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论文摘要

毒胡萝卜素对小鼠肺成纤维细胞内钙离子水平及caspase-3蛋白表达的影响

Effects of Thapsigargin on Intracellular Ca2+ and Caspase-3 Protein Expression in Mouse Lung Fibroblast

作者:闫宇, 肖坤, 梁晓飞等

Author:YAN Yu, XIAO Kun, LIANG Xiao-fei. et al

收稿日期:          年卷(期)页码:2018,49(3):380-383

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:肺成纤维细胞 毒胡萝卜素 钙离子 含半胱氨酸的天冬氨酸蛋白水解酶-3

Key words:Lung fibroblasts Thapsigargin Calcium caspase-3

基金项目:

中文摘要

目的 探讨钙泵抑制剂毒胡萝卜素(thapsigargin,TG)对体外小鼠肺成纤维细胞内钙离子(Ca2+)水平及凋亡相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)表达的影响。方法 小鼠肺成纤维细胞(L929)分为3组:空白对照组(不做处理)、转化生长因子β1(TGF-β1)组、TG组,后两组均用5 ng/mL TGF-β1诱导24 h,TG组在诱导后再用4 μmol/L TG作用24 h。细胞处理48 h后,分别收集各组细胞,采用透射电子显微镜观察细胞超微结构变化,激光共聚焦显微镜观察各组细胞内Ca2+水平,免疫细胞化学方法检测caspase-3蛋白表达。结果 透射电子显微镜示,空白对照组细胞凋亡较少,TGF-β1组无明显凋亡细胞,TG组有大量成纤维细胞凋亡。TGF-β1组Ca2+水平和caspase-3蛋白水平均低于空白对照组(P2+水平和caspase-3蛋白水平均较TGF-β1组、空白对照组升高(P

英文摘要

Objective To explore the effects of calcium pump inhibitor thapsigargin (TG) on intracellular Ca2+ and the expression of caspase-3 protein in mouse lung fibroblast. Methods Mice lung fibroblast cells were divided into three groups: the blank control group, transforming growth factor-β1(TGF-β1) group and TG treated group. The cells were induced by 5 ng/mL TGF-β1 for 24 h in TGF-β1 group and TG group, 4 μmol/L TG was added in TG group for 24 h. After 48 h, the cells were collected, and the cell structure was observed by transmission electron microscope, intracellular Ca2+ level was detected with laser confocal microscope, the protein expression of caspase-3 was examined using immunohistochemistry method. Results Transmission electron microscopy showed that the cells in the blank control group had obvious nucleolus, complete organelles and less apoptosis. In TGF-β1 group, the cell morphology was intact, chromatin was evenly distributed, and no apoptotic cells were found. In TG group, there were a large number of apoptosis of fibroblasts, chromatin clumps in nuclei and a small amount of collagen fibers. The level of Ca2+ in TGF-β1 group was significantly lower than that in control group (PP1 group were significantly lower than that in blank control group (PP

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