钙离子-钙调磷酸酶对肺成纤维细胞表型转化的影响  

Effect of Ca2＋ -Calcineurin on fibroblast-to-myofibroblast differentiation

作　　者：董维维[1] 叶燕青[1] 王高明[1] 

机构地区：[1]武汉大学中南医院呼吸内科,430071

出　　处：《国际呼吸杂志》2014年第15期1167-1171,共5页International Journal of Respiration

基　　金：湖北省科技厅自然科学基金资助（2011CDB527）

摘　　要：目的探讨钙离子-钙调磷酸酶在大鼠肺成纤维细胞向肺肌成纤维细胞表型转化中的作用。方法体外分离培养大鼠肺成纤维细胞，免疫组织化学法检测波形蛋白鉴定细胞；以Fluo3-AM为钙离子荧光示踪剂，激光共聚焦显微镜检测细胞内基础Ca2＋水平以及给予转化生长因子β1（TGF-β1）刺激后细胞内钙离子水平变化；Westernblot检测各组细胞内α—sMA、P—Smad2的表达。结果①大鼠肺成纤维细胞波形蛋白胞浆表达呈阳性；②TGF-β1刺激后大鼠肺成纤维细胞胞内钙离子水平升高；③TGF—β1刺激组，α—SMA表达明显增加，加用环孢素A后不能抑制a-SMA的表达；TGF-β1与环孢素A共刺激1h、4h、24h各组与TGF-β1刺激组相比，p-Smad2水平均明显增加，且以环孢素A作用1h增加最为明显，而4h组和24h组比较差异无统计学意义。结论TGF-β1刺激后大鼠肺成纤维细胞胞内钙离子水平升高。钙调磷酸酶可能参与肺成纤维细胞向肺肌成纤维细胞表型转化。Objective To explore the effect of Ca2＋-Calcineurin on fibroblast-to-myofibroblast differentiation. Methods The lung fibroblasts were isolated from the lungs of SD rats at postnatal day 3. Use immunohistochemical technique to observe the expression of Vimentin. The fluorescence intensity of intracellular Ca2＋ was detected respectively with Fluo-3/AM loading by the laser confocal microscopy. The expression of α-SMA and p-Smad2 were measured by Western blotting. Results ①It can observe the expression of vimentin in primary lung fibroblasts. （2）TGF-β1 can increase of cytosolic Ca2＋ of primary lung fibroblasts.③TGF-β1 stimulated group was obvious increased the expression of α-SMA compared to Control group,CsA as the inhibitor of calcineurin can＇t inhibit the expression of α- SMA. TGF-β1 together with CsA stimulated of 1 h,4 h,24 h group were obvious increased the expression of p-Smad2 compared to TGF-β1 stimulated group,and 1 h group was more than the other groups. There were no differentiation between 4 h group and 24 h group. Conclusions There were changes of cytosolic Ca2＋ in rat lung fibroblasts by TGF-β1. CaN may take part in the process of fibroblast-to-myofibroblast differentiation.

关 键 词：肺成纤维细胞 钙离子 钙调磷酸酶 

分 类 号：R563.9[医药卫生—呼吸系统]