钩藤总生物碱和芥子碱硫氰酸盐组分配伍对血管内皮细胞的变化作用和机制

The Effect and Mechanism of Component Compatibility of Rhynchophylla Total Alkaloids and Sinapine Cyanide Sulfonate on Change of Vascular Endothelial Cells

目的研究中药组分钩藤总生物碱、芥子碱硫氰酸盐及其配伍对肿瘤坏死因子α(TNF-α)导致的血管内皮细胞的变化作用和机制。方法体外培养大鼠血管内皮细胞,以TNF-α建立血管内皮细胞炎症模型。采用扫描电镜技术观察细胞形态的变化,采用ELISA测定细胞分泌内皮素1(ET-1)、血管细胞黏附分子1(VCAM-1)和细胞间黏附分子1(ICAM-1),RT-PCR测定ICAM-1、VCAM-1和ET-1的mRNA表达,硝酸还原酶法测定细胞分泌血管内皮舒张因子一氧化氮(NO)水平。结果 TNF-α(5μg/L)可在体外诱导血管内皮炎症模型。钩藤总生物碱与芥子碱硫氰酸盐配伍可改善细胞形态结构,降低ET-1、ICAM-1和VCAM-1分泌,与钩藤总生物碱、芥子碱硫氰酸盐差异无显著性(P>0.05);提高细胞NO分泌,优于钩藤总生物碱、芥子碱硫氰酸盐(P<0.05);下调细胞ICAM-1 mRNA表达,优于钩藤总生物碱、芥子碱硫氰酸盐(P<0.05);下调细胞VCAM-1和ET-1 mRNA表达,优于芥子碱硫氰酸盐(P<0.05)。结论钩藤总生物碱、芥子碱硫氰酸盐及其配伍具有保护TNF-α致血管内皮细胞炎症的作用。

Aim To study the effect and mechanism of rhynchophylla total alkaloids, sinapine cyanide sulfonate and its component compatibility on change of vascular endothelial cells induced by the tumor necrosis factor-α（TNF-α）. Methods Rat vascular endothelial cells were cultured in vitro, and vascular endothelial cell injury model was established by TNF-α. Then changes of cell morphology were observed before and after treatment; intercellular adhesion molecule-1 （ICAM-1）, vascular cell adhesion molecule-1 （ VCAM-1 ） and rat endothelin-1 （ ET-1 ） were determined by Elisa method, mRNA of ICAM-1, VCAM-1 and ET-1 by RT-PCR method and the level change of endothelium-derived relaxing factor （NO） by nitrate reductase method. Results The vascular endothelial cell injury model could be established in vitro successfully by TNF-ct （5 μg/L）. Compared with TNF-α group, treated groups could improve vascular endothelial cell morphology, lower the secretion of ICAM-1 ,VCAM-1 and ET-1. Compared with rhynchophylla total alkaloids group and sinapine cyanide sulfonate group, the component compatibility group was better on heightening the secretion of NO, down-regulating the expression of ICAM-1 mRNA （P〈0.05）. On down-regulating the expression of mRNA of VCAM-1 and ET-1, the rhynchophylla total alkaloids group and the component compatibility group were better than the sinapine cyanide sulfonate group（P 〈0.05）. Conclusion The component compatibility of rhynchophylla total alkaloids and sinap- ine cyanide sulfonate has the function of protecting rat vascular endothelial cell injury induced by TNF-α.