miR-30b-5p通过Sema3A调控HUVECs细胞增殖、凋亡、迁移与血管形成  

miR-30b-5p Regulates Proliferation,Apoptosis,Migration and Angiogenesis in HUVECs through Sema3A

作　　者：李志杰 LI Zhi-jie(Department of Endocrinology,Sinopharm Tongmei General Hospital,Datong 037009,China)

机构地区：[1]国药同煤总医院内分泌科,山西大同037009

出　　处：《南昌大学学报（医学版）》2023年第5期28-33,共6页Journal of Nanchang University：Medical Sciences

基　　金：山西省卫计委科研项目(20141008)。

摘　　要：目的探究miR-30b-5p对HUVECs细胞增殖、凋亡、迁移与血管形成的影响及机制。方法培养的HUVECs细胞进行不同处理:blank组(正常糖培养)、HG组(高糖培养)、HG+mimic-NC组(高糖联合mimic-NC转染)、HG+miR-30b-5p mimic组(高糖联合miR-30b-5p mimic转染)、HG+miR-30b-5p mimic+oe-NC组(高糖联合miR-30b-5p mimic+oe-NC转染)、HG+miR-30b-5p mimic+oe-Sema3A组(高糖联合miR-30b-5p mimic+oe-Sema3A转染)。CCK-8、流式细胞术、划痕实验、血管生成实验分别检测细胞增殖、凋亡、迁移和血管形成。qRT-PCR检测miR-30b-5p、Sema3A mRNA表达。Starbase、Targetscan、miRDB网站预测miR-30b-5p与Sema3A的靶向结合位点,双荧光素酶报告实验分析miR-30b-5p对Sema3A的靶向调控。结果与blank组比较,HG组HUVECs细胞miR-30b-5p表达下降,细胞增殖、迁移和血管形成能力降低,凋亡率增加(均P<0.05);与HG+mimic-NC组比较,HG+miR-30b-5p mimic组细胞的增殖、迁移和血管形成能力增强,凋亡率降低(均P<0.05);miR-30b-5p靶向调控Sema3A;与HG+miR-30b-5p mimic+oe-NC组比较,HG+miR-30b-5p mimic+oe-Sema3A组细胞增殖、迁移和血管形成能力降低,凋亡率增加(均P<0.05)。结论miR-30b-5p通过负调控Sema3A促进高糖介导的HUVECs细胞增殖、迁移和血管形成,抑制其细胞凋亡。Objective To explore the effect of miR-30b-5p on the proliferation,apoptosis,migration and angiogenesis in human umbilical vein endothelial cells(HUVECs)and its mechanisms.Methods HUVECs were treated with normal glucose(blank group),high glucose(HG group),high glucose combined with mimic-NC transfection(HG+mimic-NC group),high glucose combined with miR-30b-5p mimic transfection(HG+miR-30b-5p mimic group),high glucose combined with miR-30b-5p mimic+oe-NC transfection(HG+miR-30b-5p mimic+oe-NC group),or high glucose combined with miR-30b-5p mimic+oe-Sema3A transfection(HG+miR-30b-5p mimic+oe-Sema3A group).Cell proliferation,apoptosis,migration and angiogenesis were detected by CCK-8 assay,flow cytometry,scratch test and tube formation assay,respectively.The expression of miR-30b-5p and Sema3A mRNA was detected by qRT-PCR.The Starbase,Targetscan and miRDB were employed to predict the targeted binding sites of miR-30b-5p and Sema3A.Dual luciferase report assay was performed to analyze the targeted regulation of miR-30b-5p on Sema3A.Results Compared with the blank group,the expression of miR-30b-5p and the proliferation,cell migration and angiogenesis of cells were reduced,and the apoptosis rate was increased in HG group(P<0.05).Compared with HG+mimic-NC group,the proliferation,migration and angiogenesis of cells were increased,and the apoptosis rate was decreased in HG+miR-30b-5p mimic group(P<0.05).miR-30b-5p exerted a targeted regulatory effect on Sema3A.Compared with HG+miR-30b-5p mimic+oe-NC group,the proliferation,migration and angiogenesis of cells were reduced,and the apoptosis rate was elevated in HG+miR-30b-5p mimic+oe-Sema3A group(P<0.05).Conclusion miR-30b-5p promotes high glucose-mediated proliferation,migration and angiogenesis and suppresses apoptosis in HUVECs through negatively regulating Sema3A.

关 键 词：糖尿病足病 miR-30b-5p 臂板蛋白3A多肽 靶向调控 血管生成 

分 类 号：R587.2[医药卫生—内分泌]