糖尿病视网膜病变中组蛋白3乙酰化参与乙酰肝素酶调控VEGF基因转录的初步研究  

Preliminary study of histone H3 acetylation involved in heparanase regulation of VEGF gene in diabetic retinopathy

作　　者：冷炫[1] 胡洁[1] 宋新[1] 田景毅[1] 吕林[1] 

机构地区：[1]中山大学中山眼科中心眼科学国家重点实验室,广州510060

出　　处：《中国实用眼科杂志》2014年第6期797-802,共6页Chinese Journal of Practical Ophthalmology

基　　金：国家自然科学基金（81271009）

摘　　要：目的 研究高糖诱导人视网膜血管内皮细胞（HRECs）中组蛋白3乙酰化（H3ac）、乙酰肝素酶（HPA）和血管内皮生长因子（VEGF）表达的相关性,明确一种或多种能够参与HPA上调VEGF表达的H3乙酰化赖氨酸残基.方法 实验研究.在中山大学中山眼科中心将该实验分为两组,正常组和高糖组（葡萄糖含量30mmol/L）.免疫荧光染色法观察两组H3乙酰化赖氨酸残基9（H3K9ac）、H3K14ac、H3K18ac、H3K27ac和H3K56ac在HRECs的表达定位.蛋白质免疫印迹法（Western blot）检测两组间HPA、VEGF和各H3乙酰化赖氨酸残基的蛋白表达变化.结果 免疫荧光染色结果显示,高糖组HRECs细胞核中各H3乙酰化赖氨酸残基的荧光较正常组增强.Western blot检测结果示,与正常组相比,高糖组中HPA、VEGF、H3K9ac和H3K14ac的蛋白表达均有明显升高,差异有统计学意义（PHPA=0.016,PVEGF=0.014,PH3K9ac=0.027,PH3K14ac=0.045）.结论 高糖诱导的HRECs中,H3乙酰化在HPA参与VEGF基因转录调控的过程中发挥作用,并且从蛋白水平证实乙酰化发生在H3K9和H3K14.Objective To investigate the relationships among expressions of Histone 3 acetylation,heparanase （HPA） and vascular endothelial growth factor （VEGF） in high glucose human retinal endothelial cells （HRECs） and the modulating styles of acetylated H3 lysine （H3ac） involved in HPA-mediated increasing expression of VEGE Methods Normal group and high glucose group （30mmol/L） were studied.The locations and expressions of H3 acetylated lysine 9 （H3K9ac）,H3K14ac,H3K18ac,H3K27ac and H3K56ac in HRECs were respectively measured by immunofluorescence and western blot.Results Immunofluoresence results showed that distributions of 5 kinds of H3ac were more intense in nucleus of high glucose HRECs compared with normal group.The expressions of HPA,VEGF,H3K9ac and H3K14ac in high glucose HRECs were statistically higher than that in normal group in western blot （P 〈0.05）.Conclusions H3 acetylation （H3K9ac and H3K14ac） might connect with HPA-mediated transcriptional regulation of VEGF gene.

关 键 词：乙酰肝素酶 血管内皮生长因子 组蛋白3乙酰化 糖尿病视网膜病变 

分 类 号：R734.2[医药卫生—肿瘤]