敷胸膏对流感病毒肺炎大鼠肺组织JNK,MKK4表达的影响

Effect of Fuxiong Cream on Expressions of JNK and MKK4 in Lung Tissue of Rats with Influenza A Virus Pneumonia

目的:探讨清肺通络敷胸膏对流感病毒肺炎大鼠肺组织中c-Jun氨基末端激酶(JNK),丝裂原活化蛋白激酶激酶4(MKK4)表达的影响,并探讨其作用机制。方法:幼龄Wistar大鼠随机分为正常组、模型组、敷胸膏高、中、低剂量组(21.0,10.5,5.25 g·kg-1),每组10只。除正常组外,其余各组采用甲型流感病毒鼠肺适应株A/FM/47(H1N1)0.1 m L滴鼻建立流感病毒肺炎模型,造模成功后各治疗组给予不同剂量敷胸膏治疗5 d。苏木素-伊红(HE)染色观察各组大鼠肺组织病理变化,免疫组化检测大鼠肺组织中JNK及MKK4的蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)进一步检测肺组织中JNK及MKK4 mRNA表达水平。结果:HE染色结果提示,与模型组比较,敷胸膏高、中、低剂量组能明显改善流感病毒肺炎大鼠肺组织损伤程度,随剂量增大而逐渐增强。免疫组化与Resl-time PCR结果显示,与正常组比较,模型组JNK,MKK4蛋白及mRNA表达显著增强(P<0.05);与模型组比较,敷胸膏高、中、低剂量组JNK,MKK4蛋白及mRNA表达均显著减弱(P<0.05)。结论:敷胸膏对流感病毒肺炎大鼠的保护作用机制可能与下调MAPKs信号通路中JNK,MKK4的表达有关。

Objective： To explore the effect of Fuxiong cream on expressions of c-Jun n-terminal kinase（ JNK） and mitogen-activated protein kinase kinase 4（ MKK4） in lung tissue of rats with influenza A virus（ IV）pneumonia. Method： Fifty young rats were randomly divided in the control group,the model group,the high,medium and low-dose Fuxiong cream groups（ 21. 0,10. 5,5. 25 g·kg-1）,with 10 rats in each group. Except for the control group,the IV pneumonia model was induced by nose dripping with 0. 1 m L rat lung adapted strain of influenza virus A/FM/47（ H1N1） in the remaining groups. After the successful modeling,the rats received Fuxiong cream for 5 days. HE staining was used to observe lung pathological histology. The expressions of JNK and MKK4 in rat lung tissue were detected by using immunohistochemical method. The mRNA expression levels of JNK and MKK4 in lung tissue were further detected by using Real-time PCR. Result： HE staining showed that the high,medium and low-dose Fuxiong cream groups could significantly improve the acute lung tissue injury degree in rats compared with the model group,and the curative effect gradually enhanced with the increase of the dose.Immunohistochemical and Real-time PCR results showed that compared with the control group,in the model group,expressions of JNK and MKK4 were obviously increased（ P〈 0. 05）. Compared with model group,in high,medium and low-dose Fuxiong cream groups,expressions of JNK and MKK4 were obviously decreased（ P〈 0. 05）.Conclusion： The protective effect of Fuxiong cream against IV pneumonia in rats may be associated with downregulation of JNK and MKK4.