摘要
目的:探讨circFAM73A靶向调控miR-140-3p对高糖诱导的人视网膜血管内皮细胞凋亡、迁移和炎症因子释放的影响及作用机制。方法:将人视网膜血管内皮细胞分别采用5.5 mmol/L葡萄糖(NG组)或25 mmol/L葡萄糖(HG组)处理,在HG组分别转染si-circRNA FAM73A和miR-140-3p inhibitors后,RT-qPCR检测circRNA FAM73A、miR-140-3p、TNF-α和IL-6表达水平;流式细胞术和Western blot分别检测细胞凋亡情况和凋亡相关分子Cleaved caspase 3的表达水平;Transwell检测细胞迁移情况;双荧光素酶报告实验与RIP检测circRNA FAM73A和miR-140-3p的靶向结合情况。结果:circFAM73A的表达水平在高糖处理人视网膜血管内皮细胞后显著升高,miR-140-3p的表达水平显著下降(均P<0.05);沉默circFAM73A能够显著抑制高糖诱导的人视网膜血管内皮细胞凋亡、迁移、炎症因子TNF-α和IL-6的释放(均P<0.05);低表达miR-140-3p能够显著促进高糖诱导的人视网膜血管内皮细胞凋亡、迁移、炎症因子TNF-α和IL-6的释放(均P<0.05);双荧光素酶报告和RIP实验证实circRNA FAM73A能够靶向结合miR-140-3p;共转染si-circRNA FAM73A和miR-140-3p inhibitors能够显著减弱低表达miR-140-3p对人视网膜血管内皮细胞凋亡、迁移和炎症因子释放的作用(均P<0.05)。结论:沉默circFAM73A能够通过靶向结合miR-140-3p抑制高糖诱导的人视网膜血管内皮细胞凋亡、迁移和炎症因子释放。
Objective:To investigate the effects and specific mechanisms of circFAM73A targeting regulation of miR-140-3p on high glucose-induced human retinal vascular endothelial cell apoptosis,migration and inflammatory cytokine release.Methods:Human retinal vascular endothelial cells were treated with 5.5 mmol/L glucose(control group,NG group)or 25 mmol/L glucose(high glucose group,HG group),and then transfected with si-circRNA FAM73A and miR-140-3p inhibitors in the HG group.The expression levels of circRNA FAM73A,miR-140-3p,TNF-α,and IL-6 were detected by RT-qPCR.Flow cytometry and Western blot were used to detect cell apoptosis and the expression levels of apoptosis-related molecule Cleaved caspase 3.Transwell assay was performed to detect cell migration.Dual-luciferase reporter assay and RNA immunoprecipitation(RIP)were used to detect the targeting binding of circRNA FAM73A and miR-140-3p.Results:The expression level of circFAM73A was significantly increased in human retinal vascular endothelial cells treated with high glucose(P<0.05),while the expression level of miR-140-3p was significantly decreased(P<0.05).Silencing circFAM73A could significantly inhibit high glucose-induced cell apoptosis,migration,and release of inflammatory cytokines TNF-αand IL-6 in human retinal vascular endothelial cells(all P<0.05).Low expression of miR-140-3p could significantly promote high glucose-induced cell apoptosis,migration,and release of inflammatory cytokines TNF-αand IL-6 in human retinal vascular endothelial cells(all P<0.05).Dual-luciferase reporter assay and RIP confirmed that circRNA FAM73A could target bind to miR-140-3p.Co-transfection of si-circRNA FAM73A and miR-140-3p inhibitors could significantly attenuate the effects of low expression of miR-140-3p on cell apoptosis,migration,and inflammatory cytokine release in human retinal vascular endothelial cells(all P<0.05).Conclusion:Silencing circFAM73A can inhibit high glucose-induced apoptosis,migration,and release of inflammatory cytokines in human retinal vascular endothelial cells by targeting binding to miR-140-3p.
作者
李伟华
安书杰
孟华
沙南希
王双勇
LI Weihua;AN Shujie;MENG Hua;SHA Nanxi;WANG Shuangyong(Department of Comprehensive Diagnosis and Treatment,Xijing Hospital of Air Force Medical University,Xi’an 710032,China)
出处
《陕西医学杂志》
CAS
2023年第11期1451-1457,共7页
Shaanxi Medical Journal
基金
国家自然科学基金资助面上项目(81200659)。
