Study On The Expression Of Related Molecules In Hypersensitivity Th17 Cells And The Intervention Mechanism Of Allergic

Objective:To investigate the micro-mechanism of allergic constitution-imbalance of Thl7 cell differentiation, Looking for effective specific molecular markers of allergic constitution, and the target of the TCM formula "Guo Min Kang" treating allergic constitution and curing allergic disease, from the perspective of TCM constitution theory, to further explore the pathogenesis and treatment of allergic disease, expecting to do guidance for its clinical treatment.Methods:Basing on the Classification and Determination of Constitution in TCM and the allergen test, we included in the 44 subjects, who were physical examinees from TCM constitution identification center of Beijing university of Chinese Medicine Guoyitang clinic, et al, who were required that were working or living in Beijing for a long time and aged between 18 to 40 years old. Among them, there were 20 ones in mild constitution group as control group, and 24 in allergic constitution group as experimental group, which should rule out ones with other disease diagnosing by modern medicine, women who were pregnant or in lactation or preparing for conception. Ensure that the baseline data of each group were comparable, and made sure that during the intervention experimental group won’t take other allergic drugs, and signed informed consent forms. Mild constitution, allergic constitution before and after 3 months’medical intervention was draw blood respectively, collect 10ml of blood on empty stomach in the early morning,36ul whole blood was pre-treated according to the manual of QuantiGene plex kit and the remaining blood was separated serum, the cryopreserved respectively in -80℃ degrees. The serum is for testing protein expression levels of IL-6, TGF-betal, IL-lbeta, IL-17A, IL-17F by protein chips; pretreated whole blood for detect gene level of STAT3, JAK2 mRNA by QuantiGene Plex. Data were analysed with spss20 software, differences between the mild constitution and allergic constitution with One-Way ANOVA, differences between allergic constitution before and after 3 months’medical intervention with paired t-test, the data were expressed as x±s, P<0.05 means statistical difference, P<0.01 stands for significantly statistical difference.Results:1. Allergic constitution group compared with mild constitution group, average levels of serum protein IL-6 of allergic constitution group are increased than peace constitution control group, the difference was not statistically significant (P>0.05); levels of TGF-betal, IL-17A, IL-17F protein are higher than the mild constitution control group, the difference was statistically significant (P<0.05); levels of IL-lbeta protein is higher than the mild constitution control group, difference has the obvious statistical significance (P<0.01); average levels of STAT3 and JAK2 mRNA of allergic constitution are higher than mild constitution control group, difference was not statistically significant (P>0.05).2. Allergic constitution compared before and after the intervention, the average level of serum IL-6, TGF-beat1, IL-lbeta, IL-17A after the intervention is lower than before, difference were not statistically significant (P>0.05); IL-17F levels is decreased than before the intervention, the difference was statistically significant (P<0.05); after the intervention average levels of STAT3 and JAK2 mRNA are decreased than before, the levels are almost the same as the control group, difference was not statistically significant (P>0.05).Conclusion:1. TGF-betal, IL-17A, IL-17F, IL-lbeta are the different factors between mild constitution and allergic constitution. Especially IL-lbeta is of significant difference between allergic constitution and mild constitution, which is visible as the potential auxiliary index for clinical judgement of the allergic constitution.2. Differences of Th17 cells differentiation between allergic constitution and mild constitution may not involve in the activation of JAK2/STAT3 pathway, specific pathways mechanism needs further research.3. IL-17F is the target of "Guo Min Kang" treating allergic constitution; the average levels of IL-17A, IL-6, TGF-betal, IL-lbeta are decreased after treatment than before, but the differences are not statistically different, maybe because formula using time is too short, we should further extend the duration, to do a further observation.