| Objective:1. To establish the rat models of acute allergic asthma with the intestional microflora disruption.2. To explore the relationship of alteration of gut microecology and the immune regulatory of lung and intestine.3. To cure model rats from three aspects to compare the validity between them:the treatment to lung and gut, the treatment to lung and the treatment to gut. To confirm the relationship of gut microecology and the immune regulatory of lung and intestine and seek evidence for the traditional theory of "Lung and Gut are interiorly-exteriorly related".Methods:1. SD mice were given oral cefoperazone and inoculated with Candida albicans by oral to establish the disruption of intestional microflora, then to establish the allgic asthma model by injection and atomization of OVA. To test the validity of the model rat from the individual level, the cell level and the molecule level.2. To explore the immune relationship between gut and lung by analyzing 16SrDNA of intestinal microflora and the mRNA expression levels of TLR2, TLR4> MyD88、TARF6 and β-arrestin in lung and gut tissues.3. To confirm the immune relationship between lung and gut and compare the validity of different three therapeutic methods by using "ShenJiangSan" "GuiZhijiaHouPuXingZi decoction" and "ZengYeCengQi decoction" to cruel the model rats. To seek evidence for the traditional theory of "Lung and Gut are interiorly-exteriorly related".Results:1. Evaluation on the effectiveness of the model rat.In the model rat, the respiration rate,the number of eosinophils in BALF and blood,the number of lymphocyte,neutrophile granulocyte and mononuclear cells in blood and the level of IgE were significantly higher than the control group. The level of slgA were lower than the control group. All the difference between two groups had statistical significance. The number of IL-4 and IFN-y were obviously been changed in the model group.The GATA-3 mRNA was higher in the model group than the control group which illustrate imbalance of Thl/Th2.2. The mechanism of the immune relationship between lung and intestinal microflora.In comparison with the normal group and the positive group,the diversity of intestinal microflora had been obviously changed in the model group,including the increase of harmful bacteria like Clostridia and Helicobacter and the decrease of beneficial bacteria like Bacteroidetes and Proteobacteria. The mRNA expression levels of TLR、MyD88、TRAF6 and P-arrestin of lung in model group which vary in accordance with each other were significantly higher than the other groups. The mRNA expression levels of TLR4、MyD88、TRAF6 and β-arrestin of gut tissues in model group were higher than the other groups, significantly in TLR4 mRNA.The varies of the mRNA expression levels of MyD88 was in accordance with TRAF6 and P-arrestin.3. The comparison of the effect to the model rat of three different traditional Chinese medicine prescription."jiaweiShengJiaSan"had the best effect in the recovery of respiration rate,hyperresponsiveness, pathological section of lung, the number of eosinophils in BALF and the level of IgE, T-bet/GATA-3,IFN-y and IL-4,while also had effect in recovery of the diversity of intestinal microflora but inferior to the group of treatment to gut; "ZengYeCengQi decoction" had the best function in the recovery of intestinal microflora which also had the good mediation of hyperresponsiveness, the number of white blood cell in BALF and the number of eosinophils and neutrophile granulocyte in blood. All these differences between two groups had statistical significance. "GuiZhijiaHouPuXingZi decoction" had advantage in the regulation of respiration rate, the number of eosinophils in blood and the level of T-bet, but inferior to the group of treatment to gut on the recovery of intestinal microflora. The mRNA expression levels of TLR2, MyD88 and TRAF6 in lung tissues of treatment groups were significantly lower than the model. Only in the group of treatment to lung and gut and the group of treatment to lung had statistical significance comparing with model groups in the mRNA expression levels of TLR4 and β-arrestin. The mRNA expression levels of TLR2,MyD88 and TRAF6 in gut tissues of treatment groups were all higher than the model,but the difference of TLR2 with the model was significant in the group of treatment to lung and gut and the group of treatment to lung,the difference of TRAF6 with the model being significant in the group of treatment to lung and treatment to gut. The mRNA expression levels of β-arrestin in the group of treatment to gut was significantly higher than the model.Conclusions:1. Estabilish the rat models of acute allergic asthma with the intestional microflora disruption by given oral cefoperazone and Candida albicans, injected and atomized OVA.2. The immune regulation of lung and gut all could be regulated by TLRs/NF-κB signaling pathway in the rat of acute allergic asthma with the intestional microflora disruption,on which β-arrestin had promoting effect. The TLRs/NF-κB signaling pathway of lung tissues had the contrary regulation to Thl/Th2.3. All the treatment groups had effect both on intestional microflora and pathologic indexes of allergic asthma which indicated all the treatments could regulate both lung and gut on different levels. The treatment to lung and gut group had the best effectiveness. The TLRs/NF-κB signaling pathway including MyD88 played important role both in lung and gut in all treatment groups. The up-regulation of signaling pathway occurred in lung tissues, down-regulation in gut tissues. The differences between three treatment groups confirmed the traditional prescription of Lung and Large Intestine are interiorly-exteriorly related", and revealed the treatment to lung and gut had the best effect in the recovery of acute allergic asthma with the intestional microflora disruption. |
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