内皮祖细胞损伤来源微粒对内皮祖细胞的影响机制研究  

Study on the mechanism of the effect of endothelial progenitor cell injury-derived microparticles on endothelial progenitor cells

作　　者：马艺萍 阿不都热合曼·米吉提 袁玉娟 吐尔孙阿依·依斯米提拉 阿卜拉江·艾合麦提 穆叶赛·尼加提[3] MA Yiping;ABUDUREHEMAN Mijiti;YUAN Yujuan;TUERSUNAYI Yisimitila;ABULAJIANG Aihemati;MUYESAI Nijiati(Graduate School,Xinjiang Medical University,Urumqi,Xinjiang 830000,China;Department of Cardiovascular,the Second People’s Hospital of Kashgar,Kashgar,Xinjiang 844099,China;Emergency Center,Xinjiang Uygur Autonomous Region People’s Hospital,Urumqi,Xinjiang 830000,China)

机构地区：[1]新疆医科大学研究生院,乌鲁木齐830000 [2]喀什地区第二人民医院心血管内科,新疆喀什844099 [3]新疆维吾尔自治区人民医院急救中心,乌鲁木齐830000

出　　处：《重庆医学》2023年第23期3538-3545,共8页Chongqing medicine

基　　金：国家自然科学基金地区基金项目(82060076);新疆维吾尔自治区研究生创新项目(XJ2023G202)。

摘　　要：目的研究不同损伤处理的内皮祖细胞(EPC)来源微粒(MPs)对EPC的影响。方法培养EPC并使用流式细胞术鉴定,用高糖(HG)、肿瘤坏死因子(TNF)-α重组蛋白处理EPC,进而提取MPs。用透射电镜观察EPC内微管、高尔基体等细胞器的结构变化,MTT检测细胞增殖情况,细胞划痕实验评价细胞迁移能力,细胞管腔形成实验检测管腔形成情况。用实时荧光定量PCR(qPCR)和Western blot检测不同处理后EPC中内皮型一氧化氮合成酶(eNOS)、沉默信息调节因子1(SIRT1)、大鼠肉瘤(RAS)和细胞外调节蛋白激酶(ERK)的表达水平。结果与control-EPC-MPs组比较,HG-EPC-MPs组、TNF-α-EPC-MPs组微管结构完整,长度有所缩短,高尔基体结构相对完整。与control-EPC-MPs组比较,HG-EPC-MPs组、TNF-α-EPC-MPs组EPC增殖率下调,细胞迁移能力降低,成管减少(P<0.05)。与control-EPC-MPs组比较,HG-EPC-MPs组、TNF-α-EPC-MPs组eNOS、SIRT1、RAS和ERK的mRNA及其蛋白表达下调(P<0.05)。结论HG和TNF-α介导EPC损伤来源的MPs可能通过调控SIRT1/ERK1通路蛋白的表达,使EPC内含细胞器的结构改变,从而影响EPC的生物学功能。Objective To investigate the effects of endothelial progenitor cells(EPC)-derived microparticles(MPs)with different injury treatments on EPC.Methods EPC cells were cultured and EPCs were identified by flow cytometry.EPCs were treated with high glucose(HG)and tumor necrosis factor(TNF)-αrecombinant protein,and MPs were extracted.The structural changes of microtubules,Golgi bodies and other organelles in EPC were observed by transmission electron microscopy.MTT assay was used to detect cell proliferation.Cell scratch assay was used to evaluate cell migration ability.Cell lumen formation assay was used to detect lumen formation.The expression levels of endothelial nitric oxide synthase(eNOS),silent information regulator 1(SIRT1),rat sarcoma(RAS)and extracellular regulated protein kinase(ERK)in EPC after different treatments were detected by quantitative real-time fluorescence PCR(qPCR)and Western blot.Results Compared with the control-EPC-MPs group,the microtubule structure of the HG-EPC-MPs group and the TNF-α-EPC-MPs group was complete,the length was shortened,and the Golgi structure was relatively complete.Compared with the control-EPC-MPs group,the proliferation rate of EPC in the HG-EPC-MPs group and the TNF-α-EPC-MPs group was down-regulated,the cell migration ability was decreased,and the tube formation was decreased(P<0.05).Compared with the control-EPC-MPs group,the mRNA and protein expressions of eNOS,SIRT1,RAS and ERK in the HG-EPC-MPs group and the TNF-α-EPC-MPs group were down-regulated(P<0.05).Conclusion HG and TNF-αmediated MPs derived from EPC injury may change the structure of organelles in EPC by regulating the expression of SIRT1/ERK1 pathway proteins,thus affecting the biological function of EPC.

关 键 词：内皮祖细胞 微粒 SIRT1 ERK1 细胞损伤 机制研究 

分 类 号：R541[医药卫生—心血管疾病]