银杏双黄酮对高糖致微血管内皮细胞炎症反应的保护作用及机制研究  

Protective effect of ginketin on inflammatory response of microvascular endothelial cells induced by high glucose and itsmechanism

作　　者：郭丽 路青瑜[1,2] 张启云 杨付梅 孙黔云 GUO Li;LU Qin-yu;ZHANG Qi-yun;YANG Fu-mei;SUN Qian-yun(State Key Labof Functions and Applications of Medicinal Plants, Guizhou Medical University, Guiyang 550014,China;Key Lab of Chemistry for Natural Products, Guizhou Province and Chinese Acadamy of Science, Guiyang 550014, China)

机构地区：[1]贵州医科大学省部共建药用植物与功效与利用国家重点实验室 [2]贵州省中国科学院天然产物化学重点实验室,贵州贵阳550014

出　　处：《中国药理学通报》2020年第10期1463-1469,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(NoU1812403);贵州省科技计划及平台人才项目(No黔科合人才[2016]4018号,黔科合平台人才[2016]5625号,黔科合平台人才[2019]5702号)。

摘　　要：目的研究银杏双黄酮对高糖致人微血管内皮细胞(HMEC)炎症反应的干预作用及其可能的分子机制。方法MTT比色法检测细胞活力,采用ELISA检测细胞培养上清中可溶性黏附分子ICAM-1和炎症介质(IL-6、TNF-α)的含量变化;荧光免疫法和流式细胞仪测定HMEC细胞表面ICAM-1的表达;双荧光素酶报告基因检测NF-κB p65核内转录活性;Western blot检测NF-κB p65及p-NF-κB p65、Bax、Bcl-2的表达水平。结果HMEC暴露于高糖后,NF-κB信号通路活化,NF-κB p65磷酸化及核内转录活性明显上调,HMEC表达ICAM-1和TNF-α、IL-6明显升高,凋亡相关蛋白Bax/Bcl-2的比值出现上调。1×10^-5、1×10^-6、1×10^-7 mol·L^-1的银杏双黄酮对上述炎症反应相关指标的上调均有明显的干预作用,且呈现一定的量效关系。结论银杏双黄酮对高糖刺激HMEC炎症反应有明显的干预作用,其机制可能与抑制NF-κB信号通路活化,从而降低炎症反应水平有关。Aim To investigate the protective effect of ginketin on inflammatory response of microvascular endothelial cells induced by high glucoseand the underlying mechanism.Methods The cell viability of HMEC was detected by MTT assay.The content of soluble adhesion molecule ICAM-1 and inflammatory mediators(IL-6,TNF-α)in cell culture supernatant was measured by ELISA.The expression of ICAM-1 on the surface of cells was determined by immunofluorescence and flow cytometry,and the nuclear transcription activity of NF-κB p65was assessed by dual luciferase reporter assay.The protein expression levels of NF-κB p65,p-NF-κB p65,Bax and Bcl-2 were detected by western blot.Results After exposure HMEC to high glucose,the NF-κB signaling pathway was activated,the protein levels of NF-κB p65,p-NF-κB p65 and the activity of nuclear transcription were significantly up-regulated.The ICAM-1,TNF-αand IL-6 significantly increased,and the ratio of Bax/Bcl-2 was up-regulated.Ginketin at concentrations of 1×10^-5,1×10^-6 and 1×10^-7 mol·L^-1 showed significant intervention effects on the inflammatory response of HMEC mentioned above in a dose-dependent manner.Conclusions Ginketin possesses a significant intervention effect on inflammatory response of HMEC induced by high glucose,and its mechanism may be related to inhibiting the activation of the NF-κB inflammatory signaling pathway andreducing the level of inflammatory response.

关 键 词：银杏双黄酮 高糖 微血管内皮细胞 炎症反应 黏附分子 炎症介质 NF-κB信号通路 

分 类 号：R284.1[医药卫生—中药学] R322.12[医药卫生—中医学]