Wnt 5a在支气管肺发育不良模型中的作用机制探讨

Mechanism of Wnt 5a in bronchopulmonary dysplasia model

目的探究Wnt 5a在支气管肺发育不良(BPD)模型中的作用机制。方法(1)细胞水平:将A549细胞分为对照组(CO_(2)体积分数5%)和BPD组(O_(2)体积分数85%)。在高氧48 h后采用Western blot方法检测肺表面活性物质相关蛋白质C(SPC)、水通道蛋白5(AQP5)和Wnt 5a蛋白水平,细胞增殖检测试剂盒检测细胞增殖能力变化。(2)动物水平:将新生大鼠随机分为对照组(O_(2)体积分数21%)和BPD组(O_(2)体积分数85%)。高氧7 d后取肺组织,采用HE染色评估BPD动物模型病理变化;采用Western blot检测肺组织中Wnt 5a、SPC及AQP5变化。(3)细胞水平:进一步在BPD细胞模型中添加Wnt 5a抑制剂,验证Wnt 5a对SPC、AQP5蛋白及细胞增殖能力的影响。结果与对照组比较,BPD细胞模型中SPC和AQP5表达量降低,Wnt 5a表达量升高,细胞增殖能力降低。添加Wnt 5a抑制剂后SPC、AQP5蛋白表达升高,A549细胞增殖能力恢复。BPD动物模型中出现明显的肺组织肺泡数量减少、肺泡面积增大等病理表现;SPC、AQP5、Wnt 5a蛋白表达变化同BPD细胞模型趋势一致。结论Wnt 5a在BPD的发生发展中被异常激活,深入探索Wnt 5a在BPD中的作用机制可能会为BPD的防治提供新思路。

Objective To explore the mechanism of Wnt 5a in the rat model of broncho-pulmonary dysplasia(BPD).Methods(1)Cell level,A549 cells were divided into the two groups:the control group(carbon dioxide volume fraction 5%)and the BPD group(oxygen volume fraction 85%).After 48 hours of hyperoxia,the levels of pulmonary surfactant associated protein C(SPC),aquaporin 5(AQP5)and Wnt 5a proteins were detected by Western blot assay,and the change of cell proliferation ability was detected by cell proliferation detection kit.(2)Animal level,newborn rats were randomly divided into the two groups:the control group(oxygen volume fraction 21%)and the BPD group(oxygen volume fraction 85%).After7 days of hyperoxia,lung tissue was collected.HE staining was used to evaluate the pathological changes of BPD animal model,and Western blot assay was used to detect changes of Wnt 5a,SPC and AQP5 proteins in lung tissue.(3)Cell level,furthermore,Wnt 5a inhibitor was added to BPD cell model to verify the effect of Wnt 5a on SPC,AQP5 protein and cell proliferation.Results Compared with the control group,the expression levels of SPC and AQP5 decreased,the expression of Wnt 5a increased,and the ability of cell proliferation decreased in BPD cell model.The expression levels of SPC and AQP5 protein were recovered after cells were treatment with Wnt 5a inhibitor,and the proliferation ability of A549 cells was also recovered.The number of alveoli decreased and the area of alveoli increased in the BPD group.The expression levels of SPC,AQP5 and Wnt 5a protein were consistent with the teand of BPD cell model.Conclusion activated in the occurrence and development of BPD.In-depth exploration of the mechanism of Wnt 5a in BPD may provide new ideas for the prevention and treatment of BPD.