到抑制,细胞凋亡率、自噬蛋白LC3Ⅱ表达显著增高(P均<0.01),LC3蛋白阳性表达。②与LPS 24 h组比较,各浓度LC组PMVECs细胞活力显著提高、自噬蛋白LC3II表达水平显著升高(P均<0.01),而PMVECs凋亡率和凋亡蛋白Caspase-3表达水平均明显降低(P<0.05)。结论:LC具有提高LPS刺激的小鼠PMVECs活性、促进PMVECs自噬、抑制凋亡的作用。Objective:To investigate the protective effects of L-carnitine(LC)on lipopolysaccharide(LPS)-injured mouse pulmonary microvascular endothelial cells(PMVECs)and its effects on autophagy and apoptosis.Methods:Cultured mouse PMVECs were divided into three groups:①Control group,②LPS group(10μg/ml,3,6,12,24 h),③LPS(10μg/ml,24 h)+LC(2.5,5.0,10μg/ml)(LPS+LC)group.PMVECs apoptosis was examined by Annexin V-FITC/PI double labeling method.Autophagosome was detected by immunofluorescence staining.Levels of autophagy-related protein LC3 and apoptosis-related protein caspase-3 were detected by Western blot.PMVECs viability was measured by CCK-8.Results:①Compared with the control group,LPS treatment inhibited the PMVECs viability significantly,whereas the apoptosis rate and the expression of autophagy protein LC3 II were markedly increased after LPS treatment for 6 h,12 h and 24 h.②Compared with LPS group(10μg/ml,24 h),the PMVECs viability,levels of autophagy protein LC3 II and caspase-3 protein expression as well as apoptosis rate in LPS+LC group were increased significantly.Conclusion:LC can increase the activity of PMVECs injuried by LPS,promote autophagy and inhibit apoptosis of PMVECs.

关 键 词：左旋卡尼汀 脂多糖 小鼠肺微血管内皮细胞 自噬 凋亡 

分 类 号：R73-3[医药卫生—肿瘤]