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闫宇, 肖坤, 梁晓飞等. 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[J]. koko体育app 学报（医学版）, 2018, 49(3): 380-383.
引用本文: 闫宇, 肖坤, 梁晓飞等. 毒胡红萝卜素对小鼠肺成合成纤维癌细胞内钙亚铁离子标准及caspase-3蛋白质传达的直接影响[J]. 江苏社会学报（医美版）, 2018, 49(3): 380-383.
YAN Yu, XIAO Kun, LIANG Xiao-fei. et al. Effects of Thapsigargin on Intracellular Ca2+ and Caspase-3 Protein Expression in Mouse Lung Fibroblast[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(3): 380-383.
Citation: YAN Yu, XIAO Kun, LIANG Xiao-fei. et al. Effects of Thapsigargin on Intracellular Ca2+ and Caspase-3 Protein Expression in Mouse Lung Fibroblast[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(3): 380-383.

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

  • 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（P<0.05），TG组细胞内Ca2+水平和caspase-3蛋白水平均较TGF-β1组、空白对照组升高（P<0.05）。结论 TG可使小鼠肺成纤维细胞细胞内钙稳态失衡，增加caspase-3蛋白表达，促进细胞凋亡。  
    Abstract: 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 (P<0.05), and which in TG group was significantly higher (P<0.05 ). The protein expression of caspase-3 in TGF-β1 group were significantly lower than that in blank control group (P<0.05), which in TG group increased obviously (P<0.05).Conclusion TG could cause intracellular calcium dysregulation in mouse lung fibroblasts, increase caspase-3 protein expression and promote cell apoptosis.  
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