| Objective:To observe the effect of kangzhi syrup on the TLR4 and Thl/Th2 related cytokines in mycoplasma pneumoniae infection by this experiment,discussthe regulation effect of kangzhi syrup on its immune mechanism,provide the basis for a further study and clinical application.Methods:1 20 wistar male rats weredivided into 4 groups randomly:there are blank control group,model control group,kanghzi syrup group andazithromycin group.There were 30ratsin each group.The blank control group and the inoculation group were kept in separate rooms,and the experiment began after 1 week of adaptive feeding.Infection methods:in addition to the blank group,the remaining groups were inoculated 106CCU/ml of MP for 4 consecutive days.The blank control group was given an equal amount of medium without MP.The drug intervention method:after the final infection of each group,the drug was administered,and the kangzhi syrup was given 6g/kg of weight,and the volume was set to 1 ml/100g.Azithromycin was administered on the 5 th day,with azithromycin 45mg/kg on the first day,and azithromycin 22.5mg/kg on theother four days.The blank control group and the model control group were given saline irrigation respectively.Each group was administered one time a day for 10 consecutive days.Each group respectively in experimental animals for the first time to give medicine after 1d,3d,5d,8d,10d(this period was defined as the period of observation),each group randomly take 6 rats,through HE,ELISA and Western blot and Real-time PCR technology,from histological level,cellular level and molecular level respectively to observe the resistance syrup on animal model of lung pathological changes,cytokines,inflammatory mediators and related factors of the influence of protein and gene expression.Results:1.The detection of MP in BALF:The 1st day of the observation,the model control group can detect MP in BALF;Compared with the model control group,there was a significant decrease of MP in kangzhi syrup group and the azithromycin group in the 3,5,8 and 10 days,with statistically significant difference(P<0.05).Compared with azithromycin group,no significant difference was observed during the whole observation period in the kangzhi syrup group(P>0.05).2.HE staining:The model control group showed aresponse of inflammatory on the first day of the observation period,the third day was the most severe,and the wall of bronchial tube was damaged,and the mucosal hyperemia and edema were observed.There are more inflammatory cells around the bronchus,mucous membranes and blood vessels,some of the alveolar walls are thinner or ruptured,and the alveolar space is widened,and the surrounding area is surrounded by inflammatory cells.Compared with the control group,the inflammatory response of the kangzhi syrup group and azithromycin group was significantly reduced.3.Lung histopathological score:The model control group showed an inflammatory response during the first day of the observation period,and the pathological score to a peak at the 3rd day.Compared with the control group of the model,the pathological scores of the two groups were significantly decreased in the 3,5,8 and 10 days of the observation period,and the differences were statistically significant(P<0.05).Compared with the azithromycin group,there was no significant difference in pathology score during the whole observation period in the kangzhi syrup group(P>0.05).4.ELISA test showed:4.1 ELIS A testofserum results showed that the content of TLR4,MyD88 and NF-?B in serum of the control group was significantly increased compared with that of the blank control group(P<0.05).Compared with the control group of the model,the contents of TLR4,MyD88 and NF-?B were significantly reduced in the 3,5,8,10 days of the observation period in the two treatment group(P<0.05).At the same time,there was no significant difference in TLR4,MyD88,and NF-?B in the whole observation period(P>0.05).4.2ELISA test of BALF showed that compared with the blank control group,the content of IL-6 and TNF-? in BALF in the model control group was significantly increased,and the content of IL-2 in BALF was significantly decreased(P<0.05).Compared with the model control group,the contents of IL-6 and TNF-?were significantly reduced in the 3,5,8,10 days of observation period,and the levels of IL-6 and TNF-?were significantly decreased in BALF of the two treatment group(P<0.05).At the same time,there was no significant difference in the content of IL-6,TNF-?and IL-2 during the whole observation period(P>0.05).5.Western Blot test results showed that compared with the blank control group,the expression of IFN-?protein expression in the lung tissue of the control group was significantly reduced,and the expression of IL-4 protein was significantly increased(P<0.05).Compared with the modle control group,the expression of IFN-? protein expression was significantly increased in the 3,5,8,10 days of observation period,and the expression of IL-4 protein was significantly decreased(P<0.05).There was no significant difference(P>0.05)during the whole observation period between the kangzhi syrup group and the azithromycin group.6.Real-time PCR test results showed that,compared with the blank control group,the expression of IFN-?mRNA in the lung tissues of the model control group was significantly reduced,and the expression of IL-4mRNA was significantly increased(P<0.05).Compared with the model group,there was a significant increase in the expression of IFN-?mRNA in the two treatment group,and the expression of IL-4mRNA was significantly decreased in the 3,5,8,and 10 days of observation period(P<0.05).There was no significant difference(P>0.05)during the whole observation period between the kangzhi syrup group and the azithromycin group.Conclusion:1.Kangzhi syrup can effectively improve the general condition of MP infected rats,and has a significant inhibitory effect on the inflammatory response of lung tissue.2.Kangzhi syrup can reduce resistance MP infection in rats serum content of TLR4,MyD88,the NF-?B,we hypothesized that TLR4-NF-?B can be used as evaluation of mycoplasma infection of an ideal,weakened the TLR4-NF-?B transduction drug is expected to become a new targets for the treatment of MPP.3.Kangzhi syrup can reduce the content of IL-6 and TNF-ain the rats with MP infection,and increase the content of IL-2 in the BALF of rats with MP infection,thereby inhibiting the inflammatory response of MPP.4.Kangzhi syrup can raise IFN-y mRNA and protein expression,cut IL-4mRNA and protein expression,and regulate the Thl/Th2 balance of the body,thereby inhibiting inflammation of MPP,protect the lung tissue.The mechanism of the treatment of mycoplasma pneumoniae pneumoniae by kangzhi syrup may be related to the regulation of immune balance. |
PDF Full Text Request