摘要
目的探讨康柏西普(conbercept)对氯化钴(CoCl2)作用下体外培养SD大鼠视网膜Müller细胞系(rMC-1)ZnT8表达的影响。方法 rMC-1随机分为对照组(DMEM培养基培养)、低氧组(DMEM培养基培养+200μM氯化钴)及实验组(DMEM培养基培养+200μM氯化钴+10μg/mL的康柏西普处理)。结果 Müller细胞培养24 h后,谷氨酰胺合成酶(GS)强表达。低氧组ZnT8 mRNA、蛋白水平表达显著性降低(P<0.05),经康柏西普治疗后其表达显著增加。锌离子探针显示低氧组细胞内锌离子浓度增加、细胞凋亡显著增加,实验组都显著降低;同时外源性锌处理也会显著增加细胞凋亡,经康柏西普治疗后凋亡也显著降低。caspase-3蛋白水平呈现与细胞凋亡相同的结果,即低氧组显著增加,实验组显著降低。结论康柏西普可增加体外培养低氧环境下视网膜Müller细胞ZnT8的表达,抑制低氧诱导的细胞凋亡,降低caspase-3蛋白表达水平,促进细胞增殖。
Objective To investigate the effect of Conbercept on the expression of ZnT8 in retinal Müller cell line(rMC-1) of SD rats cultured in vitro with cobalt chloride. Methods rMC-1 was randomly divided into the control group(cultured with DMEM medium), the hypoxic group(cultured with DMEM medium + 200 μM cobalt chloride) and the experimental group(cultured with DMEM medium + 200 μM cobalt chloride + 10μg/mL Conbercept). Results Glutamine synthase(GS) was strongly expressed in Müller cells after culture for 24 hours. The expression of ZnT8 mRNA and protein in the hypoxic group decreased significantly(P<0.05), and increased significantly after treatment with Conbercept. Zinquin showed an increase in intracellular zinc ion concentration and cell apoptosis in the hypoxia group, but a decline in the experimental group. Meanwhile, exogenous zinc treatment also significantly increased cell apoptosis, which was significantly reduced after treatment with Conbercept. Level of Caspase-3 protein also increased significantly in the hypoxia group and decreased significantly in the experimental group, which was similar to apoptosis.Conclusions Conbercept can increase the expression of ZnT8 in retinal Müller cells cultured in hypoxic environment, inhibit apoptosis induced by hypoxia, reduce the expression of caspase-3 protein and promote cell proliferation.
作者
康道欢
史彩平
Kang Daohuan;Shi Caiping(Department of Ophthalmology,The Children's Hospital,Zhejiang University School of Medicine,national Clinical Research Center for Child Health,Hangzhou 310003,China)
出处
《临床眼科杂志》
2019年第6期552-556,共5页
Journal of Clinical Ophthalmology
