肺成纤维细胞在博来霉素致肺纤维化中的作用机制  

Pulmonary fibroblast cells play an important role of mechanism in causing pulmonary fibrosis induced by bleomycin

作　　者：何爱明[1] 金美芳[1] 叶瑞洪[1] 

机构地区：[1]福建师范大学福清分校生化系,350300

出　　处：《心肺血管病杂志》2015年第11期863-866,共4页Journal of Cardiovascular and Pulmonary Diseases

摘　　要：目的:探讨肺成纤维细胞(PFC)在博来霉素(BLM)致肺纤维化中的作用机制。方法:采用差时贴壁法培养与纯化PFC,传至第3代备用。分为Control组:细胞用正常的培养基培养;BLM组:依次分为0.01mol/L BLM组、0.03mol/L BLM组、0.1mol/L BLM组、1mol/L BLM组及10mol/L BLM,每组每孔细胞分别加上述浓度的BLM,5×104个PFC/孔(12孔板),干预24小时。TGF-β1组:每个孔用2ug/L TGF-β1,5×104个PFC/孔(12孔板),干预24小时。免疫细胞化学法鉴定PFC与肺肌成纤维细胞(PMFC)。采用Real-time PCR及Western-blot技术检测Ⅰ、Ⅲ型胶原mRNA及α-SMA mRNA表达与α-SMA蛋白水平。Brd U法检测细胞增殖。结果:BLM可剂量依赖性地抑制细胞增殖,BLM浓度从0.1mol/L始抑制PFC的增殖较对照组(P<0.05)。0.1mol/L BLM干预24小时后,α-SMA免疫细胞化学染色鉴定呈阳性,α-SMA mRNA的较对照组表达增加(P<0.05);Ⅰ、Ⅲ型胶原mRNA的表达较对照组显著增加(P<0.01);0.1mol/L BLM能明显升高α—SMA的蛋白表达水平。结论:PFC转化成PMFC且显著增加Ⅰ、Ⅲ型胶原的表达,成为BLM诱导肺纤维化中的重要作用机制之一。Objective： To study on pulmonary fibroblast cells（ PFC） playing an important role of mechanism in causing pulmonary fibrosis induced by Bleomycin（ BLM）. Methods： Using differential adherent method to cultivate and purify PFC and generating the third to use. The groups were divided into control,0. 01 mol / L BLM,0. 03 mol / L BLM,0. 1mol / L BLM,1mol / L BLM,10 mol / L BLM,2ng / m L TGF-β1. 5× 104 PFC were respectively treated with above drug concentration for 24 hours. PFC and PMFC were identified by immunocytochemistry. Cell proliferation was measured by Brd U marking. The expression of Ⅰ、Ⅲ collagen and α—SMA were analysed by real-time PCR or Western blot. Results： BLM inhibited cell proliferation by dose—dependent from 0. 1 mol / L（ P 0. 05 vs Control）. Immunocytochemistry staining identification of α-SMA was positive by administrating 0. 1 mol / L BLM for 24 hours. Up-regulated expression of α-SMA,and Ⅰand Ⅲ collagen in induced by BLM. Conclusion： PFC converting into PMFC and up-regulated expression of α-SMA,and Ⅰand Ⅲcollagen induced by BLM become one of the important mechanism of pulmonary fibrosis.

关 键 词：肺成纤维细胞 肺肌成纤维细胞 博来霉素 肺纤维化 

分 类 号：R54[医药卫生—心血管疾病]