TAZ蛋白在血管紧张素Ⅱ诱导的人脐静脉内皮细胞MCP-1表达中的作用研究  

The Role of TAZ in AngiotensinⅡ-Induced MCP1 Expression in Human Umbilical Vein Endothelial Cells

作　　者：卓坤萍 姚阳[1] 周明生 徐茜 ZHUO Kunping;YAO Yang;ZHOU Mingsheng;XU Qian(College of Basic Medicine,Shenyang Medical College,Shenyang Liaoning 110034,China)

机构地区：[1]沈阳医学院基础医学院,辽宁沈阳110034

出　　处：《中国继续医学教育》2022年第17期190-194,共5页China Continuing Medical Education

基　　金：国家自然科学基金资助项目(81970357);辽宁省科学技术计划项目(2019-ZD-0337)。

摘　　要：目的观察PDZ结合域的转录共刺激因子(transcriptional co-activator with PDZ-binding motif,TAZ)蛋白对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)单核趋化因子1(monocyte chemoattractant protein-1,MCP-1)表达的影响。方法2020年3月—2021年4月,本研究首先给予TAZ抑制剂维替泊芬(verteporfin,Ve)预处理细胞1 h后,加入AngⅡ(0.1μM),将细胞分为(1)对照组:培养液;(2)AngⅡ组;(3)AngⅡ+Ve组;(4)Ve组,Ve(0.5μM)。培养24 h进行后续实验。其次给予蛋白磷酸酶2A(protein phosphatase 2A,PP2A)抑制剂LB-100(0.1μM)预处理细胞1 h后,再加入AngⅡ(0.1μM),将细胞分为(1)对照组:培养液;(2)AngⅡ组;(3)AngⅡ+LB-100组;(4)LB-100组。培养24 h后进行后续实验。使用免疫荧光法检测AngⅡ诱导的人脐静脉内皮细胞TAZ相对荧光强度,Western blot检测MCP-1表达,给予PP2A抑制剂LB-100预处理后,加入AngⅡ,测量各组TAZ、p-TAZ、p-PP2A/PP2A及MCP-1蛋白表达水平。结果AngⅡ增加人脐静脉内皮细胞TAZ相对荧光强度并上调MCP-1蛋白,抑制TAZ减少MCP-1表达。抑制PP2A升高AngⅡ诱导的p-PP2A/PP2A、p-TAZ表达、降低TAZ和MCP-1蛋白表达。结论AngⅡ可能通过激活PP2A-TAZ通路促进HUVEC细胞MCP-1的表达。Objective To investigated the role of TAZ in AngⅡ-induced expression of MCP-1 in HUVECs.Methods From March 2020 to April 2021,cells was pretreated with verteporfin(Ve),a TAZ inhibitor for 1 h,and AngⅡ(0.1μm)was added cultured for 24 h.Cells were divided into four groups:Control(Ctr)group,AngⅡgroup,AngⅡ+Ve group,Ve group(0.5μM).Then cells was pretreated with LB-100,a PP2A inhibitor for 1 h,and AngⅡ(0.1μm)was added cultured for 24 h.Cells were divided into four groups:Ctr group,AngⅡgroup,AngⅡ+LB-100 group,LB-100 group(0.1μM).The relative fluorescence intensity of TAZ was detected by immunofluorescence,and the protein expressions of MCP-1 was measured by Western blot.After pretreatment with PP2A inhibitor LB-100,AngⅡwas added to measure the protein expression of TAZ,p-TAZ,PP2A,p-PP2A and MCP-1 in each group.Results AngⅡincreased the relative fluorescence intensity of TAZ,the MCP-1 expression.Inhibiton of TAZ reduced MCP-1 expression.AngⅡinducded the expressions of p-PP2A/PP2A,p-TAZ increased and TAZ,MCP-1 decreased through treatment with PP2A inhibitor LB-100.Conclusion AngⅡmay promote the MCP-1 expression through activation of the PP2A-TAZ pathway.

关 键 词：血管紧张素Ⅱ 人脐静脉内皮细胞 PP2A TAZ 单核趋化因子1 炎症 

分 类 号：R392[医药卫生—免疫学]