山茱萸最佳配伍组分对高糖致ECV304细胞氧化损伤的保护作用  

The effect of the best compatibility of components in Fruetus Corni of protecting the oxidative damage of vascular endothelial cells cultured in high glucose medium

作　　者：许惠琴[1,2] 农伟虎[1,2] 刘成鼎[1] 李莉[1] 刘洪[1] 刘斌[1] 吴佳蕾[1] 

机构地区：[1]南京中医药大学药学院 [2]江苏省中药药效与安全性评价重点实验室,江苏南京210046

出　　处：《中国药理学通报》2011年第8期1077-1080,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81073111);江苏省自然科学基金资助项目(NoBK2001127);江苏省中医药局资助项目(山茱萸组分及其配伍防治早期糖尿病肾病的作用机理)

摘　　要：目的研究山茱萸最佳配伍组分(环烯醚萜苷、三萜酸和多糖以2∶1∶2配伍)对高糖培养的内皮细胞(human um-bilical vein endothelium cells,ECV304)氧化应激的影响。方法体外培养细胞,在细胞板内加入含30 mmol.L-1高糖的10%胎牛血清(fetal bovine serum,FBS)RPMI 1640培养液,加入空白、山茱萸最佳配伍组分大剂量、小剂量含药血清。同时设10%FBS RPMI 1640培养液作正常对照,川芎嗪作阳性对照。在37℃、5%CO2饱和湿度培养48 h后,取细胞上清液测定超氧化物歧化酶(SOD)、丙二醛(MDA),谷胱甘肽过氧化物酶(GSH-Px)。应用激光共聚焦显微镜检测进入细胞内2',7'-二氯荧光黄双乙酸酯(DCFH-DA)荧光强度来反映细胞内ROS含量。结果高糖组细胞上清液中SOD、GSH-Px活性降低,MDA、ROS含量增加(P<0.01),提示血管内皮细胞受损,抗氧化能力降低。而山茱萸最佳配伍组分能提高SOD、GSH-Px活力,降低ROS、MDA含量,与高糖组比较差异有显著性(P<0.05,0.01)。结论山茱萸最佳配伍组分能够保护血管内皮细胞,具有防治糖尿病血管病变的作用,其作用机制与抑制氧化应激有关。Aim To study the best compatibility components of Fructus Corni in culturing human umbilical vein endothelium cells（ECV304） from oxidant stress in high glucose.Methods Cells were cultured in vitro,the 10% fetal bovine serum（FBS） RPMI 1640 culture medium added to cell plates containing 30 mmol·L-1 high glucose,then the blank serum,the high dose drug-containing serum and the low dose drug-containing serum of the best compatibility of the Fructus Corni were added into them.Also 10% FBS RPMI 1640 medium was set as common control,and tetramethylpyrazine（TMP） as positive control.The cells were cultured at 37 ℃,5% CO2 saturated humidity,after 48 h,the supernatant fluid was taken to measure super oxide dismutase（SOD）,malondialdehydel（MDA） and glutathione peroxidase（GSH-Px）.The laser scanning confocal microscope was used to measure the fluorescence intensity of 2′,7′-dichlorofluorescin-diacetate（DCFH-DA） to reflect the content of reactive oxygen species（ROS） in cells.Results The cells cultured in high glucose group SOD,GSH-Px activity decreased,MDA,ROS content increased（P0.01）,showing endothelial cell damage and reduction of antioxidant capacity.The best compatibility of components of Fructus Corni could increase the activity of SOD and GSH-Px,and reduce the content of ROS and MDA,and it was significantly different compared to the high glucose groups（P0.05,0.01）.Conclusion The best compatibility components of Fructus Corni can protect vascular endothelial cells and have the function of controlling diabetes vascular lesions which is related with inhibitting oxidant stress.

关 键 词：山茱萸最佳配伍组分 ECV304 SOD MDA GSH-PX 氧化应激 

分 类 号：R282.71[医药卫生—中药学] R284.1[医药卫生—中医学]