论文部分内容阅读
目的:探讨缺氧复氧对血管内皮细胞分泌一氧化氮(NO)及NO合酶(NOS)活力的影响及辛伐他汀的干预作用,并分析其产生作用的可能机制。方法:体外培养人脐静脉内皮细胞株ECV304,使用自制的缺氧小室对ECV304进行缺氧复氧处理。第1实验:仅进行缺氧复氧处理。第2实验:不同浓度的辛伐他汀(0.1、1.0、10.0μmol/L)以及10.0μmol/L辛伐他汀加0.2mmol/L甲羟戊酸预处理ECV30424h后,再行缺氧2h复氧2h处理。硝酸还原酶法及化学比色法分别检测培养液上清中NO的含量及NOS活力。结果:缺氧2h、复氧2、4hECV304分泌NO与对照组相比明显减少,复氧2、4hNOS活力与对照组相比明显下降;经1.0、10.0μmol/L浓度辛伐他汀预处理ECV304后再行缺氧复氧处理则NO含量及NOS活力较对照组明显增加;同时用辛伐他汀和甲羟戊酸预处理后NO含量和NOS较对照组差异无统计学意义。结论:缺氧复氧应激下血管内皮细胞分泌NO功能下降、NOS活力降低;辛伐他汀可以阻止因缺氧复氧刺激导致的NO减少及NOS活力下降,该作用可被甲羟戊酸逆转。
Objective: To investigate the effects of hypoxia and reoxygenation on the secretion of nitric oxide (NO) and nitric oxide synthase (NOS) in vascular endothelial cells and the intervention of simvastatin, and to analyze the possible mechanism. Methods: Human umbilical vein endothelial cell line ECV304 was cultured in vitro, and the ECV304 cells were subjected to hypoxia / reoxygenation treatment using home-made hypoxic chambers. The first experiment: only hypoxia and reoxygenation treatment. In the second experiment, different doses of simvastatin (0.1,1.0,10.0μmol / L) and 10.0μmol / L simvastatin plus 0.2mmol / L of mevalonic acid pretreated ECV30424h, followed by reoxygenation for 2h deal with. Nitric acid reductase method and chemical colorimetric method were used to detect the content of NO and the activity of NOS in the supernatant of culture medium. Results: After 2 hours of hypoxia, the NO secreted by ECV304 in reoxygenation 2,4 h significantly decreased compared with that in control group, and the activity of 2,4 hNOS in reoxygenation group decreased significantly compared with that in control group. After ECV304 was pretreated with 1.0 and 10.0 μmol / L simvastatin Then hypoxia and reoxygenation NO content and NOS activity increased significantly compared with the control group; at the same time with simvastatin and mevalonate pretreatment NO content and NOS compared with the control group no significant difference. CONCLUSIONS: The function of NO secreted by vascular endothelial cells is decreased and the activity of NOS is decreased under hypoxia-reoxygenation stress. Simvastatin can prevent the decrease of NO and the decrease of NOS activity induced by hypoxia-reoxygenation stimulation, which can be reversed by mevalonate .