miR-10调控Notch通路对缺氧环境诱导的视网膜神经节细胞损伤的影响  

Effect of miR-10 Regulation of Notch Pathway on Hypoxia-Induced Retinal Ganglion Cell Injury

作　　者：黄一凡 李珊珊 叶建荣[2] HUANG Yifan;LI Shanshan;YE Jianrong(Ophthalmology and Optometry,Binzhou Medical College,Binzhou,Shandong 256603,China;Department of Anesthesiology,The First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830054,China)

机构地区：[1]滨州医学院,山东滨州256603 [2]新疆医科大学第一附属医院麻醉科,新疆乌鲁木齐830054

出　　处：《转化医学杂志》2024年第10期1728-1732,共5页Translational Medicine Journal

基　　金：国家自然科学基金委员会资助项目(82060581)。

摘　　要：目的探索miR-10调控细胞跨膜受体蛋白1(Notch)通路对缺氧环境诱导的视网膜神经节细胞损伤的影响。方法将RGC-5细胞随机分为3组:对照组、miR NC组、miR-10组。miR NC组、miR-10组的RGC-5细胞经缺氧处理24 h后,分别转染miR NC、miR-10 mimic,持续处理24 h。通过MTT实验检测各组RGC-5细胞增殖活性,原位末端标记法分析各组RGC-5细胞凋亡情况,酶联免疫吸附测定法分析各组RGC-5细胞培养基中炎症因子[肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6]水平,双荧光素酶报告基因法分析miR-10与Notch1的靶向关系;蛋白免疫印迹法分析各组RGC-5细胞中Notch通路蛋白表达水平。结果与对照组比较,miR NC组RGC-5细胞增殖活性降低,凋亡率升高,TNF-α、IL-1β、IL-6水平升高,Notch1、发状分裂相关增强子1(Hes1)蛋白表达降低(均P<0.05)。与miR NC组比较,miR-10组RGC-5细胞增殖活性增加,凋亡率下降,TNF-α、IL-1β、IL-6水平降低,Notch1、Hes1蛋白表达增加(均P<0.05)。miR-10与Notch1具有靶向结合域。结论上调miR-10能够减轻缺氧环境诱导的视网膜神经节细胞损伤,该过程与miR-10靶向调控Notch通路相关。Objective To explore the effect of miR-10 on hypoxia-induced retinal ganglion cell(RGC)damage by regulating the Notch pathway.Methods RGC-5 cells were randomly divided into three groups:control group,miR NC group,and miR-10 group.RGC-5 cells in the miR NC group and miR-10 group were treated with hypoxia for 24hours.RGC-5 cells were transfected with miR NC and miR-10 mimic,respectively,for 24 hours.The viability of RGC-5 cells was assessed using the MTT assay.The levels of inflammatory factors in the culture medium of RGC-5 cells were analyzed by enzyme-linked immunosorbent assay.The apoptosis of RGC-5 cells was analyzed by in situ end labeling.The targeting relationship between miR-10 and Notch1 was analyzed by dual-luciferase reporter gene assay.The protein expression levels of Notch pathway proteins in RGC-5 cells were analyzed by Western blot.Results Compared with the control group,the proliferation activity of RGC-5 cells in the miR NC group was decreased,the apoptosis rate was increased,the levels of TNF-α,IL-1βand IL-6 were increased,and the expression of Notch1 and hairy and enhancer of split 1(Hes1)protein was decreased(all P<0.05).Compared with the miR NC group,the proliferation activity of RGC-5 cells in the miR-10 group was increased,the apoptosis rate was decreased,the levels of TNF-α,IL-1βand IL-6 were decreased,and the expression of Notch1 and Hes1 protein was increased(all P<0.05).miR-10 has a targeted binding domain with Notch1.Conclusion Up-regulation of miR-10 can reduce hypoxia-induced retinal ganglion cell damage,and this process is related to the regulation of the Notch pathway by miR-10.

关 键 词：miR-10 HES1 NOTCH1 视网膜神经节细胞 缺氧 增殖 炎症 凋亡 

分 类 号：R774.1[医药卫生—眼科]