七氟醚对HT22细胞线粒体钙离子浓度影响的研究
Effect of sevoflurane on mitochondrial Ca2+concentration in HT22 cells
王晨 1李雨 2曹晨 1朱文竞 1闫慧宇 1程婵 1李璐 1王凯璇 1张研 2潘楚雄1
作者信息
- 1. 首都医科大学附属北京口腔医院麻醉科,北京 100050
- 2. 北京大学生命科学学院膜生物学国家重点实验室,北京 100871
- 折叠
摘要
目的 探究七氟醚对HT22细胞(小鼠海马神经元细胞系)线粒体Ca2+浓度和线粒体钙单向转运蛋白(MCU)的影响.方法 使用HT22细胞作为研究对象.HT22细胞采用随机数字表法分为对照组和七氟醚组,对照组细胞暴露于含对照溶剂的培养基,七氟醚组细胞置于含1 mmol/L七氟醚的培养基,两组细胞于37 ℃、5％CO2的培养箱中处理6 h.利用乳酸脱氢酶试剂盒评估细胞损伤情况,细胞增殖检测试剂盒评估细胞活力,利用细胞Ca2+探针Calbryte-520 AM和Rhod-2 AM评估细胞质和线粒体Ca2+浓度,利用线粒体探针PK Mito Deep Red进行线粒体定位,应用免疫印迹法检测MCU表达.结果 与对照组比较,七氟醚组HT22细胞活力降低(P＜0.05)、细胞损伤加重(P＜0.05)、细胞质和线粒体的Ca2+浓度明显升高(P＜0.05),但MCU表达水平差异无统计学意义(P＞0.05).结论 七氟醚可以引起线粒体钙超载,但不影响MCU表达水平.
Abstract
Objective To investigate the effect of sevoflurane on mitochondrial Ca2+concentration and mitochondrial calcium uniporter(MCU)in HT22 cells(mouse hippocampal neuronal cell line).Methods HT22 cells were used for the study.According to the random number table method,HT22 cells were divided into two groups:a control group and a sevoflurane group.Cells in the control group were exposed to the medium containing control solvent,while the sevoflurane group was placed in medium containing 1 mmol/L sevoflurane.All the cells were cultivated an incubator at 37 ℃ and 5％CO2 for 6 h.The cell damage was detected by lactate dehydroge-nase kit.The cell viability was assessed by cell proliferation assay kit.The cytoplasmic-and mitochondrial Ca2+concentrations were as-sessed using the cellular Ca2+probes Calbryte-520 AM and Rhod-2 AM.Mitochondrial localization was performed using the mitochon-drial probe PK Mito Deep Red.The expression of MCU was analyzed by Western blot.Results Compared with the control group,the sevoflurane group showed reduced HT22 cell viability(P＜0.05),aggravated cell damage(P＜0.05),and significantly increased Ca2+concentrations in the cytoplasm and mitochondria(P＜0.05),but no statistical difference was found in MCU expression(P＞0.05).Con-clusion Sevoflurane can cause mitochondrial calcium overload,but does not affect the MCU expression.
关键词
HT22细胞/七氟醚/线粒体/钙超载/线粒体钙单向转运蛋白Key words
HT22 cell/Sevoflurane/Mitochondria/Calcium overloading/Mitochondrial calcium uniporter引用本文复制引用
出版年
2024
NETL
NSTL
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