银杏叶提取物在高糖环境下对人视网膜微血管内皮细胞的作用及可能机制  

Effects of Ginkgo biloba extract on human retinal capillary endothelial cells under high glucose condition

作　　者：王俞方[1] 彭辉灿[1] 燕建军[1] 陈磊[1] 

机构地区：[1]南华大学第二附属医院眼科,湖南省衡阳市421001

出　　处：《眼科新进展》2013年第9期822-825,共4页Recent Advances in Ophthalmology

摘　　要：目的研究不同浓度的银杏叶提取物(Ginkgo biloba extract,GBE)在高糖环境下对人视网膜微血管内皮细胞(human retinal capillary endothelial cells,HRCEC)的作用及对基质金属蛋白酶2(matrix metalloproteinase-2,MMP2)和核转录因子κB(nuclear transcription factor kappaB,NF-κB)p65表达的影响,为糖尿病视网膜病变的药物防治提供新的思路和理论依据。方法体外原代培养HRCEC,并通过免疫细胞化学方法鉴定;用GBE作用于原代培养的HRCEC,实验分为空白对照组、低糖对照组、高糖对照组、高糖+不同浓度GBE(12.5 mg·L-1、25.0 mg·L-1、50.0 mg·L-1、100.0 mg·L-1)组,分别使用MTT比色法观察其对细胞增殖的影响,使用免疫细胞化学及Western blotting检测药物作用前后HRCEC中MMP2及NF-κB p65的表达。结果 TT比色法结果显示,用不同浓度的GBE处理高糖(25.0 mmol·L-1)环境下HRCEC,作用24 h、48 h和72 h,高糖+不同浓度GBE对HRCEC增殖有促进作用,并存在时间和浓度的依赖性。高糖+不同浓度(12.5 mg·L-1、25.0 mg·L-1、50.0 mg·L-1、100.0 mg·L-1)GBE组作用24 h细胞增殖率分别为(16.4±0.7)%、(30.6±0.8)%、(41.3±1.5)%、(52.6±1.8)%,作用48 h细胞增殖率分别为(21.8±1.2)%、(36.2±1.3)%、(48.0±1.4)%、(62.7±1.6)%,作用72 h细胞增殖率分别为(29.9±0.5)%、(46.2±0.8)%、(61.2±0.8)%、(73.4±1.6)%。随着药物浓度的逐渐提高和作用时间的增加,细胞存活率逐渐提高,而细胞抑制率逐渐下降。用不同浓度的GBE处理HRCEC,作用72 h,行免疫细胞化学检测MMP2的表达,运用Western blotting检测NF-κB p65的表达,发现随着GBE药物浓度的增高,高糖环境下HRCEC中的MMP2、NF-κB p65表达量逐渐减少(P<0.05)。结论 GBE可以保护高糖环境下HRCEC并下调MMP2、NF-κB p65的表达,可用于糖尿病视网膜病变的防治。Objective To investigate the effects of Ginkgo biloba extract （GBE） of different concentrations on human retinal capillary endothelial ceils （HRCEC） and expressions of matrix metalloproteinase-2 （MMP2） and nuclear transcription factor kap- paB κBp65 （NF-κB 1）65 ） under high glucose condition, and to provide new ideas and theoretical basis for medication prevention and treatment for diabetic retinopathy. Methods Primary culture of HRCEC was carried out in v/tro, and the cells were i- dentified using immunocytochemistry. Apart from blank control group, HRCECs were randomly divided into six experimental groups：low glucose control group, high glucose control group,high glucose ＋ 12.5 mg·L^-1 GBE group,high glucose ＋ 25.0 mg·L^-1 GBE group,high glucose ＋50.0 mg·L^-1GBE group and high glucose ＋ 100.0 mg·L^-1 GBE group. Cell proliferation of each group was measured using MTr assay. The ex- pressions of MMP2 and NF-κB p55 in HRCEC before and after GBE administration were detected using immunocytochemistry and Western blotting. Results According to MTT assay, the cell proliferation in all high glucose （25.0 mmol·L^-1） ＋ GBE groups were oromoted after an action time of 24 hours,48 hours and 72 hours, and there was atime-and With an action time of 24 hours,cell proliferation rates of all high glucose ＋GBE（ 12.5 nag·L^-1,25.0 mg ·L^-1,50.0 mg ·L^-1,100.0 mg ·L^-1） groups were（16.4±0.7）% ,（30.6 ±0.8）% ,（41.3 ± 1.5）% , （52. 6 ± 1.8）% ,respectively. That were（21.8 ± 1.2）% , （36.2 ± 1.3）% , （48.0 ~ 1.4）% and（62.7 ± 1.6）% with an action time of 48 hours;and were（29.9 ±0.5）% , （46.2 ±0.8）% , （61.2 ± 0. 8）% and（73.4 ± 1.6）% with an action time of 72 hours. With the rise of GBE concentration and action time, ceil survival ratio gradually increased, while ceil inhibition ratio gradually decreased. After an action time of 72 hours,the expressions of MMP2 and NF-κB p65 in all high glucose ＋ GBE groups were detected by immu

关 键 词：银杏叶提取物 人视网膜血管内皮细胞 基质金属蛋白酶 核转录因子ΚB 

分 类 号：R774.1[医药卫生—眼科]