CGRP抑制AngⅡ诱导HUVECs的损伤及其ERK1/2信号通路

CGRP Protect the HUVECs Injuried by AngII and the ERK1/2 Signal Pathway

目的:探索降钙素基因相关肽(CGRP)对经血管紧张素II(AngII)损伤的人脐静脉内皮细胞(HUVECs)的保护作用且CGRP与细胞外信号调节激酶(ERK1/2)的关系。方法:不同浓度的CGRP、AngII处理体外培养的HUVECs,噻唑蓝比色法检测HUVECs活力;流式细胞仪分析HUVECs凋亡率及其增殖指数;显微镜观察HUVECs的形态学变化;Western blot检测p-ERK1/2的表达。结果:AngII(0.1-100 nmol/L)浓度依赖性降低HUVECs的活力,而CGRP(0.1-1000 nmol/L)浓度依赖性增加HUVECs的活力;HUVECs增殖指数PI值受AngII、CGRP及PD98059(ERK1/2抑制剂)影响;AngII孵育HUVECs在第10min时ERK1/2磷酸化水平可达到最大;CGRP能抑制AngII诱导的HUVECs内ERK1/2磷酸化水平;CGRP8-37(CGRP受体拮抗剂)可部分减弱CGRP抑制ERK1/2磷酸化水平作用;PD98059(ERK1/2抑制剂)作用下,ERK1/2磷酸化水平显著降低,但是对细胞内总ERK1/2水平表达无明显影响。结论:CGRP可抑制AngII对HUVECs的损伤作用,可能与CGRP抑制信号通路ERK1/2有关。

Objective： To investigate the effect of CGRP on protecting the human umbilicalvein endothelial cells （HUVECs） from injuries by angiotensin Ⅱ （AngⅡ） in vitro, and to investigate its relationship with the activity of p-ERK1/2. Methods： HUVECs were cultured in vitro. The viability and cell cycle of cultured HUVECs were respectively estimated by MTT assay and Flow Cytometry （FCM）. The morphology of HUVECs was detected by microscope. Western blot was used to detect the effect ofpretreatment with CGRP on the expression of p-ERK1/2 induced by AnglI. Results： CGRP （0.1-1000 nmol/L） dose-dependently increased the viability of HUVECs. AngⅡ（0.1-100 nmol/L） dose-dependently decreased the viability of HUVECs. The value index （PI） decreased in HUVECs induced by AnglI, then increased by CGRP and PD98059. WB showed treat the HUVECs with AnglI regulated the expression of p-ERK1/2 and the effect was distinct at 10 minitus. CGRP down-regulated the expression of p-ERK1/2 which was partly attenuated by CGRP8-37. PD98059 also down-regulated the level ofp-ERK1/2. Conclusion： CGRP may protect HUVECs from AnglI induced injuries, the mechanism may be related with the ERK1/2 signal pathway.