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 Ca^(2+) concentration in HT22 cells

作　　者：王晨 李雨 曹晨 朱文竞 闫慧宇 程婵 李璐 王凯璇 张研[2] 潘楚雄 Wang Chen;Li Yu;Cao Chen;Zhu Wenjing;Yan Huiyu;Cheng Chan;Li Lu;Wang Kaixuan;Zhang Yan;Pan Chuxiong(Department of Anesthesiology,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China;State Key Laboratory of Membrane Biology,College of Life Sciences,Peking University,Beijing 100871,China)

机构地区：[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

出　　处：《国际麻醉学与复苏杂志》2024年第7期673-677,共5页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金(81671044)。

摘　　要：目的探究七氟醚对HT_(2)2细胞(小鼠海马神经元细胞系)线粒体Ca^(2+)浓度和线粒体钙单向转运蛋白(MCU)的影响。方法使用HT_(2)2细胞作为研究对象。HT_(2)2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%CO_(2)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Ca^(2+)探针Calbryte-520 AM和Rhod-2 AM评估细胞质和线粒体Ca^(2+)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HT_(2)2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Ca^(2+)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。Objective To investigate the effect of sevoflurane on mitochondrial Ca^(2+)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%CO_(2)for 6 h.The cell damage was detected by lactate dehydrogenase kit.The cell viability was assessed by cell proliferation assay kit.The cytoplasmic and mitochondrial Ca^(2+)concentrations were assessed using the cellular Ca^(2+)probes Calbryte-520 AM and Rhod-2 AM.Mitochondrial localization was performed using the mitochondrial 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 Ca^(2+)concentrations in the cytoplasm and mitochondria(P<0.05),but no statistical difference was found in MCU expression(P>0.05).ConclusionSevoflurane 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 

分 类 号：R614[医药卫生—麻醉学]