Qingzao Jiufei Decoction And Its Separate Components Regulate NLRP3 Inflammasome-related Factors In Mice Infected With Mycoplasma Pneumoniae

Purpose:Establishment of Mycoplasma pneumoniae pneumonia(MPP)model in BALB/c mice infected with Mycoplasma pneumoniae.To investigate the effect of Qingzao Jiufei Decoction and its decomposition on the expression of inflammatory corpuscle-related factors of NLRP3 in MPP mice induced by MP,and to further clarify its targets,Qingzao Jiufei Decoction and its decomposition were used to intervene mycoplasma-infected mice.Material and method:120 SPF Balb/c mice were selected and randomly divided into normal group,model group,Qingzao Jiufei Decoction group,Decomposing prescription I group,Decomposing prescription II group and Azithromycin group with 20 mice in each group.Except for the normal group,MPP mice were established by using Mycoplasma droplets from the nasal cavity of mice to lower respiratory tract infection mechanism.After the establishment of the model,each group was given 15 g/kg of corresponding prescriptions by gavage once a day for14 days.Azithromycin group was given azithromycin 0.09g/kg,once a day,intragastric administration for 3 consecutive days,withdrawal for 4 days,two cycles,and distilled water intragastric administration during withdrawal.Normal group and model group were given 0.3m L distilled water per stomach.The samples were taken after 4 hours of fasting and water deprivation.Samples were taken on the 3rd,7th and 14 th day after infection.Six mice were randomly executed in each group to observe the pathological changes of lung tissue in mice.Lung index was measured by weighing method.The expression of NLRP3 protein in lung tissue was detected by immunohistochemistry SP.The expression of NLRP3 and Caspase-1 protein in lung tissue was detected by immunohistochemistry SP and Western blot,and the level of interleukin-1 beta in serum was detected by ELISA.Results:1.The pathological sections of lung tissue in normal mice were basically normal.The structure of alveoli,alveolar septum and alveolar sac was intact.No inflammatory cell infiltration was found around them.After MP infection,the structure of alveoli,alveolar septum and alveolar sac disappeared,a large number of inflammatory cells infiltrated,and the wall of bronchioles became thicker in the model group,especially on the 7th day,and the inflammation gradually decreased on the 14 th day.After treatment,a small amount of inflammatory cells were observed in alveoli and the wall of bronchioles was thickened and alleviated in Qingzao Jiufei Decoction group.On the 3rd and 7th day after modeling,there was a significant improvement compared with model group(P < 0.05).The infiltration of inflammatory cells in alveolar tissue of mice in Decomposing prescription I group was reduced,and it was significantly improved on the 7th day after model establishment compared with that in Decomposing prescription I group(P < 0.05).Inflammatory infiltration in lung tissue of mice in Decomposing prescription II group was not significantly improved,and there was no significant difference between Decomposing prescription II group and model group(P >0.05).A small amount of inflammatory cell infiltration was observed in mice of azithromycin group.Compared with model group,the number of inflammatory cells decreased significantly(P < 0.05).2.After MP infection,the lung index of model mice was significantly higher than that of normal mice,reaching its peak on the 7th day(P < 0.05).After drug intervention,the lung index of mice in Qingzao Jiufei Decoction group was lower than that in model group on the3 rd,7th and 14 th day after treatment(P < 0.05).The mice in Decomposing prescription I group were significantly different from those in model group on the 3rd,7th and 14 th day after treatment(P < 0.05).Decomposing prescription II group and model group,on the 3rd day after treatment,there was no significant difference between the two groups(0.01 < P < 0.05).and significant differences between the two groups on the 7th and 14 th day after treatment(P< 0.05).The mice in azithromycin group were significantly different from those in model group on the 3rd,7th and 14 th day after treatment(P < 0.05).There was no significant difference in pulmonary index among Qingzao Jiufei Decoction group,Decomposing prescription I group and azithromycin group(P > 0.05).3.The expression of NLRP3 protein in lung tissue of mice was detected by SP immunohistochemical method.The results showed that the expression of NLRP3 in lung tissue of mice increased significantly 3,7 and 14 days after MP infection(P < 0.01).After drug intervention,the expression of NLRP3 was down-regulated in Qingzao Jiufei Decoction group,Decomposing prescription I group,Decomposing prescription II group and Azithromycin group(P < 0.05).Among them,Qingzao Jiufei Decoction group,Decomposing prescription I group and Decomposing prescription II group began to take effect on the 7th day(P < 0.05).4.The expression of NLRP3 protein in lung tissues of mice was detected by Western blot.The results showed that the expression of NLRP3 in lung tissues of mice increased significantly on the 3rd,7th and 14 th day after MP infection(P < 0.01).After drug intervention,the expression of NLRP3 was down-regulated in Qingzao Jiufei Decoction group,Decomposing prescription I group,Decomposing prescription II group and Azithromycin group(P <0.05).Among them,Qingzao Jiufei Decoction group,Decomposing prescription I group and Decomposing prescription II group began to take effect on the 7th day(P < 0.05).5.The expression of Caspase-1 protein in lung tissues of mice was detected by Western blot.The results showed that the expression of Caspase-1 protein in lung tissues of mice increased significantly 3,7 and 14 days after MP infection(P < 0.01).After drug intervention,the expression of Caspase-1 could be down-regulated in Qingzao Jiufei Decoction group,Decomposing prescription I group,Decomposing prescription II group and Azithromycin group(P < 0.05).4.Among them,Qingzao Jiufei Decoction group,Decomposing prescription I group and Decomposing prescription II group began to take effect on the 7th day(P < 0.05).6.On the 3rd,7th and 14 th day after MP infection,the expression of IL-1beta in serum of mice increased significantly(P < 0.01),reaching its peak on the 3rd day.After drug intervention,the expression of IL-1beta could be down-regulated in Qingzao Jiufei Decoction group,Decomposing prescription I group,Decomposing prescription II group and Azithromycin group(P<0.05).4.Among them,Qingzao Jiufei Decoction group,Decomposing prescription I group and Decomposing prescription II group began to take effect on the 3rd day(P < 0.05).Conclusion:1.Qingzao Jiufei Decoction can effectively alleviate pulmonary inflammation in MPP mice,and its effect is more significant in the late stage of MP infection.2.Qingzao Jiufei Decoction can effectively down-regulate the expression of Caspase-1,NLRP3 in lung tissue and IL-beta in serum of MP-infected mice,which may be the target of its therapeutic effect on MPP.3.Qingzao Jiufei Decoction for Decomposing prescription I is more effective than Decomposing prescription II.Its constituent drugs may play a major role in the prescription.