线粒体Ca^2+浓度在高浓度氧致大鼠肺泡Ⅱ型上皮细胞凋亡中的作用  

Role of mitochondrial Ca^2+ concentration in hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells of rats

作　　者：程晓彤 李玉兰[1] 赵彦琴 李春兰 任程晖 Cheng Xiaotong;Li Yulan;Zhao Yanqin;Li Chunlan;Ren Chenghui(Department of Anesthesiology,the First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一医院麻醉科,730000

出　　处：《中华麻醉学杂志》2020年第9期1093-1095,共3页Chinese Journal of Anesthesiology

摘　　要：目的探讨线粒体Ca2+浓度在高浓度氧致大鼠肺泡Ⅱ型上皮细胞凋亡中的作用。方法取对数生长期的大鼠肺泡Ⅱ型上皮细胞,采用随机数字表法分为3组(n=6):对照组(C组)、高浓度氧组(H组)和高浓度氧+钌红组(HR组)。C组置于常规培养箱(5%CO2-95%空气)中培养4 h;HR组加入线粒体钙单向转运体抑制剂钌红2μmol/L,然后H组和HR组置于高浓度氧密闭箱(90%O2-5%CO2-5%空气)中培养4 h。通过倒置相差显微镜观察细胞形态学;采用流式细胞术测定细胞凋亡率;采用多功能酶标仪检测线粒体内Ca2+荧光强度,并计算Ca2+浓度。结果与C组比较,H组细胞凋亡率和线粒体Ca2+浓度升高,HR组线粒体Ca2+浓度降低(P<0.05),细胞凋亡率差异无统计学意义(P>0.05)。与H组比较,HR组细胞凋亡率和线粒体Ca2+浓度降低(P<0.05)。H组细胞呈现早期凋亡形态学表现(细胞皱缩呈球形、透亮度降低、与周围细胞脱离),HR组较H组明显改善。结论线粒体Ca2+浓度升高参与了高浓度氧致大鼠肺泡Ⅱ型上皮细胞凋亡。Objective To investigate the role of mitochondrial Ca2+concentration in hyperoxia-induced apoptosis in alveolar typeⅡepithelial cells of rats.Methods Alveolar typeⅡepithelial cells in the logarithmic growth phase were divided into 3 groups(n=6 each)using a random number table method:control group(group C),hyperoxia group(group H),and hyperoxia plus ruthenium red group(group HR).Cells in group C were cultured for 4 h in a conventional incubator(5%CO2-95%air).Cells in group HR were pretreated with mitochondrial calcium uniporter inhibitor ruthenium red 2μmol/L.Then H and HR groups were cultured for 4 h in a sealed oxygen chamber(90%O2-5%CO2-5%air)with high-oxygen concentration.Cell morphology was observed with the inverted phase contrast microscope,and the apoptosis rate of cells was determined by flow cytometry.The intensity of Ca2+fluorescence in mitochondria was measured by multiscan spectrum,and the concentration of Ca2+was calculated.Results Compared with group C,the apoptosis rates and mitochondrial Ca2+concentrations were significantly increased in group H,and the mitochondrial Ca2+concentration was significantly decreased(P<0.05),and no significant change was found in apoptosis rates in group HR(P>0.05).Compared with group H,the apoptosis rates and mitochondrial Ca2+concentrations were significantly decreased in group HR(P<0.05).Cells showed morphological manifestations of early apoptosis(cells shriveled into a spherical shape with reduced transmittance and detachment from surrounding cells)in group H,and the manifestation was significantly improved in group HR compared with group H.Conclusion Increased mitochondrial Ca2+concentration is involved in the hyperoxia-induced apoptosis in alveolar typeⅡepithelial cells of rats.

关 键 词：氧 肺泡 上皮细胞 细胞凋亡 线粒体 钙 

分 类 号：R619[医药卫生—外科学] R563[医药卫生—临床医学]