降钙素基因相关肽对高氧暴露胎鼠原代肺泡Ⅱ型细胞的保护作用及其Wnt信号机制  

Calcitonin gene-related peptide protects premature rat primary type Ⅱ alveolar epithelial cells against damage induced by hyperoxia

作　　者：王少华[1] 许峰[1] 党红星[1] 

机构地区：[1]重庆医科大学附属儿童医院PICU,重庆400014

出　　处：《第三军医大学学报》2012年第9期836-838,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金(30973218)~~

摘　　要：目的探讨降钙素基因相关肽(calcitonin generelated peptide,CGRP)是否可促进高氧损伤肺泡Ⅱ型上皮细胞(typeⅡalveolar epithelial cells,AECⅡ)的存活及其可能涉及的Wnt信号机制。方法 SD胎鼠原代AECⅡ分为空气组、高氧组、高氧+CGRP组、高氧+CGRP+hCGRP8-37组,采用MTT测定各组细胞增殖,流式细胞仪检测细胞死亡,Western blot检测Wnt7b、β-catenin及p-β-catenin表达。结果与空气组对照比较,高氧暴露24 h后死亡细胞数显著升高(P<0.01),细胞增殖活性明显受到抑制(P<0.01),CGRP干预可明显下调高氧暴露后死亡细胞数(P<0.01),减弱高氧对细胞增殖的抑制作用(P<0.01)。高氧暴露后Wnt7b及β-catenin蛋白表达水平明显降低(P<0.01),p-β-catenin表达水平显著增加(P<0.01),CGRP干预后,与单纯高氧暴露组相比Wnt7b、β-catenin蛋白表达水平增加,而p-β-catenin表达水平降低,具统计学差异(P<0.01)。结论 CGRP可减少高氧诱导的细胞死亡、减弱高氧对细胞增殖的抑制效应,改善AECⅡ存活,Wnt7b/β-catenin通路可能参与了CGRP这一调控过程。Objective To determine the effects of calcitonin gene related peptide（CGRP） and Wnt on type Ⅱ alveolar epithelial cells（AECⅡ） upon the exposure of hyperoxia.Methods AECⅡ were isolated and purified from premature rats and exposed to air,hyperoxia,hyperoxia＋CGRP（10-7 mol/L） and hyperoxia＋CGRP＋hCGRP8-37（10-6 mol/L） respectively.The proliferation of AECⅡ was measured by MTT cell proliferation assay.Cell apoptosis was measured by flow cytometry.The activation of Wnt7b,β-catenin and p-β-catenin was assessed by Western blot analysis.Results Cell proliferation was significantly decreased and apoptosis was increased upon the treatment of hyperoxia（P0.01）.Compared with hyperoxia exposure alone,additional CGRP treatment promoted cell proliferation and reduced the apoptosis of AECⅡ（P0.01）.Hyperoxia exposure resulted in significant decrease of Wnt7b and β-catenin（P0.01） and increased expression of p-β-catenin（P0.01）,while CGRP treatment reversed above results significantly（P0.01）.Conclusion CGRP decreases cell death and attenuates the inhibited proliferation of hyperoxia exposure,and improves the survival of AECⅡ exposed to hyperoxia,which might be associated with the activation of Wnt7b/β-catenin.

关 键 词：降钙素基因相关肽 高氧 肺泡Ⅱ型细胞 肺损伤 WNT信号 

分 类 号：R322.35[医药卫生—人体解剖和组织胚胎学] R329.26[医药卫生—基础医学]