| Objective:1. Establishment of asthma rat model with kidney qi deficiency.2. To explain the mechanisms of asthma rat model with kidney qi deficiency and the therapeutic effect of Yishenchuanning decotion(YSCN) in the whole viewing from immunology and molecular biology, in order to offer more advantageous experimental evidence for clinical application.Methods:1. Based on TCM etiology, establish asthma with kidney qi deficiency rat model applied methods of multi-factors combined intraperitineal sensitized liquid and breathing atomizated egg albumin consecutively.2. The male Wistar rats were randomly divided into normal control group, asthma group, kidney qi deficiency asthma group, Western medicine group and Yishenchuanning decotion (YSCN) group, each group had 10 rats. After the intervention for 14 days continuously, the drug groups began to medicine. After that, the enzyme linked immunosorbent assay (ELISA) was used to test the expression of IL-6、IL-10、IL-17、TGF-β1 in BALF. Moreover, effects of YSCN on asthma kidney qi deficiency syndrome of inflammatory cytokines to be further investigated.3. The expression of Foxp3、RORγt、TGF-β、IL-6 in the lung tissue were detected by Western blotting. The therapeutic machanisms of YSCN on asthma with kidney qi deficiency were investigated according to the effect of this preparation on immune markers.Results:1. Model rats with asthma of kidney qi deficiency syndrome were set up successfully.2. Compared with the normal control group, IL-6、IL-17 concentration increased, while IL-10、TGF-β1 decreased in asthma group and kidney qi deficiency asthma group(p<0.05). Between the asthma group and the kidney qi deficiency asthma group, the change of IL-6、 IL-17、TGF-β1 in BALF were significantly(p<0.05). Compared with the kidney qi deficiency asthma group, IL-6、IL-17 concentration decreased, while IL-10、TGF-β1 increased in drug groups(p<0.05). Compared with the Western medicine group, IL-17、IL-10 changed significantly in Yishenchuanning decotion (YSCN) group(p<0.05).3. Compared with the normal control group, lung tissue TGF-β、RORγt、IL-6 protein expression increased and Foxp3 protein expression decreased(p<0.05). Between the asthma group and the kidney qi deficiency asthma group, the change of Foxp3 protein expression were significantly(p<0.05). Compared with the kidney qi deficiency asthma group, TGF-β、 RORγt、IL-6 protein expression decreased and Foxp3 protein expression increased in drug groups(p<0.05). Compared with the Western medicine group, TGF-β、 RORγt、Foxp3 protein expression changed significantly in Yishenchuanning decotion (YSCN) group(p<0.05).Conclusions:1. Multi-factors combined egg albumin applied in the research to establish asthma with kidney qi deficiency rat model were proved to be stable and reliable.2. In kidney qi deficiency asthma group, the unbalanced Th17/Treg contributes to both high frequencies of IL-6、IL-17 and low frequencies of IL-10、TGF-β1, and the abnormal expression of protein levels of TGF-β、RORγt、IL-6、Foxp3, the ratios may evaluate the immune disbalance of kidney qi deficiency asthma objectively. Imbalance of Treg and Th17 may play a key role in the formation of.the Kidney Qi Deficiency syndrome Asthma. Inflammatory reactions were aggravated by kidney qi deficiency in airway and lung tissue, which is through regulating cytokines and transcription factors expression.3. Yishenchuanning decotion can adjust the imbalance Th17 and Treg, and related the expression of cytokines and transcription factors, thus improving the state of immune disorders in kidney qi deficiency syndrome asthma rats, thereby restoring immune kidney qi deficiency syndrome asthma rats tolerated.4. The immunological characteristics and their machanisms of asthma rat model with kidney qi deficiency are interpreted integrally. The therapeutic mechanisms of YSCN on the asthma are displayed systematically, showing the characteristics of YSCN Multi-channels, multi-targets comprehensive therapy. |
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