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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 investigate whether calcitonin generelated peptide (CGRP) can promote the survival of type Ⅱ aveolar epithelial cells (AEC Ⅱ) under high oxygen concentration and its possible Wnt signaling mechanism. Methods Primary fetal AECⅡ fetal rat embryos were divided into three groups: air group, hyperoxia group, hyperoxia + CGRP group and hyperoxia + CGRP + hCGRP8-37 group. Cell proliferation was measured by MTT assay and cell death was detected by flow cytometry. The expression of Wnt7b, β-catenin and p-β-catenin were detected. Results Compared with the air group, the number of dead cells increased significantly (P <0.01), and the cell proliferation activity was significantly inhibited (P <0.01). CGRP significantly decreased the number of dead cells after hyperoxia exposure P <0.01), and attenuated the inhibitory effect of hyperoxia on cell proliferation (P <0.01). The expression of Wnt7b and β-catenin decreased significantly (P <0.01) and the expression of p-β-catenin increased significantly after hyperoxia exposure (P <0.01). Compared with hyperoxia exposure group, -catenin protein expression increased, while p-β-catenin expression decreased, with a statistically significant difference (P <0.01). Conclusion CGRP can reduce hyperoxia-induced cell death, attenuate the inhibitory effect of hyperoxia on cell proliferation and improve the survival of AECⅡ. Wnt7b / β-catenin pathway may be involved in the regulation of CGRP.