TL1A在高糖诱导的人脐静脉内皮细胞凋亡中的作用及机制探讨

目的 研究肿瘤坏死因子配体相关分子1A（TL1A）在高糖诱导的人脐静脉内皮细胞凋亡中的作用,并探讨机制.方法 采用RT-PCR、Western blot法检测不同浓度（5.6、11.2、22.4、33.6 mmol/L）葡萄糖与人脐静脉内皮细胞共同孵育48 h以及22.4 mmol/L葡萄糖作用于内皮细胞12、24、48、96 h后,TL1A mRNA及蛋白质在人脐静脉内皮细胞中的表达情况.在22.4 mmol/L葡萄糖培养的人脐静脉内皮细胞中加入不同浓度的TL1A单克隆抗体（1、3、9 μg/mL）,48 h后进行Annexin V/PI双染色流式细胞凋亡计数,以纯化重组人TL1A作为内源性TL1A的对照.结果 在人脐静脉内皮细胞中,随着葡萄糖浓度升高及作用时间延长,TL1A mRNA和蛋白水增高（P＜0.05）.流式细胞计数显示,高糖培养人脐静脉内皮细胞经TL1A单克降抗体处理后凋亡明显减少（P＜0.05）.结论 高浓度葡萄糖诱导体外培养的人脐静脉内皮细胞TL1A的表达呈浓度、时间依赖性增高;TL1A表达增多可能是高糖诱导人脐静脉内皮细胞凋亡的重要因素.

Effects of TL1A on the high glucose induced apoptosis in human vascular endothelial eells and its mechanism

Objective To investigate the effects and mechanism of TNF-like ligand 1 aberrance （TL1A） on the high glucose induced apoptosis in human umbilical vein endothelial cells （HUVECs）.Methods HUVECs were treated with different concentrations of glucose （5.6,11.2,22.4,33.6 mmol/L） for 48 hours or 22.4 mmol/L glucose for different time （12,24,48,96 h） in vitro.The expression of TL1A was evaluated by RT-PCR and Western blot.Adding different concentrations of TL1A monoclonal antibody （1,3,9 μg/mL） to HUVECs which were cultured in 22.4 mmol/L glucose,taking recombinant human TL1 A as an endogenous control,then 48 hours later,annexin V/PI double staining flow cytometry was used to determine the HUVECs apoptosis rate.Results In high glucose conditions,the mRNA and protein expression of TL1 A in HUVECs were significantly higher than that in normal glucose group in a dose-and time-dependent manner （P ＜ 0.05）.The apoptosis rate of HUVECs was significantly decreased after treated with the monoclonal antibody of TL1 A （P ＜ 0.05）.Conclusion Expression of TL1A induced high glucose in cultured HUVECs increased in a dose-and timedependent manner,which played an important role in HUVECs apoptosis induced by high glucose.