Exploration On The Mechanism Of Trametes Robinioplila In Adjusting The Imbalance Of Immune System In Asthmatic Children During Remission Stage

Objective:To evaluate the effect of Trametes Robinioplila on asthmatic children during remission stage and the mechanism of Trametes Robinioplila,we also explored its regulating effects on the T lymphocyte imbalance through the experience in vitro, in order to find the theoretical basis of the fungus for treating asthma in remission stage.Methods:Partâ… :The effect evaluation of Trametes Robinioplila on asthmatic children during remission stage.In clinical study,60 asthmatic children were divided into two groups,which were the Huaier group and the control group.All the children were followed up clinically being observed and the information was recorded as shown below:â‘ the sample number of one of the TCM symptoms before and after the drug administering;â‘¡the asthma control levels on patients evaluated by the questionnaire of ACT/C-ACT every 4 weeks at the beginnng of the first-care.Partâ…¡:The investigation of Trametes Robinioplila on the T lymphocyte in vitro in asthmatic children during remission stage.Peripheral venous blood(3ml) was collected from each of these children. PBMCs were isolated by density gradient centrifugation and seperated into three groups(the control group,the middle-dose group and the large-dose group).The samples were then incubated in vitro for 48 hours in the absence(the control group) or presence(the middle-dose group and the large-dose group) of Trametes Robinioplila in different concentration(0,3,6μg/ml).The expression of IFN-γ,IL-4,IL-10,T-bet, GATA-3 and Foxp3 in cellular precipitation was measured at mRNA levelby real-time fluorescence quantitative polymerase chain reaction technology(Real-Time PCR), and that of IFN-γ,IL-4 and IL-10 in supernatant was measured by ELISA. Statistical analysis:All Data were analyzed with SPSS 16.0.The data were numeration data in the partâ… ,and those were measurement data presented as Means±SD in partâ…¡.The P values are two-tailed and a P value of less than 0.05 is considered to be statistically significant. Results:Partâ… :1.The expression of TCM symptoms1.1 In the both groups,after the drug administering the pale complexion, spontaneous sweat,fatigue,white coating tongue,cersicolor exfoliative tongue and abnormal pulse presentation(thready and weak) got better than those before that (23vs14/21vs12,21vs11/20vs11,18vs9/15vs6,et al,P value＜0.05);especially,the spontaneous sweat in the Huaier group,cersicolor exfoliative tongue and abnormal pulse presentation in both groups improved markedly after the drug administering (21vs11,10vs1,12vs0,et al,p value＜0.01).But the differences of poor appetite showed no statistical significance in the same group(12vs6,10vs4,P value＞0.05).1.2 In the control group,after the drug administering the constipation and night sweat got no better than those before that(13vs8,22vs15,P value＞0.05),but in Huaier group,the symptoms improved markedly(12vs2,19vs7,P value＜0.01).1.3 Before the drug administering,the constipation and night sweat presented no significant differences between the Huaier group and the control group(12vs13, 19vs22,P value＞0.05);but after that,the two symptoms got better in the Huaier group than those in the control group(2vs8,7vs15,P value＜0.05).2.The expression of ACT/C-ACT2.1 Between before and after the drug administering,the differences of the asthma control levels presented no statistical significance(Pvalue＞0.05) in Huaier group.However at the 4^th and 8^th week in the control group,the differences of the asthma control levels were no better than the levels at the first(P value＜0.05).2.2 At the 4^th and 12^th week after the drug administering,the expression of the asthma control levels were insignificantly different between the Huaier group and the control group(P value＞0.05);but at the 8^th they were significantly different in the both groups(P value＜0.05).Partâ…¡:1.The expression of T LC cytokines at mRNA levelThe expression of the IFN-γmRNA was insignificant statistically among any two groups(2.21±0.26vs2.17±0.33vs2.10±0.31,P value＞0.05).The levels of the IL-4 mRNA in the large-dosel group were lower than those of the control group and the middle-dose group(0.86±0.13vs 1.28±0.14,0.86±0.13vs 1.21±0.17,p value＜0.01, P value＜0.05).The expression of the IL-10 mRNA in the large-dose group was higher than those in the control group(2.99±0.33vs2.63±0.36,P value＜0.05).2.The expression of T LC transcription factors at mRNA levelThe expression of the T-bet,Foxp3 mRNA in the large-dose group was lower than those in the control group(2.40±0.30vs2.63±0.25,2.31±0.31vs2.60±0.39,P value＜0.05).The levels of GATA-3 mRNA were markedly statistical significance between the large-dose group and the control group(1.56±0.22vs1.87±0.29,P value＜0.01),and there were statistical significance between the large-dose group and the middle-dose group(1.56±0.22vs1.71±0.24,P value＜0.05).3.The expression of T LC cytokines at protein levelThe levels of the IFN-γ,IL-10 in large-dose group were higher than those in the control group(672.78±184.72vs503.74±189.78,586.47±191.83vs431.67±135.25,P value＜0.05),but the differences of the IL-4 presented no statistic significance among any two groups(22.76±7.08vs21.17±6.84 vs18.73±4.55,Pvalue＞0.05).4.The ratio expression of the TLC cytokines＆transcription factorsThe levels of the IFN-γ/IL-4 mRNA in large-dose group were higher markedly than those in the control group,and there were significant differences between large-dose group and middle-dose group(2.61±0.59vs 1.91±0.61,2.61±0.59 vs2.00±0.62,P value＜0.01).But the differences of the T-bet /GATA-3 mRNA presented no statistic significance among any two groups(1.43±0.22vs1.52±0.22vs 1.55±0.24,P value＞0.05).The expression of the IFN-γ/IL-4 showed the statistic significance between large-dose group and control group(37.90±13.59vs 25.62±15.28, Pvalue＜0.05).Conclusions:1.Compared with the control group,the constipation and night sweat improved in the Huaier group;and at the 8^th week after the drug administering,the asthma control levels in Huaier group were better than those in the control group.It's demonstrated that Trametes Robinioplila could be benefit for the asthmatic children, improving the kids' body-constitution and adjusting the deficiency of that,in order to maintain the patient's condition.2.The experiences in vitro showed that Trametes Robinioplila could increase the levels of IFN-γin supernatant,and decrease the levels of IL-4 mRNA in cellular precipitation.The results indicated that Trametes Robinioplila could correct the drift of Th1/Th2 cytokines and regulate the imbalance of Th1/Th2 by up-regulating the Th1 cytokines in supernatant and down-regulating the Th2 cytokines. 3.The experiences in vitro presented that Trametes Robinioplila could down-regulate the expressions of GATA-3 mRNA in cellular precipitation,and the levels of T-bet mRNA were consumed.It's supposed that Trametes Robinioplila could inhibit the differentiation of Th0 into Th2 and adjust the ratio of Th1/Th2 cells by down-regulating the Th2 transcription factor.4.The experiences in vitro showed that Trametes Robinioplila could increase the expressions of IL-10 mRNA in cellular precipitation and IL-10 in supernatant,and the levels of Foxp3 mRNA were consumed.The results demonstrated that Trametes Robinioplila could alleviate the chronic allergic inflammation by up-regulating anti-inflammatory cytokine;meanwhile the drug could induce the differentiation of ThO into CD4⁺CD25⁺ Treg,in order to inhibit the Th2 and adjust the imbalance of Th1/Th2 cells.