KIF11通过激活Wnt/β-catenin通路调控高糖诱导的人视网膜微血管内皮细胞损伤  

Regulation effect of kinesin family member 11 on the high glucose-induced injury of human retinal microvascular endothelial cells via activating Wnt/β-catenin pathway

作　　者：韩昌婧 徐丽[1] 段雪娟 艾欣 HAN Changjing;XU Li;DUAN Xuejuan;AI Xin(Department of Ophthalmology,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710103,Shaanxi Province,China;Department of Ophthalmology,Xi’an First Hospital,Institute of Ophthalmology,Key Lab of Ophthalmology,Clinical Center for Ophthalmology,Xi’an 710002,Shaanxi Province,China)

机构地区：[1]西安交通大学第二附属医院眼科,陕西省西安市710103 [2]西安市第一医院眼科,陕西省眼科研究所,陕西省眼科学重点实验室,陕西省眼科疾病临床医学研究中心,陕西省西安市710002

出　　处：《眼科新进展》2022年第12期947-951,956,共6页Recent Advances in Ophthalmology

基　　金：西安交通大学基金资助[编号:2020YJ(ZYTS)339]。

摘　　要：目的探究KIF11在高糖诱导的人视网膜微血管内皮细胞(HRMECs)损伤中的作用和机制。方法将正常培养的HRMECs随机分成四组:正常组(给予5 mmol·L^(-1)葡萄糖)、高糖组(给予30 mmol·L^(-1)葡萄糖)、高糖+si-NC组(转染100 nmol·L^(-1) si-NC后给予30 mmol·L^(-1)葡萄糖)、高糖+si-KIF11组(转染100 nmol·L^(-1) si-KIF11后给予30 mmol·L^(-1)葡萄糖)。Real-time PCR检测各组HRMECs的KIF11、VEGF和HIF-1αmRNA表达,Western blot检测KIF11、VEGF、HIF-1α和Wnt/β-catenin通路相关蛋白(β-catenin、cyclin D1和c-Myc)表达,CCK-8法检测细胞活力,Transwell检测迁移细胞数目。通过LiCl激活Wnt/β-catenin通路进一步确证KIF11是否通过该通路发挥作用。结果与正常组相比,高糖组中HRMECs细胞活力增加,迁移细胞数增多,KIF11、VEGF和HIF-1αmRNA和蛋白水平以及β-catenin、cyclin D1和c-Myc蛋白表达均上调(均为P<0.05);与高糖组相比,高糖+si-KIF11组中HRMECs细胞活力降低,迁移细胞数减少,KIF11、VEGF和HIF-1αmRNA和蛋白水平以及β-catenin、cyclin D1和c-Myc蛋白表达均下调(均为P<0.05);而高糖+si-NC组中上述指标与高糖组相比差异均无统计学意义(均为P>0.05)。与高糖+si-KIF11组相比,高糖+si-KIF11+LiCl组HRMECs细胞活力增加,迁移细胞数增多,VEGF和HIF-1α蛋白表达上调(均为P<0.05)。结论KIF11在高糖诱导的HRMECs中表达上调,下调其表达可通过抑制Wnt/β-catenin通路缓解高糖诱导的HRMECs损伤。Objective To investigate the role and mechanism of kinesin family member 11(KIF11)in the high glucose-induced injury of human retinal microvascular endothelial cells(HRMECs).Methods The normally cultured HRMECs were randomly divided into four groups:control group(5 mmol·L^(-1) glucose was administered),high glucose group(30 mmol·L^(-1) glucose was administered),high glucose+small interfering negative control(si-NC)group(transfected with 100 nmol·L^(-1) si-NC prior to 30 mmol·L^(-1) glucose addition),and high glucose+si-KIF11 group(transfected with 100 nmol·L^(-1) si-KIF11 prior to 30 mmol·L^(-1) glucose addition).Real-time PCR was used to detect the messenger ribonucleic acid(mRNA)expressions of KIF11,vascular endothelial growth factor(VEGF)and hypoxia-inducible factor-1α(HIF-1α)in HRMECs of each group.Western blot was used to determine the expression levels of KIF11,VEGF,HIF-1αand Wnt/β-catenin pathway-related proteins(β-catenin,cyclin D1,and c-Myc).Cell viability was analyzed by Cell Counting Kit-8.Migrated cell number was detected by Transwell.Additionally,the Wnt/β-catenin pathway was activated by LiCl to further confirm whether KIF11 functioned through this pathway.Results Compared with the control group,the cell viability and the number of migrated cells were increased,the mRNA and protein expression levels of KIF11,VEGF and HIF-1αand the expression levels ofβ-catenin,cyclin D1 and c-Myc were up-regulated in the high glucose group(all P<0.05).Compared with the high glucose group,the cell viability and the number of migrated cells were decreased,the mRNA and protein expression levels of KIF11,VEGF and HIF-1αand the expression levels ofβ-catenin,cyclin D1 and c-Myc were inhibited in the high glucose+si-KIF11 group(all P<0.05).However,there was no significant difference in the above indexes between the high glucose+si-NC and high glucose groups(all P>0.05).Compared with the high glucose+si-KIF11 group,the cell viability,the number of migrated cells and protein expression levels of VEGF and HIF-

关 键 词：KIF11 高糖 人视网膜微血管内皮细胞 WNT/Β-CATENIN通路 

分 类 号：R774[医药卫生—眼科]