转化生长因子-β1促血管内皮细胞向间质细胞转化的作用研究  

Role of TGF-β1 in promoting transformation of intravascular endothelial cells into interstitial cells

作　　者：简宇[1] 简华刚[1] 黄宏[2] 徐祥[2] 郭敏[1] 代卉[2] 崔文慧[2] 蒋建新[3] 朱佩芳[3] 王正国[3] 

机构地区：[1]重庆医科大学附属第二医院创伤烧伤科,重庆400016 [2]第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室第一研究室,重庆400042 [3]第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室第四研究室,重庆400042

出　　处：《第三军医大学学报》2010年第21期2307-2310,共4页Journal of Third Military Medical University

基　　金：国家重点基础研究发展计划(973计划)(2005CB522603);国家自然科学基金(30871025)~~

摘　　要：目的观察转化生长因子-β1(transforming growth factor-β1,TGF-β1)促进内皮细胞向间质细胞转化[endothelial-mesenchymal(myofibroblast)transition,EnMT]的效应,探讨肌成纤维细胞的来源以及纤维化疾病发生的新机制。方法分离培养人脐静脉内皮细胞,采用不同浓度的TGF-β1(0、10、25、和50ng/ml)刺激细胞72h后,倒置显微镜观察诱导后细胞形态变化,免疫荧光方法检测第Ⅷ因子相关抗原和α-SMA表达变化,RT-PCR检测VE-钙粘蛋白(VE-Cadherin),α-SMA和Ⅰ型胶原基因表达变化。结果正常对照组内皮细胞呈铺路石样生长,TGF-β1刺激组内皮细胞由铺路石样形态向梭形转化。免疫荧光检测可见对照组含有少量FⅧ,α-SMA双阳性细胞。TGF-β1组FⅧ,α-SMA双阳性细胞明显增多,并且具有明显的浓度依赖性(P<0.05,P<0.01)。RT-PCR检测结果显示,TGF-β1刺激导致内皮细胞特异性标记物VE-钙粘蛋白(VE-cadherin)基因的表达显著减少(P<0.05),随着TGF-β1刺激浓度的增加,VE-钙粘蛋白基因表达呈明显的下降趋势;相反,内皮细胞间质性标记物α-SMA和I型胶原基因的表达显著增加(P<0.05,P<0.01),并且具有明显的浓度依赖性。结论 TGF-β1刺激内皮细胞后,细胞表型和功能均表现为间质细胞特性,提示TGF-β1具有促进内皮细胞向间质细胞转化(EnMT)的作用,并且其促EnMT效应具有明显的浓度依赖性。Objective To observe the role of TGF-β1 in promoting transformation of endothelial cells into interstitial cells and study the source of myofibroblasts and the mechanism underlying fibrosis.Methods Endothelial cells were isolated from human umbilical vein and cultured with different concentrations of TGF-β1（0,10,25 and 50 ng/ml）for 72 h.Then,changes of cell morphology were observed under an inverted phase contrast microscope.Expression of factor Ⅷ related antigen and α-Smooth muscle actin（α-SMA）was detected by immunofluorescence assay.Expression of VE-Cadherin,α-SMA and type 1 collagen was detected by RT-PCR.Results A paving stone-like growth and a spindle-like growth of endothelial cells was observed in control group and TGF-β1 stimulating group,respectively.Immunofluorescence assay showed a small number of factor Ⅷ and α-SMA positive cells in control group.The number of factor Ⅷ and α-SMA positive cells was significantly greater in TGF-β1 group than in control group（P0.05）and increased with the concentration of TGF-β1.RT-PCR showed that the expression level of VE-Cadherin significantly decreased with the concentration of TGF-β1（P0.05）while the expression level of α-SMA and type 1 collagen significantly increased with concentration of TGF-β1（P0.05）.Conclusion TGF-β1 promotes the transformation of endothelial cells into interstitial cells in a concentration-dependent manner.

关 键 词：内皮细胞 肌成纤维细胞 转分化 转化生长因子-Β1 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]