Effects Of Zhiyan Mixture On CD4~+T And CD8~+T Cells Of Mycoplasma Pneumoniae Pneumonia Model Rats

Objective:To explore the effect of Zhiyan mixture on the expression of CD4~+T,CD8~+T and CD4~+T/CD8~+T in rats peripheral blood,and to explore the mechanism of Zhiyan mixture in the treatment of Mycoplasma pneumoniae pneumonia from the perspective of immunology.Methods:Forty-eight SPF Wistar rats,male and female,with body weight of200±20g,were randomly divided into blank group,model group,azithromycin group and Zhiyan mixture group,with 12 rats in each group.Modeling:In addition to the blank group,The other three groups were given 100μl,1×10~7CCU/ml MP bacterial liquid nasal drops for 3 consecutive days;The blank group was given the same amount of normal saline nasal drip for 3 consecutive days.Drug administration:After successful modeling,blank group and model group were given 2ml normal saline intragastrically,once a day;The azithromycin group was given Azithromycin Dispersible Tablets for 5 days,50mg/kg on the 1st day,25mg/kg on the following 4 days,and the same amount of normal saline was given intragastrically 5 days later,once a day.Zhiyan Mixture group was given 1ml Zhiyan Mixture by intragastric gavage,twice a day.Samples were collected in batches on the5th and 10th day after administration.Six rats were randomly selected from each group at a time.Peripherals of the rats were collected and the expressions of CD3~+T,CD4~+T,CD8~+T,CD4~+T/CD8~+T were detected.Lung and spleen were weighed and lung index and spleen index were calculated.The lung tissues were stained with HE to observe the histopathological changes.Results:1.HE staining:Compared with the blank group,the Zhi Yan Mixture group and the azithromycin group showed inflammatory injury;compared with the model group,the model group showed obvious inflammatory infiltration,and the two groups treated with medication were less severe than the control group.2.Lung index and spleen index results:on the 5th day,there was no significant difference between Zhiyan mixture group and azithromycin group(P>0.05),but there was significant difference among other groups(P<0.01);the change of lung index on the 10th day was the same as that on the5th day.There was no significant difference in lung index between groups on the 10th day and the 5th day(P>0.05).Spleen index:there was no significant difference between Zhiyan mixture group and azithromycin group on the 5th and 10th day(P>0.05),but there was significant difference among other groups(P<0.01).There was no difference between the groups on the 10th day and the 5th day(P>0.05).3.Flow cytometry results:Comparison of each group:on the 5th day,compared with the blank group,the expression of CD3~+T,CD4~+T and CD4~+T/CD8~+T in the remaining three groups decreased(P<0.05);Compared with the model group,the expressions of CD3~+T,CD4~+T and CD4~+T/CD8~+T in the remaining three groups were higher than those in the model group(P<0.05).CD8~+T was higher in model group than in the other three groups(P<0.05).There were no significant differences in the expression of CD3~+T,CD4~+T,CD4~+T/CD8~+T between azithromycin group and Zhiyan mixture group(P>0.05).At the 10th day of administration,there were no significant differences in the expression of CD3~+T,CD4~+T,CD8~+T and CD4~+T/CD8~+T in the remaining three groups except the model group(P>0.05).Comparison between groups:On day 10 and day 5,CD3~+T,CD4~+T,CD4~+T/CD8~+T in model group continued to decrease,but there was no difference(P>0.05),and CD8~+T had no significant change(P>0.05);The expression of CD3~+T and CD4~+T in Zhiyan Mixture group increased significantly(P<0.05),the ratio of CD4~+T/CD8~+T increased but had no difference(P>0.05),and CD8~+T had no significant change(P>0.05).In azithromycin group,CD3~+T increased to normal(P<0.05),and CD4~+T,CD8~+T,CD4~+T/CD8~+T increased slightly(P>0.05).Conclusion:1.Zhiyan Mixture can improve the pulmonary inflammation and general condition of MP-infected rats.2.Infection with MP can cause lung index to rise and spleen index to decline.Zhi Yan Mixer can affect lung index and spleen index,reduce lung tissue damage to a certain extent,and regulate immune disorders.3.The expression of CD3~+T,CD4~+T and CD4~+T/CD8~+T in the rat model of Mycoplasma pneumoniae pneumonia decreased,while the expression of CD8~+T increased.4.Zhiyan mixture can up regulate the expression of CD3~+T,CD4~+T and CD4~+T/CD8~+T in peripheral blood of rats with Mycoplasma pneumoniae pneumonia.The mechanism of Zhiyan mixture in Treating Mycoplasma pneumoniae pneumonia may be related to the regulation of immune mechanism in rats.