国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2014年
15期
1167-1171
,共5页
肺成纤维细胞%钙离子%钙调磷酸酶
肺成纖維細胞%鈣離子%鈣調燐痠酶
폐성섬유세포%개리자%개조린산매
Pulmonary fibrosis%Ca2+%Calcineurin
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；Western blot 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；② TGF-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α-SMA 的表达；TGF-β1与环孢素 A共刺激1 h、4 h、24 h 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1 h 增加最为明显,而4 h 组和24 h 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。
目的：探討鈣離子-鈣調燐痠酶在大鼠肺成纖維細胞嚮肺肌成纖維細胞錶型轉化中的作用。方法體外分離培養大鼠肺成纖維細胞,免疫組織化學法檢測波形蛋白鑒定細胞；以 Fluo3-AM 為鈣離子熒光示蹤劑,激光共聚焦顯微鏡檢測細胞內基礎 Ca2+水平以及給予轉化生長因子β1(TGF-β1)刺激後細胞內鈣離子水平變化；Western blot 檢測各組細胞內α-SMA、p-Smad2的錶達。結果①大鼠肺成纖維細胞波形蛋白胞漿錶達呈暘性；② TGF-β1刺激後大鼠肺成纖維細胞胞內鈣離子水平升高；③TGF-β1刺激組,α-SMA 錶達明顯增加,加用環孢素 A 後不能抑製α-SMA 的錶達；TGF-β1與環孢素 A共刺激1 h、4 h、24 h 各組與 TGF-β1刺激組相比,p-Smad2水平均明顯增加,且以環孢素 A 作用1 h 增加最為明顯,而4 h 組和24 h 組比較差異無統計學意義。結論 TGF-β1刺激後大鼠肺成纖維細胞胞內鈣離子水平升高。鈣調燐痠酶可能參與肺成纖維細胞嚮肺肌成纖維細胞錶型轉化。
목적：탐토개리자-개조린산매재대서폐성섬유세포향폐기성섬유세포표형전화중적작용。방법체외분리배양대서폐성섬유세포,면역조직화학법검측파형단백감정세포；이 Fluo3-AM 위개리자형광시종제,격광공취초현미경검측세포내기출 Ca2+수평이급급여전화생장인자β1(TGF-β1)자격후세포내개리자수평변화；Western blot 검측각조세포내α-SMA、p-Smad2적표체。결과①대서폐성섬유세포파형단백포장표체정양성；② TGF-β1자격후대서폐성섬유세포포내개리자수평승고；③TGF-β1자격조,α-SMA 표체명현증가,가용배포소 A 후불능억제α-SMA 적표체；TGF-β1여배포소 A공자격1 h、4 h、24 h 각조여 TGF-β1자격조상비,p-Smad2수평균명현증가,차이배포소 A 작용1 h 증가최위명현,이4 h 조화24 h 조비교차이무통계학의의。결론 TGF-β1자격후대서폐성섬유세포포내개리자수평승고。개조린산매가능삼여폐성섬유세포향폐기성섬유세포표형전화。
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 ofα-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 ofα-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.
