芹菜素在风轮菜改善高糖所致血管内皮细胞损伤中的作用  

Effect of apigenin on amelioration of Clinopodium chinense for vascular endothelial cell injury induced by high glucose

作　　者：王立英[1] 吴斐华[1] 马龙[1] 李伟生[1] 梁敬钰[2] 马世平[1] 

机构地区：[1]中国药科大学中药药理教研室,江苏南京211198 [2]中国药科大学天然药物化学教研室,江苏南京211198

出　　处：《中草药》2014年第20期2946-2952,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金青年基金资助项目(81102862);国家科技重大专项重大新药创制(2011ZX09307-002-02);江苏省高校青蓝工程资助项目;中国药科大学中央高校基本科研业务费专项基金培育项目(02410018)

摘　　要：目的通过特异剔除芹菜素(AP),研究AP在风轮菜活性部位(CCE)改善高糖诱导的人脐静脉内皮细胞(EA.hy926)损伤中的作用地位。方法应用Sephadex LH-20凝胶柱色谱法从CCE中特异剔除AP,制备剔除AP的CCE(CCEAP-)。将培养的内皮细胞随机分为5组:对照组、高糖模型组、CCE组、CCEAP-组和AP组。以MTT法检测细胞存活率,流式细胞仪检测胞内活性氧(ROS)水平,通过Hoechst染色,荧光显微镜观察细胞凋亡形态,以Annexin V-FITC染色,流式细胞仪检测细胞凋亡率,以分光光度法检测细胞内caspase-3的活性,并用Western blotting检测Bax的表达情况。结果 CCE组和AP组均能显著提高高糖损伤的内皮细胞存活率,降低高糖诱导的内皮细胞胞内ROS水平、内皮细胞凋亡率、caspase-3的活性及Bax的表达。而CCEAP-组对高糖所致内皮细胞损伤的改善作用不明显。结论 AP是风轮菜改善高糖所致内皮细胞损伤的重要成分之一,其改善高糖诱导内皮细胞凋亡作用与抗氧化应激有关。Objective In this study, we specifically deleted apigenin（AP） from the active fraction of Clinopodium chinense（CCE）, and aimed at identifying the effect of AP on how CCE exerted its amelioration on high glucose-induced injury in human umbilical vein endothelial cells（EA.hy926）. Methods By using Sephadex LH-20 gel column chromatography, AP was specifically deleted from CCE, and the apigenin-depleted sample of CCE（CCEAP-）was obtained. The cultured endothelial cells were divided into five groups： normal control group, high glucose model group, CCE group, CCEAP- group, and AP group. The cell viability was assayed by MTT assay. Flow cytometry was used to measure the intracellular reactive oxygen species（ROS）. Morphology of cell apoptosis was determined by fluorescence microscopy with Hoechst staining. The rate of apoptosis was measured by flow cytometry staining with AnnexinV-FITC. Caspase-3 activity was measured using caspase-3 colorimetric assay kit. The expression of Bax was detected by Western blotting. Results CCE and AP could significantly improve the cell viability, reduce the generation of intracellular ROS in EA.hy926 induced by high glucose. Meanwhile, CCE and AP can reduce the ratio of apoptosis, activity of caspase-3, and expression of Bax, while CCE knocked out apigenin（CCEAP-） had a slight improvement on the high glucose-induced endothelial cell injury. Conclusion AP is one of the principal components improving the high glucose-induced endothelial cell injury, and its anti-apoptosis effect may be related to anti-oxidative stress.

关 键 词：风轮菜 芹菜素 高糖 内皮细胞 凋亡 特异性剔除 

分 类 号：R285.5[医药卫生—中药学]