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

作　　者：李婷婷 韩臻平 高立 王思懿 钟恺 黄萍 严洁萍[1,3] LI Tingting;HAN Zhenping;GAO Li;WANG Siyi;ZHONG Kai;HUANG Ping;YAN Jieping(Center for Clinical Pharmacy,Department of Pharmacy,Zhejiang Provincial People′s Hospital,People′s Hospital Affiliated to Hangzhou Medical College,Hangzhou 310014;Department of Pharmacology,College of Pharmaceutical Sciences,Zhejiang University of Technology,Hangzhou 310014;Key Laboratory of Endocrine Gland Diseases of Zhejiang Province,Hangzhou 310014,China)

机构地区：[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

出　　处：《基础医学与临床》2023年第11期1631-1635,共5页Basic and Clinical Medicine

基　　金：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)。

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。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.

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 

分 类 号：R364.4[医药卫生—病理学]