| ObjectiveTo investigate the curative effect of Dasangju mixture(DSJ) on macrolide-resistant myeoplasma pneumoniae pneumonia(MRMPP) in children and its mechanism on changing the cytokines. To study the impact of DSJ on mouse RAW 264.7 inflammatory cell model and the signaling pathway of nuclear factor kappa B(NF-k B), which could be used to investigate the mechanism of anti-inflammatory immunity and provide the clinical evidence and experimental basis for treating pediatric myeoplasma pneumoniae pneumonia.MethodsDesigned as a prospective randomized control test,60 patients with MRMPP in children were randomly divided into two groups, which were named as treatment group and control group, and there were 30 cases in each group. Patients in the control group were treated with the azithromycin(AZM) as well as the basic symptomatic treatments. And children in the treatment group were treated with DSJ, associated with the medication used in the control group. The symptoms and signs were observed and the laboratory examination of MP-IgM, IL-6 and IL-10 were tested by ELISA. Moreover, drug resistance of myeoplasma pneumoniae was tested by etiological examination.Designed as a RAW 264.7 cell culture experiments in vitro, the effects on proliferation were tested by cell counting method. Then, established the inflammatory celluar model by lipopolysaccharide(LPS), RAW 264.7 cell would be treated with DSJ and AZM.3 hours after the intervention, TNF-a would be tested, while IL-6 and IL-10 would be tested after 6 hours. Finally, the expressions of NF-kB p65 were tested by Western blot.Results1. The total effective rate of treatment group was 86.67%, while it was just 56.67% in the control group, and the curative effect of treatment group was signicantly better than the control group(P<0.05). The alleviating time of the symptoms including fever,cough and the signs like lung rale in the treatment group was statistically less than the control groups (P<0.05). The difference of IL-6 and IL-10 serum concentrations before and after medication in the treatment group were remarkablely more than the control group (P<0.05). Moreover,2 cases with gastrointestinal adverse reaction were in the treatment group, but 11 cases were in the control group (P<0.05). And 9 patients with abnormal liver function in the control groups while there were just 2 cases in treatment group, since there was no significance(P>0.05).2. In celluar experiments on proliferation, the counts of low concentration group of Dasangju mixture (LDSJ), medium concenctration group of Dasangju mixture (MDSJ) and high concentration group of Dasangju mixture (HDSJ) were notly more than the blank group (P<0.05). However, the counts of AZM group were slightly less than the blank group, and there was no significance (P>0.05). Through making fitted curve for the LDSJ, MDSJ and HDSJ, there was positive correlation between each concentration groups on the cell proliferation(y= 0.0987 x+2.73, r= 0.99).3. In vitro experiments of the inflammatory model, the concentrations of TNF-a and IL-6 in HDSJ, MDSJ and LDSJ were remarkablely less than the control group (P<0.05). And the concentrations of TNF-a, IL-6 and IL-10 in each groups of DSJ and AZM as well as the control group were much more than the blank group(P<0.05). Indeed, the concentrations of TNF-α, IL-6 and IL-10 in HDSJ were notly less than the control group(P<0.05). And it was significantly less for the concentrations of TNF-α amd IL-6 in the AZM group than in the control group (P<0.05), while it was weakly less for the concentration of IL-10 in the AZM group than in the control group (P>0.05). Moreover, the concentrations of IL-6 in the MDSJ and LDSJ were as much as the AZM group, and there was no significance (P>0.05). In addition, the concentrations of IL-10 in the HDSJ and MDSJ were much less than the control group (P<0.05), but it was weakly less in the LDSJ than in the control group(P>0.05).4. In the research for the signaling pathway, the relative expressions of NF-k B p65 in each groups of DSJ and AZM were less than in the control group(P<0.05). Since the relative expression of NF-k B p65 in the LDSJ was as much as the control group and there was no significance (P>0.05), the relative exressions of NF-k B p65 in the MDSJ and HDSJ were remarkablely less than the contropl group (P<0.05). Moreover, there was no significance between the MDSJ and HDSJ. And the relative expressions of NF-k B p65 in each groups were much higher than the blank group (P<0.05).Conclusion1. DSJ could be used to reduce the time required to alleviate the symptoms and signs of patients with MRMPP, including antipyresis, relieving cough and reducing rales, which was significantly better than simply using western medicines for macrolide-resistant myeoplasma pneumoniae. Meanwhile, it could improve the efficacy through the mechanism of anti-inflammatory immunity to regulate the T helper cell and its cytokines of subsets.2. DSJ could promote cell growth and development, which may improve the mucous rehabilitation. Moreover, not only could DSJ reduce the expression of cytokines like TNF-a, IL-6, but also it could lead the expression of IL-10, a two-way regulation factor, to reduce to the relative concentration for anti-inflammation. And for dynamic equilibrium and anti-inflammatory immunity, NF-k B p65 was inhibitted by DSJ to induce the inflammatory cytokines secretion. |
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