miR-106调控CC趋化因子配体2对增生型糖尿病视网膜病变中人视网膜微血管内皮细胞增殖、血管生成和炎症反应的影响  

miR-106 affects proliferation of human retinal microvascular endothelial cells,angiogenesis and inflammatory response in proliferative diabetic retinopathy via regulating CCL2

作　　者：田涛[1] 姚睿 彭婧利[1] 刘茹[1] 谢丽莲[1] 邝国平[1] 周小平[1] TIAN Tao;YAO Rui;PENG Jingli;LIU Ru;XIE Lilian;KUANG Guoping;ZHOU Xiaoping(Department of Ophthalmology,the First People’s Hospital of Chenzhou,Chenzhou 423000,Hunan Province,China;Department of Clinical Medicine,Xiangya School of Medicine,Central South University,Changsha 410006,Hunan Province,China)

机构地区：[1]郴州市第一人民医院眼科,湖南省郴州市423000 [2]中南大学湘雅医学院临床医学部,湖南省长沙市410006

出　　处：《眼科新进展》2021年第9期831-837,共7页Recent Advances in Ophthalmology

摘　　要：目的探讨miR-106调控CC趋化因子配体2(CCL2)对增生型糖尿病视网膜病变(PDR)中人视网膜微血管内皮细胞(HRMEC)增殖、血管生成、炎症反应的影响。方法GEO数据库筛选PDR中差异表达的miRNAs。高糖(25.0 mmol·L^(-1)葡萄糖,HG)诱导HRMEC建立PDR细胞模型,qRT-PCR检测miR-106和CCL2在正常葡萄糖(5.5 mmol·L^(-1)葡萄糖,NG)和HG培养条件下HRMEC中的表达。将PDR患者的血清外泌体做miR-106过表达处理后与HG处理的HRMEC共培养,同时干预HRMEC中CCL2的表达以探讨血清外泌体miR-106和CCL2在PDR中的功能。采用MTT法、小管形成实验和ELISA法分别检测各组HRMEC增殖、血管生成和炎症因子的表达。双荧光素酶报告实验用以验证miR-106和CCL2的靶向关系。结果与NG组细胞中miR-106表达(1.04±0.10)、CCL2表达(1.02±0.09)相比,HG培养的HRMEC中miR-106表达(0.68±0.06)降低,CCL2表达(1.38±0.11)升高(均为P<0.05)。PDR患者血清外泌体中miR-106表达较NDR患者血清外泌体中表达降低(P<0.05)。与HG+PDR-exo+miR-NC组相比,HG+PDR-exo+miR-106 mimic组HRMEC活力降低,血管生成被抑制,细胞上清液中TNF-α、IL-1β和IL-6水平降低,而SOD、CAT和GSH水平升高(均为P<0.05)。双荧光素酶报告实验证实了CCL2是miR-106的一个靶点。与HG+PDR-exo+miR-106 mimic+oe-NC组相比,HG+PDR-exo+miR-106 mimic+oe-CCL2组HRMEC活力提高,血管生成被诱导,TNF-α、IL-1β和IL-6水平升高,而SOD、CAT和GSH水平降低(均为P<0.05)。结论血清外泌体miR-106能够抑制CCL2表达,进而对PDR中的HRMEC增殖、血管生成和炎症反应起抑制作用。Objective To investigate the effects of miR-106 on the proliferation of human retinal microvascular endothelial cells(HRMECs),angiogenesis and inflammatory response in proliferative diabetic retinopathy(PDR)through regulating chemokine(CC-motif)ligand 2(CCL2).Methods The differentially expressed miRNAs in PDR was screened using the GEO database.HRMECs were induced by high glucose to establish the in vitro PDR model.Expression levels of miR-106 and CCL2 in HRMECs induced with normal glucose(NG)or high glucose(HG)were detected by qRT-PCR.Serum exosomes collected from PDR patients were transfected with miR-106 mimic,which were co-cultured with HG-treated HRMECs.Meanwhile,the expression of CCL2 in HRMECs was intervened to explore the functions of serum exosomal miR-106 and CCL2 in PDR.MTT,tube formation assay and ELISA were performed to detect the proliferation,angiogenesis and expression levels of inflammatory factors in HRMECs.Dual-luciferase reporter assay was used to verify the targeting relationship between miR-106 and CCL2.Results Compared with NG group,miR-106 expression was downregulated(1.04±0.10 vs.0.68±0.06),while CCL2 expression(1.02±0.09 vs.1.38±0.11)was upregulated in HG-induced HRMECs(both P<0.05).The expression level of miR-106 in serum exosomes of PDR patients was significantly lower than that in NDR patients(P<0.05).Overexpression of miR-106 in exosomes collected from PDR patients and co-culturing with HG-induced HRMECs resulted in reduced viability,inhibited angiogenesis,decreased levels of TNF-α,IL-1βand IL-6 in cell supernatant and increased activities of SOD,CAT and GSH(all P<0.05).Dual-luciferase reporter assay confirmed that CCL2 was the target of miR-106.Overexpression of CCL2 in HRMECs co-cultured with HG and exosomes collected from PDR patients that were overexpressed with miR-106 enhanced cell viability,inhibited angiogenesis,increased levels of TNF-α,IL-1βand IL-6 in cell supernatant and decreased activities of SOD,CAT and GSH than those without intervention of CCL2(all P<0.05).Co

关 键 词：血清外泌体 miR-106 CC趋化因子配体2 增生型糖尿病视网膜病变 

分 类 号：R774.1[医药卫生—眼科]