中国实用眼科杂志
中國實用眼科雜誌
중국실용안과잡지
CHINESE JOURNAL OF PRACTICAL OPHTHALMOLOGY
2014年
6期
797-802
,共6页
冷炫%胡洁%宋新%田景毅%吕林
冷炫%鬍潔%宋新%田景毅%呂林
랭현%호길%송신%전경의%려림
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
乙酰肝素酶%血管內皮生長因子%組蛋白3乙酰化%糖尿病視網膜病變
을선간소매%혈관내피생장인자%조단백3을선화%당뇨병시망막병변
Heparanase%Vascular endothelial growth factor%Histone H3 acetylation%Diabetic retinopathy
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.
目的 研究高糖誘導人視網膜血管內皮細胞(HRECs)中組蛋白3乙酰化(H3ac)、乙酰肝素酶(HPA)和血管內皮生長因子(VEGF)錶達的相關性,明確一種或多種能夠參與HPA上調VEGF錶達的H3乙酰化賴氨痠殘基.方法 實驗研究.在中山大學中山眼科中心將該實驗分為兩組,正常組和高糖組(葡萄糖含量30mmol/L).免疫熒光染色法觀察兩組H3乙酰化賴氨痠殘基9(H3K9ac)、H3K14ac、H3K18ac、H3K27ac和H3K56ac在HRECs的錶達定位.蛋白質免疫印跡法(Western blot)檢測兩組間HPA、VEGF和各H3乙酰化賴氨痠殘基的蛋白錶達變化.結果 免疫熒光染色結果顯示,高糖組HRECs細胞覈中各H3乙酰化賴氨痠殘基的熒光較正常組增彊.Western blot檢測結果示,與正常組相比,高糖組中HPA、VEGF、H3K9ac和H3K14ac的蛋白錶達均有明顯升高,差異有統計學意義(PHPA=0.016,PVEGF=0.014,PH3K9ac=0.027,PH3K14ac=0.045).結論 高糖誘導的HRECs中,H3乙酰化在HPA參與VEGF基因轉錄調控的過程中髮揮作用,併且從蛋白水平證實乙酰化髮生在H3K9和H3K14.
목적 연구고당유도인시망막혈관내피세포(HRECs)중조단백3을선화(H3ac)、을선간소매(HPA)화혈관내피생장인자(VEGF)표체적상관성,명학일충혹다충능구삼여HPA상조VEGF표체적H3을선화뢰안산잔기.방법 실험연구.재중산대학중산안과중심장해실험분위량조,정상조화고당조(포도당함량30mmol/L).면역형광염색법관찰량조H3을선화뢰안산잔기9(H3K9ac)、H3K14ac、H3K18ac、H3K27ac화H3K56ac재HRECs적표체정위.단백질면역인적법(Western blot)검측량조간HPA、VEGF화각H3을선화뢰안산잔기적단백표체변화.결과 면역형광염색결과현시,고당조HRECs세포핵중각H3을선화뢰안산잔기적형광교정상조증강.Western blot검측결과시,여정상조상비,고당조중HPA、VEGF、H3K9ac화H3K14ac적단백표체균유명현승고,차이유통계학의의(PHPA=0.016,PVEGF=0.014,PH3K9ac=0.027,PH3K14ac=0.045).결론 고당유도적HRECs중,H3을선화재HPA삼여VEGF기인전록조공적과정중발휘작용,병차종단백수평증실을선화발생재H3K9화H3K14.
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.
