RASAL1在同型半胱氨酸致肺血管内皮通透性增加中的作用及机制研究  

Role and mechanism of RASAL1 in increase of pulmonary vascular endothelial permeability induced by homocysteine

作　　者：李昕怡 秦凯悦 杨梓尧 张玲[4] 李珍 杨亚丽 徐茜 杨晓玲 LI Xin-yi;QIN Kai-yue;YANG Zi-yao;ZHANG Ling;LI Zhen;YANG Ya-li;XU Xi;YANG Xiao-ling(Key Laboratory of Metabolic Cardiovascular Disease Research,National Health Commission;Dept of Pathology,Ningxia People's Hospital;The Second Clinical Medical Collage of Ningxia Medical University;Dept of Pathology,General Hospital of Ningxia Medical University;School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]国家卫健委代谢性心血管疾病研究重点实验室 [2]宁夏医科大学基础医学院 [3]宁夏医科大学第二临床医学院 [4]宁夏人民医院病理科 [5]宁夏医科大学总医院病理科,宁夏银川750004

出　　处：《中国药理学通报》2024年第8期1454-1461,共8页Chinese Pharmacological Bulletin

基　　金：国家自然基金资助项目(No.81960063);宁夏自然基金资助项目(No.2021AAC2012)。

摘　　要：目的探讨同型半胱氨酸(homocystein,Hcy)对肺微血管内皮细胞(pulmonary microvascular endothelial cells,PMVECs)通透性的影响及Ras蛋白样激活剂(Ras protein activator like,RASAL1)的作用及机制。方法CBS^(+/-)小鼠喂养高蛋氨酸饮食(HMD)16周复制高同型半胱氨酸血症(HHcy)动物模型;HE染色观察肺组织结构变化;qRT-PCR检测肺组织RASAL1、DNMT1 mRNA水平;Western blot检测RASAL1、DNMT1、ZO-1、VE-cadherin蛋白表达;MS-PCR检测RASAL1启动子区甲基化;PMVECs转染Ad-RASAL1检测ZO-1和VE-cadherin表达;si-DNMT1干扰片段转染PMVECs,检测RASAL1表达。结果HMD小鼠血清Hcy水平明显增加,HE染色显示肺组织结构严重紊乱,RASAL1、ZO-1、VE-cadherin表达降低而DNMT1表达增加,RASAL1启动子区甲基化程度增加。PMVECs转染Ad-RASAL1后ZO-1和VE-cadherin表达增加;敲低DNMT1后,RASAL1表达增加。结论Hcy可以导致PMVECs通透性增加,其机制与上调RASAL1甲基化水平有关。Aim To investigate the effect of homocysteine(Hcy)on the permeability of pulmonary microvascular endothelial cells(PMVECs)and the role and mechanism of RASAL1.Methods CBS^(+/-)mice were fed a high methionine diet(HMD)for 16 weeks to replicate an animal model of hyperhomocysteinemia(HHcy).HE staining was used to observe the changes in lung tissue structure.qRT-PCR was used to detect the levels of RASAL1 and DNMT1 mRNA in lung tissue.Western blot was used to detect the expression of RASAL1,DNMT1,ZO-1,and VE cadherin proteins.Methylation specific PCR was used to detect methylation in the RASAL1 promoter region.PMVECs were transfected with Ad-RASAL1 to detect the expression of ZO-1 and VE cadherin.The si-DNMT1 interference fragment was transfected into PMVECs,and the expression of the RASAL1 was detected by qRT-PCR and Western blot.Results Serum Hcy level of HMD mice was significantly raised,and HE staining showed severe structural disorder in lung tissue.The expression of RASAL1,ZO-1,and VE cadherin was decreased,while the expression of DNMT1 was increased.The degree of methylation in the RASAL1 promoter region was raised.The expression of ZO-1 and VE cadherin increased after PMVECs were transfected with Ad-RASAL1.After knocking down DNMT1,RASAL1 expression was increased.Conclusion Hcy can increase the permeability of PMVECs,and its mechanism is related to the upregulation of RASAL1 methylation level.

关 键 词：同型半胱氨酸 RASAL1 肺微血管内皮细胞 通透性 DNA甲基化 DNMT1 

分 类 号：R-332[医药卫生] R322.35R341.7R394.2R977.4