高氧暴露对早产鼠AEC Ⅱ生长增殖的影响及CGRP的干预作用
Roles of calcitonin gene-related peptide on growth of premature rat type Ⅱ alveolar epithelial cells exposed to hyperoxia
付红敏 1许峰 1黄波 1方芳 1黄栋 1匡凤梧1
作者信息
- 1. 重庆医科大学儿童医院PICU室,重庆,400014
- 折叠
摘要
目的:探讨降钙素基因相关肽(CGRP)对600 mL/L氧暴露早产鼠肺泡Ⅱ型上皮细胞(AEC Ⅱ)生长增殖的影响.方法:原代分离培养孕19 d早产鼠AECⅡ,随机分为4组:空气组、高氧组、高氧CGRP组、高氧CGRP拮抗剂组.空气组,高氧组分别在氧体积分数为210 mL/L的空气和600 mL/L的氧气中暴露24 h;高氧CGRP组在暴露前加入CGRP;高氧CGRP拮抗剂组在暴露前同时加入CGRP和CGRP受体拮抗剂,再置于氧体积分数为600 mL/L的氧气中培养24 h.采用MTr比色法和流式细胞术测定细胞增殖能力和细胞周期;逆转录聚合酶链反应和免疫荧光技术测定表面活性蛋白C(SPC)的mRNA及蛋白表达.结果:与空气组比较,高氧组细胞存活率下降,G0/G1期细胞比例增高,G2/M及S期细胞比例降低,SPC mRNA及SPC蛋白表达低下(P均<0.01).而CGRP干预可促进高氧暴露AECⅡ的增殖能力,使S及G2/M期细胞增多,并提高AECⅡ的SPC mRNA及SPC蛋白表达水平.结论:600 mL/L氧暴露24 h可抑制早产鼠AEC Ⅱ增殖分化,而CGRP可促进AECH生长,部分解除高氧对AECⅡ的抑制作用.
Abstract
AIM: To explore the influence of 600 mL/L oxygen and the effects of calcitonin gene-related peptide ( CGRP) on the growth of type Ⅱ alveolar epithelial cells ( AEC Ⅱ ) isolated from premature rat lungs in vitro. METHODS: AEC Ⅱ were isolated from 19 d fetal rat lungs and cultured for 12 h to attach. AEC Ⅱ were then randomly divided into four groups: Air group, hyperoxia group, hyperoxia plus CGRP group, and hyperoxia plus CGRP and CGRP(8-37) group. AEC Ⅱ of air group and hyperoxia group were exposed to 210 mL/L or 600 mL/L oxygen respectively for 24 h while CGRP was added in hyperoxia plus CGRP group and CGRP and CGRP( 8-37 ) ( CGRP receptor antagonist) were added in hyperoxia plus CGRP and CGRP (8-37) group before they were placed into chamber of 600 mL/L oxygen. Cell proliferation ability was determined by MTT assay and cell cycles were examined by flow cytometry. The mRNA levels and protein contents of surfactant protein C ( SPC) in the four groups were measured by RT-PCR and immunofluorescence respectively. RESULTS: Exposure to 600 mL/L oxygen for 24 h inhibited AEC Ⅱ proliferation, increased cell proportion of G0/G1 phase with the decrease of cells in S and G2/M phases, and reduced the levels of mRNA and proteins of SPC. CGRP of 10 -8M promoted AEC Ⅱ proliferation, enhanced cell proportions of S and G2/M phases, and increased the expression of SPC mRNA and protein. CONCLUSION: Exposure to 600 mL/L oxygen for 24 h inhibits AEC Ⅱ growth, proliferation and differentiation whereas CGRP partially counteracts the inhibitory effects of hyperoxia on AEC Ⅱ and plays a protective role against hyperoxia-induced lung injury.
关键词
高氧/肺泡Ⅱ型上皮细胞/降钙素基因相关肽/婴儿,早产Key words
hyperoxia/Type Ⅱ alveolar epithelial cell/calcitonin gene-related peptide/infant/premature引用本文复制引用
基金项目
国家自然科学基金面上项目(30670931)
重庆市自然科学基金(2006BB5035)
出版年
2009
NETL
NSTL
万方数据