沙利度胺抑制TGF-β_1诱导的HELF细胞中结缔组织生长因子基因启动子的激活  

Thalidomide inhibits TGF-β_1-induced activation of CTGF gene promoter in HELF cell line

作　　者：王达安[1,2] 林逸心 王峥[3] 桑朝磊 陆大祥[1,2] 王华东[1,2] 胡巢凤[1,2] 魏伟[1,2] 蒋建伟[4] 付咏梅[1,2] 李红梅[1,2] 

机构地区：[1]暨南大学医学院病理生理学系,广东广州510632 [2]暨南大学医学院国家中医药管理局病理生理学重点实验室,广东广州510632 [3]暨南大学医学院口腔系2010级学生,广东广州510632 [4]暨南大学医学院生物化学系,广东广州510632

出　　处：《中国病理生理杂志》2014年第4期693-697,共5页Chinese Journal of Pathophysiology

基　　金：广东省医学科研基金(No.A2010317;No.A2012317);暨南大学国家中医药管理局病理生理学重点实验室开放基金(No.2011ZD002);广东省大学生创新创业训练计划(No.1055912179)

摘　　要：目的:探讨沙利度胺(thalidomide,THD)对转化生长因子β1(transforming growth factorβ1,TGF-β1)诱导的人胚肺成纤维细胞(human embryonic lung fibroblast,HELF)结缔组织生长因子(connective tissue growth factor,CTGF)基因启动子激活的影响。方法:构建含有人类CTGF基因启动子的报告基因载体pGL3-CTGFP,将其瞬时转染HELF细胞,通过检测萤光素酶的活性,观察TGF-β1和THD对CTGF基因启动子活性的影响。结果:TGF-β1能以剂量依赖方式和时间依赖方式显著增加HELF细胞中报告基因的活性(P<0.05),最佳剌激浓度是5μg/L,萤光素酶相对活性为对照组的2.16倍,最佳剌激时间为12 h,萤光素酶相对活性为对照组的2.52倍;THD对报告基因的基础活性无明显影响(P>0.05),但以剂量依赖方式显著抑制TGF-β1上调报告基因活性的效应(P<0.05)。结论:TGF-β1能以剂量依赖方式和时间依赖方式上调HELF细胞中CTGF基因启动子的活性,而THD能以剂量依赖方式抑制此过程。AIM ： To investigate the effects of thalidomide （THD） on the activation of connective tissue growth factor （CTGF） gene promoter induced by transforming growth factor B1 （TGF-B1） in human embryonic lung fibroblasts （HELF）. METHODS： DNA sequence of CTGF gene promoter was cloned into luciferase reporter gene vector to construct the recombinant eukaryotic expression vector pGI.3-CTGFP, and the recombinant vector was transfected into HELF cell line. The effects of TGF-B1 and THD on the activation of CTGF gene promoter were detected by dual-luciferase analysis. RESULTS ： TGF-131 increased the reporter gene activity dose-dependently （ P 〈 0.05 ）, with a plateau at 5 ug/L being 2.16 folds as high as the control. TGF-B1-induced increase in the reporter gene activity was also time-dependent （ P 〈 0. 05）. After exposure to TGF-B1 （5 ug/L）, the level of luciferase activity reached its peak at 12 h and was 2.52 folds as high as the control. THD significantly inhibited TGF-13t-induced increase in the reporter gene activity in a dose-dependent manner, but its basal activity was not changed. CONCLUSION： TGF-B1 stimulates the transcriptional activity of CTGF gene promoter in HELF cells in a dose- and time-dependent manner, while THD may inhibit the effects dose-dependently.

关 键 词：沙利度胺 人胚肺成纤维细胞 结缔组织生长因子 

分 类 号：R363[医药卫生—病理学]