清燥救肺汤及其分解剂对肺炎支原体感染小鼠肺部炎症相关因子的影响

Effect of Qingzao Jiufei decoction and its decomposing agent on lung inflammation-related factors in mice infected with Mycoplasma pneumoniae

目的探讨清燥救肺汤(QJD)及其分解剂对肺炎支原体(MP)感染小鼠血清中TNF-α、INF-γ含量及肺组织中MPN372、P1、AQP5表达的影响,明确其抗MP感染的分子机制。方法将144只SPF级BABL/c小鼠随机分成正常组(A组)、模型组(B组)、QJD组(C组)、QJD分解剂I组(D组)、QJD分解剂II组(E组)及阿奇霉素组(F组),每组24只。除A组外,对其余5组小鼠进行MP感染模型处理,造模后分别予对应分组药物灌胃治疗,于造模后第3、7、10、14天进行取材。镜下观察小鼠肺组织病理切片,对小鼠肺组织病理炎症程度进行评分;并计算小鼠肺指数及干湿比;采用q PCR法对MPN372、P1基因含量进行测定;酶联免疫吸附试验(ELISA)测定各组小鼠血清TNF-α、INF-γ水平变化;Western blot法检测AQP5蛋白的表达。结果 MP感染后,镜下可见肺泡间隔增厚,细支气管壁增厚,大量炎症细胞浸润;其肺指数升高,干湿比降低;TNF-α、INF-γ的含量呈升高趋势,并于第7天达到峰值;AQP5的蛋白表达呈回落趋势,于第14天开始逐渐升高;与B组比较,从第7天开始,C组能够下调MPN372、P1及TNF-α的表达水平,上调INF-γ、AQP5的表达水平;其中D组的作用主要表现在下调MPN372、P1、TNF-α的表达,E组的作用主要表现在上调INF-γ、AQP5的表达。结论 QJD可以控制MP感染后肺部炎症,减少MP毒素MPN372的产生和P1黏附蛋白的表达;上调INF-γ、AQP5蛋白表达,下调TNF-α的表达,是其作用机制之一。

Objective To investigate the effects of Qingzao Jiufei Tang（ QJD） and its decomposing agent on the levels of TNF-α,INF-γ levels of TNF-α of Qingo,P1 and AQP5 in lung tissue of mice infected with Mycoplasma pneumoniae（ MP）,and to clarify its molecular anti-infective mechanism. Methods One hundred-forty-four SPF grade BABL/c mice were randomly divided into the normal group（ group A）,model group（ group B）,QJD group（ group C）,QJD decomposition agent I group（ group D） 、QJD decomposition group II Group（ group E） and azithromycin group（ group F）,with 24 rats in each group. Besides the group A,the other 5 groups of mice were treated with MP infection. After the mod-eling,the mice were given corresponding drugs by gastric gavage,and samples were obtained on the days 3,7,10,14 after the model was established. The lung tissue sections were examined by histopathology,and the degrees of inflammation in lung tissues in the mice were evaluated,and the lung index and the ratio of dry and wet lung weight ratio in the mice were calculated. The levels of MPN372 and P1 genes were determined by q PCR assay. The serum TNF-α and INF-γ levels in the mice were assessed by enzyme linked immunosorbent assay（ ELISA）. The expression of AQP5 protein was detected by Westernblot. Results After MP infection,the pathological examination revealed thickening of alveolar septum and bronchioles,and extensive inflammatory cell infiltration in the lung tissues. The lung index was increased and the ratio of dry and wet lung weight（ P〈0. 05）. TNF-α and INF-γ cedd and was increased,and reached the peak on the seventh day. The expression of AQP5 protein showed a downward tendency,and began to gradually increase on the 14 th day. Compared with the group B,the expression levels of MPN372,P1 and TNF-α in the group D were down-regulated,and the expression levels of INF-γ levein the group E were up-regulated from the 7 th day. Conclusions QJD can control pulmonary inflammation in mice after MP infection. The decrease of production of MP toxin MPN372 and the expression of adhesion protein P1,the up-regulation of expression of INF-γ and AQP5 proteins,and down-regulation of TNF-α expression are one of the mechanisms of its action.