Exendin-4通过调控SIRT1/P53信号通路改善高糖作用下的内皮祖细胞功能障碍  

Exendin-4 attenuates high glucose-induced endothelial progenitor cell dysfunction by regulating SIRT1/p53 signaling pathway

作　　者：赵祺 涂强 谢华强[1] 曹政[1,2] ZHAO Qi;TU Qiang;XIE Hua-qiang;CAO Zheng(Dept of Cardiology, Taihe Hospital, Shiyan Hubei 442000, China;Hubei University of Medicine, Shiyan Hubei 442000, China)

机构地区：[1]十堰市太和医院,湖北十堰442000 [2]湖北医药学院,湖北十堰442000

出　　处：《中国药理学通报》2022年第4期538-544,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 81771522);湖北省卫生健康委员会项目(No WJ2021M258)。

摘　　要：目的研究Exendin-4对高糖诱导的内皮祖细胞(endothelial progenitor cells,EPCs)功能障碍的影响,并探讨沉默信息调节因子1(silent information regulator 1,SIRT1)蛋白在其中的作用。方法采用密度梯度离心法分离并培养健康志愿者外周血EPCs,将不同浓度Exendin-4(12.5、50、100、200μmol·L-1)联合高糖(25 mmol·L-1)处理EPCs并检测其活力,选择药物的最佳作用浓度。将最佳浓度的Exendin-4与高糖共处理EPCs 72 h后检测其功能活性。通过迁移、黏附、小管形成实验检测EPCs功能;同时对乳酸脱氢酶(LDH)和丙二醛(MDA)水平进行检测。使用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测TNF-α、IL-1β及IL-6 mRNA的表达。使用Western blot来检测SIRT1、p53和Ac-p53的蛋白表达。结果Exendin-4处理可明显改善高糖作用下EPCs的活力,尤以50μmol·L-1时作用效果最佳(P<0.05)。与高糖组相比,Exendin-4处理可上调SIRT1蛋白表达(P<0.05),增加高糖作用下EPCs的体外迁移、黏附、小管形成能力(P<0.05),并改善LDH及MDA水平(P<0.05),同时抑制TNF-α、IL-β及IL-6的mRNA表达(P<0.05)。而SIRT1受体抑制剂(EX-527)可使Exendin-4改善EPCs功能活性的作用受到明显的抑制(P<0.05)。另外,使用SIRT1激动剂(SRT1720)亦可改善高糖诱导的EPCs功能障碍(P<0.05)。结论Exendin-4可提高高糖作用下EPCs活力,改善EPCs功能,抑制氧化应激损伤,降低炎症因子表达,其机制可能与调控SIRT1/p53信号通路有关。Aim To investigate the effect of Exendin-4 on endothelial progenitor cells(EPCs)induced by high glucose and the function of silent information regulator 1(SIRT1).Methods EPCs were isolated and cultured by density gradient centrifugation from peripheral blood of healthy volunteers.Different concentrations of Exendin-4(12.5,50,100,200μmol·L-1)induced EPCs were respectively performed by activity test to select the appropriate concentration.The optimal concentrations of Exendin-4 and high glucose(25 mmol·L-1)were incubated together for 72 h to detect the functional activities of EPCs.The capabilities of migration,adhension and tube formation of EPCs in vitro were detected respectively.The levels of LDH and MDA in EPCs were detected.The mRNA expressions of TNF-α,IL-1βand IL-6 in EPCs were measured by real-time fluorescent quantitative PCR(RT-qPCR).The protein expression of SIRT1,P53 and Ac-P53 in EPCs were determined by Western blot.Results Exendin-4 could increase the viability of EPCs induced by high glucose in a dose-dependent manner,especially for 50μmol·L-1(P<0.05).The results of Western blot showed that the protein expression of SIRT1 was significantly enhanced by 50μmol·L-1 Exendin-4 treatment(P<0.05).Compared with high glucose group,Exendin-4 significantly increased the migration,adhesion,tube formation of EPCs(P<0.05)and decreased the level of LDH and MDA(P<0.05).The mRNA expression of TNF-α,IL-βand IL-6 in EPCs also decreased(P<0.05).However,the protective effects of Exendin-4 could be significantly blocked by SIRT1 inhibitor(EX-527)(P<0.05).In addition,the SIRT1 agonist(SRT1720)could also improve the dysfunction of EPCs induced by high glucose(P<0.05).Conclusions Exendin-4 can improve the viability of human EPCs,restore the EPCs normal function,reduce high glucose-induced oxidative damage,and reduce the releases of inflammatory cytokines under high glucose condition,which may be related to the regulation of SIRT1/P53 signaling pathway.

关 键 词：EXENDIN-4 胰高血糖素样肽-1 内皮祖细胞 糖尿病 血管生成 沉默信息调节子1 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R341.6[医药卫生—基础医学]