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目的:研究枳实提取物及其药效组分橙皮苷和新橙皮苷对氧化低密度脂蛋白(oxidized low density lipoprotein,Ox-LDL)损伤的人脐静脉内皮细胞(human umbilical vein endothelial cells line,HUVEC)细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)表达和一氧化氮(nitric oxide,NO)释放的影响。方法:体外培养HUVEC,50μg/mL ox-LDL制造HUVEC损伤模型。以MTS染色法检测细胞毒性确定用药浓度。细胞ELISA法测定细胞表面ICAM-1的含量,试剂盒测定细胞培养上清液中NO含量。结果:①枳实提取物小于等于2 mg/mL时,橙皮苷浓度小于等于0.03125 mg/mL时,新橙皮苷浓度小于等于0.25 mg/mL时,HUVEC存活率分别大于80%。②2.0 mg/mL和1.0 mg/mL两个浓度的枳实提取物、15.625μg/mL的橙皮苷和0.2500 mg/mL新橙皮苷对ox-LDL诱导的HUVEC的ICAM-1表达有显著抑制作用。③2.0 mg/mL枳实提取物显著提高ox-LDL诱导的HUVEC和正常HUVEC培养液中的NO含量;7.813μg/mL、15.625μg/mL和31.250μg/mL 3个浓度的橙皮苷能显著提高ox-LDL诱导的HUVEC培养液中的NO含量,31.250μg/mL的橙皮苷能促进正常HUVEC的NO释放;0.2500 mg/mL和0.1250 mg/mL 2个浓度的新橙皮苷能显著提高ox-LDL诱导的HUVEC培养液中的NO含量。结论:枳实提取物及其药效组分橙皮苷、新橙皮苷能抑制ox-LDL诱导的HUVEC的ICAM-1表达,促进ox-LDL诱导的HUVEC的NO释放。
AIM: To investigate the effects of hesperidin and hesperidin on oxidative low density lipoprotein (Ox-LDL) injury of human umbilical vein endothelial cells line, HUVEC) on the expression of intercellular adhesion molecule-1 (ICAM-1) and the release of nitric oxide (NO). Methods: HUVECs were induced by HUVEC and 50μg / mL ox-LDL in vitro. MTS staining cytotoxicity determination of drug concentration. The cell surface ICAM-1 content was measured by ELISA and the content of NO in cell culture supernatant was determined by kit. Results: ① When the hesperidin concentration is less than or equal to 2 mg / mL, the survival rate of HUVEC is greater than 80% when the hesperidin concentration is less than or equal to 0.03125 mg / mL and the hesperidin concentration is less than or equal to 0.25 mg / mL. ② The ICAM-1 expression of ox-LDL-induced HUVEC in the two concentrations of 2.0 mg / mL and 1.0 mg / mL of Fructus Aurantii, 15.625 μg / mL hesperidin and 0.2500 mg / mL neohesperidin were Significant inhibitory effect. (3) 2.0 mg / mL citrus aurantium extract significantly increased NO content in ox-LDL-induced HUVECs and normal HUVEC culture medium; 7.813μg / mL, 15.625μg / mL and 31.250μg / mL 3 concentrations of hesperidin The content of NO in HUVECs induced by ox-LDL was significantly increased. Hesperidin 31.250μg / mL could promote the release of NO from normal HUVECs. The concentrations of 0.2500mg / mL and 0.1250mg / mL neohesperidin significantly increased Improve the ox-LDL-induced HUVEC culture medium NO content. CONCLUSION: Hesperidin and neohesperidin can inhibit the expression of ICAM-1 in ox-LDL-induced HUVEC and promote the release of NO in HUVEC induced by ox-LDL.