TGF-β3对辐射致肺上皮细胞损伤的保护作用及其机制研究  

Protective effect and mechanism of TGF-β3 on lung epithelial cell injury induced by radiation

作　　者：鄢成名 杨陟华 王易龙 耿爽 刘奔波 王志鑫 李倩 王美玉[1,3] 郭浩鑫 朱茂祥 YAN Cheng-ming;YANG Zhi-hua;WANG Yi-long;GENG Shuang;LIU Ben-bo;WANG Zhi-xin;LI Qian;WANG Mei-yu;GUO Hao-xin;ZHU Mao-xiang(Beijing Key Laboratory of Radiobiology,Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Graduate School,University of South China,Hengyang,Hunan 421001,China;School of Life Sciences,Hebei University,Baoding,Hebei 071000,China)

机构地区：[1]军事科学院军事医学研究院辐射医学研究所,北京市放射生物学重点实验室,北京100850 [2]南华大学研究生院,湖南衡阳421001 [3]河北大学生命科学学院,河北保定071000

出　　处：《军事医学》2023年第1期20-25,共6页Military Medical Sciences

基　　金：国家自然科学基金(81673095)。

摘　　要：目的探索转化生长因子-β3(TGF-β3)在辐射致肺上皮细胞损伤中的作用及其可能机制,为放射性肺损伤的防治提供实验依据。方法设计合成并筛选Ⅱ型转化生长因子-β受体(TGFBR2)的小干扰RNA(siRNA),用于细胞TGFBR2干涉;将体外培养的人支气管上皮细胞(Beas-2B细胞)及其TGFBR2干涉细胞随机分为对照组、5 ng/ml TGF-β3处理组(TGF-β3组)、6 Gy60Co-γ射线单次照射组(6 Gy组)、6 Gy60Co-γ射线单次照射+5 ng/ml TGF-β3处理组(6 Gy+TGF-β3组),采用实时荧光定量PCR(RT-qPCR)检测各组胶原蛋白和TGFBR2的表达,RT-qPCR和Western印迹检测上皮间质转化(EMT)相关标志物平滑肌肌动蛋白(α-SMA)的表达;流式细胞术分析细胞凋亡和细胞周期。结果(1)TGF-β3对辐射诱发的细胞胶原蛋白表达增高有明显抑制作用,并可降低辐射所致细胞EMT相关标志物α-SMA基因和蛋白水平的异常高表达,有效保护辐射所致肺组织细胞纤维化改变;(2)TGF-β3可抑制辐射诱导的细胞TGFBR2异常高表达,并对辐射诱发的细胞凋亡和细胞周期G2/M期阻滞均有明显保护作用;(3)对细胞TGFBR2干涉后,TGF-β3对辐射所致细胞凋亡和细胞周期G2/M期阻滞的保护作用丧失,抑制辐射所致肺上皮细胞损伤。结论TGF-β3通过TGFBR2保护辐射损伤的肺上皮细胞。Objective To explore the role and possible mechanism of transforming growth factor-β3(TGF-β3)in radiation-induced lung epithelial cell injury,and to provide data for the prevention and treatment of radiation-caused lung injury.Methods The typeⅡtransforming growth factor-βreceptor(TGFBR2)siRNA was designed,synthesized and screened for cell TGFBR2 interference.Human bronchial epithelial cells(Beas-2B cells)and their TGFBR2 interference cells were randomly divided into the control group,5 ng/ml TGF-β3 acting group(TGF-β3 group),6 Gy60Co-γ-ray irradiation group(6 Gy group),and 6 Gy60Co-γ-ray irradiation+5 ng/ml TGF-β3 action group(6 Gy+TGF-β3 group).The expressions of collagens and TGFBR2 in each group were detected by real-time fluorescence quantitative PCR(RT-qPCR),and the expression of smooth muscle actin(α-SMA),a marker of epithelial-mesenchymal transition(EMT),was detected by RT-qPCR and Western blotting.Cell apoptosis and cell cycle were analyzed by flow cytometry.Results(1)TGF-β3 could significantly inhibit the expressions of collagens induced by radiation and reduce the abnormally high expressions ofα-SMA gene and protein,which was a marker of EMT induced by radiation,suggesting that TGF-β3 could effectively protect against lung cell fibrosis induced by radiation.(2)TGF-β3 could inhibit abnormally high expressions of TGFBR2 induced by radiation,and had significant protective effect on cell apoptosis and G2/M phase arrest induced by radiation.(3)The protective effect of TGF-β3 on radiation-induced apoptosis and G2/M cell cycle arrest was lost after TGFBR2 interference,suggesting that TGF-β3 exerted its inhibitory effect on radiation-induced lung epithelial cell injury through TGFBR2.Conclusion TGF-β3 can protect radiation-damaged lung epithelial cells through TGFBR2.

关 键 词：转化生长因子-Β3 Ⅱ型转化生长因子-β受体 电离辐射 肺上皮细胞损伤 肌动蛋白类 细胞凋亡 细胞周期 

分 类 号：R818.74[医药卫生—放射医学]