丙酮醛降低人脑微血管内皮细胞系hCMEC/D3细胞活力及线粒体膜电位

doi:10.16352/j.issn.1001-6325.2023.11.1631

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (11): 1631-1635.doi: 10.16352/j.issn.1001-6325.2023.11.1631

• 研究论文 • 下一篇

丙酮醛降低人脑微血管内皮细胞系hCMEC/D3细胞活力及线粒体膜电位

李婷婷¹, 韩臻平², 高立¹, 王思懿², 钟恺¹, 黄萍^1,3, 严洁萍^1,3*

1.浙江省人民医院杭州医学院附属人民医院临床药学中心药学部, 浙江杭州 310014;
2.浙江工业大学药学院药学系, 浙江杭州 310014;
3.浙江省内分泌腺体疾病诊疗重点实验室, 浙江杭州 310014

收稿日期:2023-02-07 修回日期:2023-07-18 出版日期:2023-11-05 发布日期:2023-10-30
通讯作者: ^*yanjieping@hmc.edu.cn
基金资助:
国家自然科学基金(81903597); 浙江省自然科学基金(LQ16H310003); 浙江省医药卫生科技项目(2021KY016, 2022KY061); 浙江省中医药科技计划(2023ZL015)

Methylglyoxal reduces cell viability and mitochondrion membrane potential of human brain microvascular endothelial cell line hCMEC/D3

LI Tingting¹, HAN Zhenping², GAO Li¹, WANG Siyi², ZHONG Kai¹, HUANG Ping^1,3, YAN Jieping^1,3*

1. Center for Clinical Pharmacy, Department of Pharmacy, Zhejiang Provincial People's Hospital, People's Hospital Affiliated to Hangzhou Medical College, Hangzhou 310014;
2. Department of Pharmacology, College of Pharmaceutical Sciences,Zhejiang University of Technology, Hangzhou 310014;
3. Key Laboratory of Endocrine Gland Diseases of Zhejiang Province, Hangzhou 310014, China

Received:2023-02-07 Revised:2023-07-18 Online:2023-11-05 Published:2023-10-30
Contact: ^*yanjieping@hmc.edu.cn

摘要/Abstract

摘要： 目的探讨丙酮醛(MGO)介导人脑微血管内皮细胞系hCMEC/D3的损伤作用及机制。方法不同浓度MGO(0.25、0.50、0.75、1.00和1.25 mmol/L)作用hCMEC/D3细胞24 h,用CCK-8法检测hCMEC/D3细胞活力;用试剂盒检测胞内乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)活性。线粒体膜电位检测试剂盒(JC-1)荧光探针检测线粒体膜电位水平,MitoSOX荧光探针观察线粒体活性氧(mROS)产生。结果 MGO组与对照组相比,hCMEC/D3细胞活力呈浓度依赖性下降(P＜0.01),细胞内LDH活性显著升高(P＜0.01),同时SOD活性降低(P＜0.01),差异有统计学意义。与对照组相比,MGO处理12 h后的hCMEC/D3细胞经JC-1染色红绿荧光比值降低,提示MGO诱导细胞线粒体膜电位下降。MitoSOX染色后,与对照组相比,MGO处理过的hCMEC/D3细胞的红色荧光表达数量增加,提示细胞内mROS过量生成。结论 MGO降低hCMEC/D3细胞活力及线粒体膜电位,并诱导胞内mROS过量生成。

关键词: 糖尿病脑血管并发症, 丙酮醛, 人脑微血管内皮细胞系, 膜电位, 线粒体活性氧

Abstract: Objective To explore the effects and mechanism of methylglyoxal (MGO) in human brain microvascular endothelial cell line (hCMEC/D3). Methods The hCMEC/D3 cells were incubated with MGO(0.25, 0.50, 0.75, 1.00 and 1.25 mmol/L) for 24 h. Cell viability was detected by CCK-8 assay. Cellular LDH and SOD activities were determined by commercially available kits. The change of mitochondrion membrane potential was assessed by JC-1 probe. MitoSOX probe was used to observe mitochondrial reactive oxygen species (mROS) release. Results Compared with control, cell viability of hCMEC/D3 treated with MGO decreased in a concentration-dependent manner (P＜0.01). Increased LDH activity and decreased SOD activity were observed with MGO exposure(P＜0.01). The decrease of mitochondrion membrane potential and the excessive increase of mROS were induced after MGO exposure for 12 h, which were detected by JC-1 or MitoSOX probe. Conclusions MGO reduces cell viability and mitochondria membrane potential in hCMEC/D3 cells with excessive mROS generation.

Key words: diabetic cerebral vascular complication, methylglyoxal, human brain microvascular endothelial cells line, membrane potential, mitochondrial reactive oxygen species

中图分类号:

R364.4

李婷婷, 韩臻平, 高立, 王思懿, 钟恺, 黄萍, 严洁萍. 丙酮醛降低人脑微血管内皮细胞系hCMEC/D3细胞活力及线粒体膜电位[J]. 基础医学与临床, 2023, 43(11): 1631-1635.

LI Tingting, HAN Zhenping, GAO Li, WANG Siyi, ZHONG Kai, HUANG Ping, YAN Jieping. Methylglyoxal reduces cell viability and mitochondrion membrane potential of human brain microvascular endothelial cell line hCMEC/D3[J]. Basic & Clinical Medicine, 2023, 43(11): 1631-1635.

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丙酮醛降低人脑微血管内皮细胞系hCMEC/D3细胞活力及线粒体膜电位

Methylglyoxal reduces cell viability and mitochondrion membrane potential of human brain microvascular endothelial cell line hCMEC/D3

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