清肺透邪方对肺炎支原体肺炎小鼠Notch信号通路的影响

Effect of Lung-Clearing and Pathogens-Expelling Recipe on Notch Signaling Pathway in Mycoplasma Pneumoniae Pneumonia Mice

【目的】观察清肺透邪方对肺炎支原体肺炎小鼠肺组织Notch信号通路的影响,探讨其抗肺炎支原体感染的分子机制。【方法】将144只SPF级BABL/c小鼠随机分成6组,即正常组,模型组,西药组,中药大、中、小剂量组,每组24只。除正常组外,采用鼻腔接种肺炎支原体菌株法将其余5组小鼠复制肺炎支原体肺炎模型。造模结束后,西药组小鼠给予阿奇霉素片(剂量为2.6 mg/mL)灌胃5 d,5 d后继续灌胃等体积生理盐水,中药大、中、小剂量组分别予清肺透邪方颗粒剂(剂量分别为1.9500、0.9750、0.4875 g/mL)灌胃,正常组与模型组小鼠给予等体积生理盐水灌胃。灌胃0.2 mL/d,共14 d。于给药后第3、7、10、14天进行取材。采用苏木素-伊红(HE)染色法观察小鼠肺组织病理表现,采用蛋白免疫印迹(Western Blot)法检测小鼠肺组织Notch1、Notch2、Notch3蛋白表达水平,采用定量逆转录聚合酶链反应(qRT-PCR)法测定小鼠肺组织Notch1、Notch2、Notch3 m RNA表达水平。【结果】小鼠肺炎支原体感染后镜下符合肺炎表现;西药组,中药大、中剂量组小鼠肺部炎症明显改善,且清肺透邪方作用呈时间、剂量依赖性。模型组小鼠肺组织Notch1、Notch2蛋白及mRNA表达水平较正常组显著升高(P<0.05),西药组,中药大、中剂量组Notch1、Notch2蛋白及mRNA表达水平均较模型组降低(P<0.05),且清肺透邪方作用呈时间、剂量依赖性。各组Notch3蛋白及m RNA表达比较,差异均无统计学意义(P>0.05)。【结论】清肺透邪方可以减轻肺炎支原体肺炎小鼠肺部炎症,其机制可能与抑制肺组织Notch通路中Notch受体的表达有关。

Objective To observe the effect of Lung-clearing and Pathogens-expelling Recipe on Notch signaling pathway in mice with mycoplasma pneumoniae pneumonia(MPP),so as to explore the molecular mechanism of resisting mycoplasmal pneumoniae infection.Methods One hundred and forty-four SPF BABL/c mice were randomly divided into 6 groups,namely normal group,model group,western medicine group,big-,middleand small-dose Chinese medicine groups,24 mice in each group.Apart from the normal group,the mice in the other 5 groups were induced into MPP model by intranasal inoculation of mycoplasma pneumoniae.After modeling,the western medicine group was given intragastric administration of azithromycin tablets(at the dosage of 2.6 mg/mL)for 5 days,followed by intragastric administration of the same volume of normal saline,the big-,middle-and small-dose Chinese medicine groups were given intragastric administration of Lung-clearing and Pathogens-expelling Recipe granules(at the dosage of 1.9500,0.9750,0.4875 g/mL,respectively),and the normal group and model group were given intragastric administration of the same volume of normal saline.Intragastric administration at 0.2 mL was performed once a day,lasting 14 days.Samples were taken on day 3,7,10,14 after medication.The pathological feature of lung tissues was observed by haematoxylin-eosin(HE)staining method.The protein expression levels of Notch1,Notch2 and Notch3 were detected by Western blotting method.The mRNA expression levels of Notch1,Notch2 and Notch3 were determined by quantitative reverse transcription polymerase chain reaction(qRT-PCR).Results After mycoplasma pneumoniae infection,the pathological changes of pulmonary inflammation were seen under microscope,while the pulmonary inflammation was reduced in the western medicine group and big-and middle-Chinese medicine groups,the effect of Lungclearing and Pathogens-expelling Recipe being in time-and dose-dependent manner.The protein and mRNA expression levels of Notch1 and Notch2 in the model group were significantly higher than those in the normal group(P<0.05),the protein and mRNA expression levels of Notch1 and Notch2 in the western medicine group and big-and middle-Chinese medicine groups were lower than those in the model group(P<0.05),the effect of Lung-clearing and Pathogens-expelling Recipe being in time-and dose-dependent manner.There was no significant difference in the protein and mRNA expression level of Notch3 among various groups(P>0.05).Conclusion Lung-clearing and Pathogens-expelling Recipe is effective for relieving pulmonary inflammation in MPPmice,and it’s mechanisms were related with inhibiting Notch receptor expression in lung Notch pathway.