Takeda G蛋白偶联受体5在血管平滑肌细胞增殖和迁移中的作用  

Role of TGR5 in proliferation and migration of vascular smooth muscle cells

作　　者：杨曦 张黎 杨耀 王嘉 孙雄山[2] 王强 YANG Xi;ZHANG Li;YANG Yao;WANG Jia;SUN Xiong-shan;WANG Qiang(Dept of Clinical College of Medicine,Southeast Medicine University,Luzhou,Sichuan 646000,China;Dept of Cardiology,the General Hospital of Western Theater Command,Chengdu 610083,China;Dept of Pharmacy,the General Hospital of Western Theater Command,Chengdu 610083,China;College of Medicine,Southwest Jiaotong University,Chengdu 610031,China)

机构地区：[1]西南医科大学临床医学院心血管内科,四川泸州646000 [2]西部战区总医院心血管内科,四川成都610083 [3]西部战区总医院药剂科,四川成都610083 [4]西南交通大学医学院,四川成都610031

出　　处：《中国药理学通报》2024年第8期1447-1454,共8页Chinese Pharmacological Bulletin

基　　金：四川省自然科学基金资助项目(No.2022NSFSC0820);国家自然科学基金资助项目(No.82100419);西部战区总医院院管课题(No.2024-YGJC-A06,2021-XZYG-B28)。

摘　　要：目的探讨Takeda G蛋白偶联受体5(Takeda G protein-coupled receptor 5,TGR5)在小鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖和迁移中的作用及机制。方法用血小板衍生生长因子(platelet-derived growth factor,PDGF-BB)诱导VSMCs增殖、迁移,以INT-777特异性激活TGR5,CCK-8试剂盒及增殖细胞核抗原(Ki-67)免疫荧光染色用于检测细胞增殖能力,划痕试验用于检测细胞迁移能力。Western blot检测TGR5蛋白水平变化。为探究TGR5在血管内膜增生中的作用,将40只雄性野生型C57BL/6J小鼠随机分为假手术组、内膜损伤组、假手术+UDCA(熊去氧胆酸,TGR5激动剂)组及内膜损伤+UDCA组,每组10只。造模完成后按组分别予以口服普通维持饲料及含0.5%UDCA的普通维持饲料,持续21 d后分别取材,HE染色观察颈动脉内膜增生程度,Ki-67免疫荧光染色观察颈动脉内膜血管平滑肌增殖变化。结果特异性激活TGR5明显降低VSMCs增殖活力及Ki-67阳性细胞率,同时使VSMCs划痕愈合速度减慢。特异性激活TGR5使细胞内UCP2表达增加、活性氧(reactive oxygen species,ROS)水平降低。过氧化氢恢复细胞内ROS水平后,TGR5抑制VSMCs增殖迁移的作用被削弱。激活TGR5能减轻颈动脉损伤后内膜增生。结论TGR5可能通过UCP2改善细胞内氧化应激,从而抑制小鼠VSMCs的增殖和迁移。Aim To explore the role of Takeda G protein-coupled receptor 5(TGR5)in the proliferation and migration of vascular smooth muscle cells(VSMCs)within the intimal layer of mice.Methods Mouse VSMCs were stimulated for proliferation and migration with PDGF-BB,followed by administration of INT-777 for activation of TGR5.CCK-8 assay and Ki-67 immunofluorescence staining were employed to evaluate cell proliferation.The scratch assay was utilized to assess migration.Western blot analysis was conducted to monitor TGR5 protein expression.To further investigate the role of TGR5 in intimal hyperplasia in vivo,20 male wild-type C57BL/6J mice were randomly divided into four groups:sham group,carotid artery endothelial injury group,sham+UDCA(ursodeoxycholic acid,a TGR5 agonist)group,and carotid artery endothelial injury+UDCA group(n=5 in each group).After the establishment of endothelial injury model,mice were orally fed with regular maintenance feed containing 0.5%UDCA for 21 days.Subsequently,samples were collected for Hematoxylin and Eosin(HE)staining to assess the neointimal hyperplasia.Immunofluorescence(IF)staining was used to examine the proliferation of VSMCs within the carotid intima.Results The specific activation of TGR5 marked-ly diminished cell viability,the proportion of Ki-67-positive cells,and slowed down the rate of wound-healing.Notably,the specific activation of TGR5 increases the expression of UCP2 in cells and reduces the levels of reactive oxygen species(ROS).The inhibitory effect of TGR5 on VSMC proliferation and migration was neutralized upon the restoration of intracellular ROS level with H 2O 2.Activation of TGR5 was found to mitigate intimal thickening following carotid artery injury.Conclusion TGR5 may inhibit the proliferation and migration of mouse VSMCs by attenuating intracellular oxidative stress.

关 键 词：细胞增殖 细胞迁移 血管平滑肌 再狭窄 G蛋白偶联胆汁酸受体1 INT-777 

分 类 号：R-332[医药卫生] R322.12R322.74R329.28R543R977.6