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*

doi:10.12047/j.cjap.6077.2021.086

中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (6): 589-593.doi: 10.12047/j.cjap.6077.2021.086

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^*

薛峰, 王辉, 许思多, 刘首挺, 龚永生, 范小芳^△

温州医科大学低氧医学研究所, 浙江温州 325035

收稿日期:2020-04-22 修回日期:2021-03-03 出版日期:2021-11-28 发布日期:2021-11-25
通讯作者: ^△Tel: 0577-86699521; E-mail: fxbwzmc@126.com
基金资助:
^*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)

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

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

Received:2020-04-22 Revised:2021-03-03 Online:2021-11-28 Published:2021-11-25

摘要/Abstract

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(10 μg/ ml,3、6、12、24 h)、LPS(10 μg/ ml,24 h)+LC(终浓度为2.5、5、10 μg/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。

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

Abstract: 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: ① Control group, ② LPS group (10 μg/ml, 3, 6, 12, 24 h), ③ LPS (10 μg/ml, 24 h)+LC (2.5, 5.0, 10 μg/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: ① Compared 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. ② Compared with LPS group (10 μg/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.

Key words: L-carnitine, lipopolysaccharide, mouse endothelial cells, autophagy, apoptosis

中图分类号:

R73-3

薛峰, 王辉, 许思多, 刘首挺, 龚永生, 范小芳. 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^*[J]. 中国应用生理学杂志, 2021, 37(6): 589-593.

XUE Feng, WANG Hui, XU Si-duo, LIU Shou-ting, GONG Yong-sheng, FAN Xiao-fang. Effects of L-carnitine on autophagy and apoptosis of mouse pulmonary microvascular endothelial cells induced by lipopolysaccharide[J]. CJAP, 2021, 37(6): 589-593.

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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

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