摘要
目的通过观察丹皮酚对同型半胱氨酸损伤的人脐静脉内皮细胞(HUVECs)一氧化氮合酶(eNOS)表达及一氧化氮(NO)水平影响,探讨丹皮酚对内皮细胞的保护作用及其抗动脉粥样硬化的分子机制。方法实验分为5组,分别为正常对照组、同型半胱氨酸损伤模型组(10 mmol/L同型半胱氨酸)、丹皮酚低剂量组(10 mmol/L同型半胱氨酸+0.15 mmol/L丹皮酚)、丹皮酚中剂量组(10 mmol/L同型半胱氨酸+0.3 mmol/L丹皮酚)、丹皮酚高剂量组(10 mmol/L同型半胱氨酸+0.6 mmol/L丹皮酚)。通过倒置显微镜观察各组HUVECs细胞形态学变化,通过MTT比色法测定细胞活力,采用RT-PCR法检测eNOS mRNA的表达,通过硝酸还原酶法测定NO水平。结果同型半胱氨酸损伤模型组的细胞呈团簇状,轮廓不清,出现较多细胞脱落的现象。丹皮酚各剂量组的细胞数量较多,细胞间隙逐渐变窄,形态趋于正常。与正常对照组比较,同型半胱氨酸损伤模型组细胞活力明显降低(P<0.01),eNOS mRNA的表达明显降低(P<0.01),NO水平明显降低(P<0.01)。与同型半胱氨酸损伤模型组比较,丹皮酚各剂量组的细胞活力明显升高(P<0.05),eNOS mRNA的表达明显增强(P<0.01),并呈现一定的剂量依赖性(P<0.05),NO水平明显升高(P<0.05)。结论丹皮酚对同型半胱氨酸损伤的内皮细胞具有保护作用,其机制可能为丹皮酚通过促进eNOS mRNA的表达,进而提高同型半胱氨酸损伤的内皮细胞内NO水平。
AIM To investigate the effects of paeonol on the expression of endothelial nitric oxide synthase（eNOS） and the level of nitric oxide（NO） in the human umbilical veins endothelial cells（HUVECs） injured by homocysteine,and their anti-atherosclerotic molecular mechanisms.METHODS HUVECs were randomly divided into five groups： normal control group,homocysteine injured model group（10 mmol/L homocysteine）,low-dose group of paeonol（10 mmol/L homouysteine＋0.15 mmol/L paeonol）,middle-dose group of paeonol（10 mmol/L homocysteine＋0.3 mmol/L paeonol）,and high-dose group of paeonol（10 mmol/L homocysteine ＋0.6 mmol/L paeonol）.The morphological changes of HUVECs were observed in each group respectively with inverted microscope.MTT assay was used to detect the cell viability.The expression of eNOS mRNA was tested by RT-PCR.The method of nitric acid reductase was used to measure the level of NO.RESULTS In homocysteine injured model group,the cells presented cluster shape,the cell contour became blurring,and some cells detached from the walls.In different doses of paeonol groups,the numbers of cells became more,the intercellular space became narrow,and the cell shape tended to be normal.Compared with the normal control group,the cell viability in homocysteine injured model group decreased obviously（P0.05）,eNOS mRNA expression decreased significantly（P0.01）,and the level of NO in the cultured supernate decreased significantly（P0.05）.Compared with homocysteine injured model group,the intercellular space became narrow,the cell shape tended to be normal,the cell viability increased significantly（P0.05）,the eNOS mRNA expression increased remarkably in a concentration-dependent manner（P0.05）,and the level of NO was remarkably higher in different doses of paeonol groups（P0.05）.CONCLUSION Paeonol plays a protective role in endothelial cells injured by homocysteine through the improvement of the gene expression of eNOS and increases the level of NO.
出处
《中成药》
CAS
CSCD
北大核心
2012年第12期2286-2289,2301,共4页
Chinese Traditional Patent Medicine
关键词
丹皮酚
同型半胱氨酸
人脐静脉内皮细胞
MTT
一氧化氮合酶
一氧化氮
paeonol
homocysteine
human umbilical veins endothelial cells
3-（4
5-dimethylthiazol-2-yl）-2
5-diphenyl tetrazolium bromide
endothelial nitric oxide synthase
nitric oxide
