藤黄酸通过CYLD/NF-κB信号通路抑制高糖环境下RPE细胞上皮-间充质转化的作用  

Inhibition effect of gambogic acid on the epithelial-mesenchymal transition of retinal pigment epithelium cells via cylindromatosis/nuclear factor-kappaB signaling pathway under high glucose circumstance

作　　者：陈晶 庞东渤 CHEN Jing;PANG Dongbo(Department of Ophthalmology,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,Liaoning Province,China;Heilongjiang Sengong General Hospital,Harbin 150000,Heilongjiang Province,China)

机构地区：[1]锦州医科大学附属第一医院眼科,辽宁省锦州市121000 [2]黑龙江省森工总医院,黑龙江省哈尔宾市150000

出　　处：《眼科新进展》2022年第12期937-941,共5页Recent Advances in Ophthalmology

基　　金：辽宁省自然科学基金项目(编号:2019-ZD-0601)。

摘　　要：目的基于去泛素化酶圆柱瘤蛋白/核因子κB(CYLD/NF-κB)通路,研究藤黄酸(GA)对高糖环境下视网膜色素上皮(RPE)细胞上皮-间充质转化(EMT)的作用。方法体外培养ARPE-19细胞,高糖诱导,实验分为NC组(含5.5 mmol·L^(-1)葡萄糖),HG组(含30 mmol·L^(-1)葡萄糖),HG+不同剂量GA组(2μmol·L^(-1)、4μmol·L^(-1)、8μmol·L^(-1) GA分别预处理RPE细胞1 h,加30 mmol·L^(-1)葡萄糖)。CCK-8检测各组RPE细胞增殖活力,免疫荧光染色检测RPE细胞中α平滑肌肌动蛋白(α-SMA)、去泛素化酶圆柱瘤蛋白(CYLD)表达;Western blot检测RPE细胞中α-SMA、CYLD、磷酸化核转录因子κB(p-NF-κB)蛋白表达。结果与NC组相比,HG组RPE细胞出现明显增殖,RPE细胞中α-SMA、p-NF-κB蛋白表达均增加,CYLD蛋白表达减少(均为P<0.01);与HG组相比,HG+不同剂量GA组RPE细胞增殖均明显受到抑制,RPE细胞中α-SMA、p-NF-κB蛋白表达均减少,CYLD表达均增加(均为P<0.01),且均呈剂量依赖性。结论GA可抑制高糖环境下RPE细胞EMT,其抑制作用与上调CYLD,抑制NF-κB活化有关。Objective To study the effects of gambogic acid(GA)on the epithelial-mesenchymal transition(EMT)of retinal pigment epithelium(RPE)cells in high glucose environment via the deubiquitinating enzyme cylindromatosis/nuclear factor-kappaB(CYLD/NF-кB)signaling pathway.Methods The adult retinal pigment epithelium-19(ARPE-19)cells were cultured in vitro in a high-glucose medium.The normal control group(NC group,containing 5.5 mmol·L^(-1) glucose),high glucose group(HG group,containing 30 mmol·L^(-1) glucose),and HG+various doses of GA groups(the RPE cells were pre-treated with 2μmol·L^(-1) GA,4μmol·L^(-1) GA or 8μmol·L^(-1) GA for 1 h,and then added with 30 mmol·L^(-1) glucose)were set.The proliferation and viability of RPE cells in all groups were evaluated by Cell Counting Kit-8,the expression levels ofα-smooth-muscle-actin(α-SMA)and CYLD were detected by immunofluorescence staining,and the expression levels ofα-SMA,CYLD,and phosphorylation nuclear factor-kappaB(p-NF-κB)were measured by Western-blot.Results Compared with the NC group,in the HG group,the RPE cells proliferated significantly,the expressions ofα-SMA and p-NF-κB were up-regulated,and the CYLD protein expression was decreased(all P<0.01).Under the various doses of GA intervention,the proliferation of RPE cells was significantly inhibited,the expressions ofα-SMA and p-NF-κB decreased,and the CYLD expression increased(all P<0.01)in a dose-dependent manner as compared to the HG group.Conclusion Under high glucose circumstances,GA can inhibit the EMT of RPE cells by inducing CYLD expression and attenuating NF-кB activation.

关 键 词：藤黄酸 上皮-间充质转化 高糖 视网膜色素上皮细胞 去泛素化酶圆柱瘤蛋白 

分 类 号：R774[医药卫生—眼科]