化痰祛瘀疏肝法预防性干预对哮喘小鼠NGF-TrkA信号通路的影响

Effect of Preictal Phase Intervention of Phlegm-Stasis Resolving and Liver Soothing Method on NGF-TrkA Signaling Pathway of Murine Model of Asthma

目的:探讨化痰祛瘀疏肝法预防性干预对哮喘小鼠NGF-TrkA信号通路的影响。方法:以10%OVA/AL(OH)3致敏、5%OVA激发复制支气管哮喘小鼠模型,于激发前开始给药,至激发结束取材。用ELISA法检测BALF中NGF、TrkA浓度,Wstern blot法检测肺组织匀浆中NGF、TrkA蛋白表达。结果:模型组NGF、TrkA浓度明显升高(P均<0.01),NGF、TrkA蛋白表达明显上调(P<0.05,P<0.01);各处理组NGF、TrkA浓度均较模型明显降低(P均<0.01),两中药组NGF蛋白表达均明显低于模型组(P均<0.01),地米组与模型组无统计学意义(P>0.05),两中药组TrkA蛋白表达均明显低于模型组(P<0.05,P<0.01);两中药组NGF、TrkA浓度下调不及地米组(P均<0.01);PCF组NGF、TrkA蛋白表达与地米组相似(P均>0.05),SG-PCF组显著优于地米组(P均<0.05);两中药组NGF、TrkA浓度及蛋白表达均无统计学意义(P均>0.05)。结论:在发作期干预方式下,化痰祛瘀疏肝法对哮喘小鼠升高的NGF、TrkA浓度及上调的NGF、TrkA蛋白表达均有降低作用,上述作用总体与化痰祛瘀平喘法相似,优于地塞米松。提示化痰祛瘀疏肝法对哮喘小鼠神经源性炎症NGF-TrkA信号通路有较为确切的干预作用。

Objective：To observe the effects of preictal intervention of phlegm and stasis resolving and liver soothing method on NGFTrkA signaling pathway of neurogenic inflammation of murine model of asthma.Methods：The asthmatic BALB /c mice model were established through being sensitized by intra-peritoneal injection of 10% OVA /AL（OH） 3 solution,and activated by 5% ovalbumin solution.Intervention started before activation,and lung homogenate was obtained after activation.NGF and TrkA concentration in BALF were examined by ELISA,and the protein expression of NGF and TrkA concentration in lung tissue homogenate were detected by western blot.Results：In asthmatic model BALB /c mice,NGF and TrkA concentration significantly increased（P ＆lt; 0.01）,the protein expression significantly increased（P ＆lt; 0.05,P ＆lt; 0.01）.The NGF、TrkA concentration of all intervention groups decreased（P ＆lt; 0.01）,while DXM showed a better effect than the other two intervention groups（P ＆lt; 0.01）.The NGF protein expression in PCF and SGPCF group decreased significantly compared with model group（P ＆lt; 0.01）,while that of DXM group was similar to model group（P ＆gt; 0.05）.TrkA protein expression of PCF and SGPCF group were lowered（P ＆lt; 0.05,P ＆lt; 0.01）,PCF group had a similar effect on NGF and TrkA with DXM,while SGPCF showed a significantly better effect than DXM（P ＆lt; 0.05）,but the differences between PCF and SGPCF group had no statistical significance（P ＆gt; 0.05）.Conclusion：Preictal intervention of phlegm and stasis resolving and liver soothing method can decrease the increased concentration and protein expression of NGF and TrkA.The effect is generally similar to phlegm and stasis resolving and qi regulating method,and better than dexamethasone,which indicates that preictal intervention of phlegm and stasis resolving and liver soothing method has a definite effect on the NGF-TrkA pathway of neurogenic inflammation of murine model of asthma.