吴茱萸次碱对内皮细胞损伤的保护效应及机制

Protective effect of Rutaecarpine(RUT) on Human Umbilical Vein Endothelial Cell(HUVECs) Injury

目的研究吴茱萸次碱(rutaecarpine,Rut)的抗动脉粥样硬化(atherosclerosis,AS)内皮损伤作用和机制。方法在溶血性磷脂酰胆碱(lysoph-osphatidylcholine,LPC)诱导的内皮细胞损伤模型,研究Rut对内皮细胞的保护效应:用流式细胞术测定法检测细胞凋亡率、用MTT比色试验检测细胞活力、用全自动生化仪检测细胞培养基LDH水平、ELISA法检测TNF-α含量,荧光定量PCR检测CGRP mRNA水平,并用特异性阻断剂CAPZ(阻断VR1)或CGRP8-37(阻断CGRP受体),确证Rut的内皮保护效应与VR1/CGRP途径的关系。结果 Rut能有效地减弱LPC诱导的细胞活力下降、培养基LDH水平升高、TNF-α含量升高等内皮细胞损伤作用;Rut还呈现促进CGRP mRNA的表达等作用;并且以上作用会因为同时加入特异性阻断剂CAPZ(阻断VR1)或CGRP8-37(阻断CGRP受体)而被拮抗。结论 Rut具有抗AS内皮损伤作用,其机制与通过激活VR1促进CGRP的合适和释放有关。

Objective This study was designed to demonstrate the mechanism of antagonism effect of Rut against AS and endothelial injury（activation of VR1 to promote the release of CGRP）.Methods By using the model of LPC-induced endothelial cell injury,we investigate into the protective effect on endothelial cells of Rut,that include：using flow cytometer to detect cell apoptosis;using MTT colorimetric test for detection of cell viability;using a detection kit to measure the LDH level of cell culture medium;ELISA to detect TNF-α levels;quantitative PCR to detect the mRNA levels of CGRP;and using specific inhibitor CAPZ（block VR1） or CGRP8-37（blocking CGRP receptors）,to confirm the endothelial protective effect of Rut and the its role in VR1/CGRP pathway.Results Rut can effectively down regulate the factors of endothelial cell injury induced by LPC,which include the cell disability,high LDH level in cell medium and high TNF-α level;Rut show effects on mRNA expression of CGRP.These effects can be antagonized by adding specific inhibitor CAPZ（blockVR1） or CGRP8-37（blocking CGRP receptors）.Conclusion Rut have protective effect on AS and endothelial injury,which induced through the release of CGRP by activating VR1.