摘要
目的探讨Caveolae对降钙素基因相关肽(CGRP)抑制过氧化氢(H2O2)诱导的人脐静脉内皮细胞(HUVECs)损伤作用的影响及其机制。方法体外培养HUVECs,分别用CGRP和/或H2O2处理细胞,光学显微镜观察细胞形态学和密度的变化;流式细胞术观察细胞的增殖和周期分布;β-环糊精(β-CD)用于破坏caveolae结构;Western-blot检测caveolin-1表达。结果与正常组比较,CGRP组细胞密度增加,S期细胞数明显增多,H2O2组细胞密度降低,S期细胞数明显减少;CGRP预孵育细胞能抵抗H2O2引起的细胞数目减少;β-CD剥夺胆固醇,破坏caveolae结构,HUVECs形态学发生改变,但CGRP诱导的细胞密度和细胞增殖进一步提升,恢复胆固醇后,该作用被取消。与对照组比较,CGRP组细胞caveolin-1表达水平降低(P<0.05);H2O2组细胞caveolin-1水平上升(P<0.05),给予CGRP预孵育后,能显著逆转H2O2诱导的caveolin-1表达(P<0.05);β-CD破坏caveolae结构后,增强CGRP下调HUVECs caveolin-1表达的作用(P<0.05),增加胆固醇Caveolae结构有所恢复且该作用被削弱。结论 Caveolae结构完整性对CGRP保护H2O2损伤HUVECs有一定影响,破坏caveolae结构能增强CGRP对H2O2损伤的HUVECs的增殖作用,其机制可能与增强CGRP下调氧化应激导致的caveolin-1的表达有关。
Objective To study the role of caveolae on the protective effect of cacitonin gene-related peptide（ CGRP） on human umbilical vein endothelial cells（ HUVECs） injury induced by oxidative stress. Methods HUVECs were cultured in vitro,and treated with CGRP or / and hydrogen peroxide（ H2O2）,the distribution of cell cycles and proliferation of cells were observed by flow cytometry. The density and morphology of cells were observed by optical microscope,β-cyclodextrin（ β-CD） was used to injured the structure of caveolae,expression of caveolin-1 was measured by Western blot. Rethods Compared with control group,treatment of CGRP increased proliferation index（ PI） and the number of S phase cells which were inhibited by H2O2; Destruction of caveolae structure could increase the density of cells and change the cells shape; The expression of caveolin-1 was down-regulated by CGRP but up-regulated by H2O2,respectively. Pretreatment with CGRP,the up-regulation of caveolin-1 induced by H2O2 was significantly reverted（ P〈0. 05）. Pretreatment of β-cyclodextrin with cells,the expression of caveolin-1 induced by CGRP was furtherly decressed. Conclusions the destruction of caveolae could increased the proliferative effect of CGRP on HUVECs induced by H2O2,the mechanism involve in change of the expression of caveolin-1.
出处
《中南医学科学杂志》
CAS
2015年第2期132-136,共5页
Medical Science Journal of Central South China
基金
国家自然科学基金(30572192)
湖南省青年科学基金课题(13JJ4121)
