摘要
目的:探讨lncRNA FUT8-AS1调控miR-139-5p对糖尿病(DM)合并脑梗死(CI)大鼠血管的影响。方法:将雄性Wistar大鼠随机分为对照组、DM+CI组、Vector组、shFUT8-AS1组、miR-139-5p Agomir组、pcDNA-FUT8-AS1组和pcDNA-FUT8-AS1+miR-139-5p Agomir组。采用TTC染色检测CI面积,免疫组化染色检测血管内皮生长因子(VEGF)和FMS样酪氨酸激酶(FLT-1)蛋白表达,并检测内皮依赖性微血管密度(MVD)。实时荧光定量PCR(RT-qPCR)法检测FUT8-AS1、VEGF、FLT-1 mRNA表达。Western blotting检测VEGF蛋白表达。双荧光素酶实验检测FUT8-AS1与miR-139-5p的靶向关系。结果:与对照组比较,DM+CI组CI面积显著增加,脑组织FUT8-AS1、FLT-1 mRNA表达上调,VEGF mRNA及蛋白表达上调,miR-139-5p表达下调(均P<0.05)。与Vector组相比,pcDNA-FUT8-AS1组FLT-1和VEGF mRNA及蛋白表达上调,MVD增加(均P<0.05);shFUT8-AS1组和miR-139-5p Agomir组FLT-1和VEGF mRNA及蛋白表达下调,MVD减少(均P<0.05)。与pcDNA-FUT8-AS1组相比,pcDNA-FUT8-AS1+miR-139-5p Agomir组FLT-1和VEGF mRNA及蛋白表达下调,MVD减少(均P<0.05)。结论:过表达FUT8-AS1通过靶向miR-139-5p上调VEGF和FLT-1表达,促进DM合并CI大鼠血管生成。
Objective:To investigate the effect of lncRNA FUT8-AS1 regulating miR-139-5p on angiogenesis in diabetes mellitus(DM)rats complicated with cerebral infarction(CI).Methods:The male Wistar rats were ran-domly divided into control group,DM+CI group,DM+CI group,Vector group,shFUT8-AS1 group,miR-139-5p Agomir group,pcDNA-FUT8-AS1 group and pcDNA-FUT8-AS1+miR-139-5p Agomir group.The area of CI was detected by TTC staining.The protein expression of vascular endothelial growth factor(VEGF)and FMS-like tyrosine kinase(FLT-1)was detected by immunohistochemistry and the endothelium-dependent microvessel density(MVD)was detected.The mRNA expression of FUT8-AS1,VEGF and FLT-1 was detected by real-time fluorescence quantitative PCR(RT-qPCR),the expression of VEGF protein was detected by western blotting and the targeting relationship between FUT8-AS1 and miR-139-5p was detected by dual luciferase assay.Results:Compared with the control group,the area of CI was significantly increased,the expression of FUT8-AS1,FLT-1 mRNA,and VEGF mRNA and protein was up-regulated,and the expression of miR-139-5p was down-regulated in the DM+CI group(all P<0.05).Compared with the Vector group,the expression of FLT-1 and VEGF mRNA and protein was up-regulated,and MVD was increased in the pcDNA-FUT8-AS1 group(all P<0.05);the expres-sion of FLT-1 and VEGF mRNA and protein was down-regulated in the shFUT8-AS1 group and the miR-139-5p Agomir group,and MVD was decreased(all P<0.05).Compared with the pcDNA-FUT8-AS1 group,the expres-sion of FLT-1 and VEGF mRNA and protein was down-regulated,and MVD was decreased in the pcD-NA-FUT8-AS1+miR-139-5p Agomir group(all P<0.05).Conclusion:Overexpression of FUT8-AS1 up-regulates the expression of VEGF and FLT-1 by targeting miR-139-5p to promote angiogenesis in DM rats complicated with CI.
作者
谷骞
柳晓锋
Gu Qian;Liu Xiaofeng(Department of Neurology,Shijiazhuang People’s Hospital,Shijiazhuang 050000,China)
出处
《广西医科大学学报》
CAS
2023年第12期2017-2022,共6页
Journal of Guangxi Medical University
基金
河北省2023年度医学科学研究课题计划资助项目(No.20231620)。
