基于“肺通调水道”理论探究CFTR介导下鞘脂代谢失衡对慢性阻塞性肺疾病的调控机制

Based on the Theory of“ Lung Channel Regulation”, the Regulatory Mechanism of CFTR-MediatedImbalance of Infingolipid Metabolism on Chronic Obstructive Pulmonary Disease was Investigated

目的探讨CFTR调控下鞘脂代谢在COPD中的发病过程及作用途径,进一步阐释肺通调水道理论。方法烟熏法建立COPD小鼠模型,同时烟熏加CFTR激动剂建立CFTR模型,观察肺组织病理变化评估小鼠模型,采用LC-MS质谱检测模型血浆中鞘脂代谢产物Ceramide及sphingosine-1-phosphate表达,采用Western blot方法检测小鼠模型肺组织中Sphks、ASM、CFTR蛋白磷酸化水平,采用荧光定量PCR小鼠模型肺组织中Sphks、Smpd基因(ASM)、CFTRmRNA转录水平。结果COPD组、CFTR干预组中S1p表达均较对照组降低(P<0.05),CFTR干预组较COPD组表达高(P<0.05);CFTR、Sphk1蛋白磷酸化水平在COPD组及CFTR干预组均呈低表达,COPD组表达最低与对照组、CFTR干预组有差异(P<0.05),Sphk2在COPD组与对照组存在差异(P<0.05)。ASM在COPD组、CFTR干预组高于对照组(P<0.05)。CFTR mRNA在COPD组及CFTR干预组均较对照组低,COPD组与对照组存在差异(P<0.05),Sphk1 mRNA在对照组表达最高,且与COPD组、CFTR干预组均存在差异(P<0.05),SMPD1 mRNA在COPD组及CFTR干预组呈高表达,且与对照组有差异(P<0.05)。结论探讨肺通调水道功能失司在COPD疾病中的物质变化基础,揭示CFTR通过参与调控鞘脂代谢,从而影响COPD水液代谢的途径。

Objective The material basis and pathway of CFTR regulation of sphingolipid metabolism in COPD were discussed,and the theory of lung channel regulation was further elucidated.Methods The mouse model of COPD was established by smoking method,and the CFTR model was established by smoking plus CFTR agonist.The pathological changes of lung tissues were observed and the mouse model was evaluated.Ceramide and sphingosine-1-phosphate expression of sphingolipid metabolites in plasma of the model were detected by LC-MS mass spectrometry.Western blot was used to detect the phosphorylation levels of Sphks,ASM and CFTR proteins in the lung tissue of the mouse model.Quantitative fluorescence PCR was used to detect the mrna transcription levels of Sphks,Smpd and CFTR mRNA in the lung tissue of the mouse model.Results The expression of S1p in COPD group and CFTR intervention group was lower than that in control group(P<0.05),and the expression of S1P in CFTR intervention group was higher than that in COPD group(P<0.05).The protein phosphorylation levels of CFTR and Sphk1 were low in COPD group and CFTR intervention group,the lowest expression in COPD group was different from that in control group and CFTR intervention group(P<0.05),and Sphk2 was different in COPD group and control group(P<0.05).ASM in COPD group and CFTR intervention group was higher than that in control group(P<0.05).CFTR mRNA in COPD group and CFTR intervention group was lower than that in control group,and there were differences between COPD group and control group(P<0.05).Sphk1 mRNA expression was the highest in control group,and there were differences between it and COPD group and CFTR intervention group(P<0.05).SMPD1 mRNA was highly expressed in COPD group and CFTR intervention group,and was different from control group(P<0.05).Conclusion To explore the material changes of pulmonary aqueduct dysfunction in COPD diseases,and to reveal the pathway of CFTR affecting water and fluid metabolism in COPD by participating in the regulation of sphingolipid metabolism.