Effects Of Qing-fei-pei-yuan Granules On IL-17、JAK-STAT Signaling Pathway Of Pulmonary Infection In Immunocompromised BALB/c Mouse Model

Objective: The objective of this paper is to investigate the effects of theQing-Fei-Pei-Yuan granules on the secretion of IL-17cytokines of serums and theactivation of JAK2-STAT3signal transduction pathway of lung tissues thoughmodeling immunocompromised pulmonary infected BALB/c mice model as well asillustrate the partial action mechanisms of Qing-Fei-Pei-Yuan granules onimmunocompromised pulmonary infection. It will provide the experimental basis forthe further researches and the clinical applications of Qing-Fei-Pei-Yuan granules.Methods：(1) Modeling and administration: The60clean and healthy BALB/c femalemice were adaptively feeding3days. Randomly selected12mice as Normal Groups.The remaining mice will be injected FLV virus into their abdominal cavities. Thenafter21days these mice will be instilled streptococcus pneumoniae to make the modelof immunocompromised pulmonary infected mice. After successful modeling, themodeling mice were randomly divided into four groups, which areImmunocompromised Pulmonary Infected Group (Model Group), Zidovudine Group,Tang Herb Group, and Qing-Fei-Pei-Yuan Group. Each treatment group will be giventhe corresponding drugs. Normal Group and Model Group were given a gavage ofsame volume of normal saline in14days.(2) Indication Measuring: Observingexperimental mice and recording their state.6hours after the last administration, theirserum and lung tissues will be taken and cryopreserved in-80℃environment. Thechange of IL-17cytokines of each group will be examined by enzyme-linkedimmunoassay (ELISA); The secretion of JAK2and p-STAT3of each group will beexamined by Western Blot; The secretion of JAK2and STAT3mRNA of each group will be examined by Real time PCR.(3) Statistical Process: Using SPSS17.0softwareanalyzing the results of IL-17, JAK2, and p-STAT3， P<0.05is the difference withstatistical significance. The differences of the results of JAK2and STAT3mRNAbetween each group will be calculated based on2-ΔΔCtMethod. The results aremeaningful when2-ΔΔCt>2or2-ΔΔCt<0.5.Results：(1) The general change of mice: Immunocompromised pulmonary infectedBALB/c mice modeling successfully, all of Qing-Fei-Pei-Yuan group, Zidovudinegroup, and Tang Herb group could improve the general state of modeled mice, such asthe growth of mice’s weight, and the mitigation of dyspnea et al. especiallyQing-Fei-Pei-Yuan group which could prolong latent phase of coughing andgasp for breath, mitigate abdominal muscle twitching, vivify state, and ameliorate furcolor. The results of Zidovudine and Tang Herb groups come second.(2) The resultsof IL-17cytokines of mice’s serums: The secretion result of IL-17of the mice inModel Group increased comparing with Normal Group (P<0.05). Such result ofQing-Fei-Pei-Yuan Group is less than Model Group (P<0.05). The differences of theresults between Zidovudine, Tang Herb, and Model Groups do not have statisticmeaning comparing with Model Group (P>0.05).(3) The secretion results of JAK2ofmice’s lung tissues: The secretion results of JAK2of the mice in Model Group ishigher than Normal Group (P<0.05); Comparing with Model Group, the results inother groups are all raised (P<0.05). Also comparing the results between each group,The secretion results of JAK2of Qing-Fei-Pei-Yuan Group is less than Zidovudine,and Tang Herb groups (P<0.05). The differences of the results between Zidovudineand Tang Herb Groups do not have statistic meaning (P>0.05).(4) The secretionresults of JAK2mRNA of mice’s lung tissues: the JAK2mRNA of the mice in ModelGroup is higher than Normal Group (2-ΔΔCt>2); Comparing with Model Group, JAK2mRNA of the mice in the other groups are all reduced (2-ΔΔCt<0.5). The secretionresults of JAK2mRNA of Qing-Fei-Pei-Yuan Group reduced comparing withZidovudine Group with meaningful result (2-ΔΔCt＜0.5). Comparing with Tang HerbGroup, the differences of such result of Qing-Fei-Pei-Yuan Group is unmeaningful(0.5＜2-ΔΔCt＜2).(5) The secretion results of p-STAT3of mice’s lung tissues: Thesecretion results of p-STAT3of Model Group is higher than Normal Group (P<0.05);Comparing with Model Group, such results of Qing-Fei-Pei-Yuan and Tang Herbgroups reduced (P<0.05) while Zidovudine Group raised (P<0.05). Also the result ofQing-Fei-Pei-Yuan Group is less than Tang Herb and Zidovudine groups with statistic meaning (P<0.05).(6) The secretion results of STAT3mRNA of mice’s lung tissues:The secretion result of STAT3mRNA of Model Group is higher than Normal Group(2-ΔΔCt＜0.5); Comparing with Model Group, the results of Qing-Fei-Pei-Yuan andZidovudine groups reduced (2-ΔΔCt＜0.5) while Tang Herb with unmeaningful result(0.5＜2-ΔΔCt＜2). Also the result of Qing-Fei-Pei-Yuan Group is less than Tang HerbGroup (2-ΔΔCt＜0.5) while unmeaningful comparing with Zidovudine Group (0.5＜2-ΔΔCt＜2).Conclusion：(1) Qing-Fei-Pei-Yuan granules could improve the general state ofmodeled mice. Its effectiveness was proved through active indication of mice.(2) Theincreasing secretion of IL-17and the active signal transduction pathway ofJAK2-STAT3are related with the immunocompromised pulmonary infection.(3)Qing-Fei-Pei-Yuan granules could efficiently inhibit on the secretion of IL-17cytokines of mice’s serums. It shows that the mechanism of anti-inflammatory actionof Qing-Fei-Pei-Yuan granules is related with the secretion of IL-17.(4)Qing-Fei-Pei-Yuan granules could efficiently inhibit the activation of JAK2-STAT3signal transduction pathway, reduce the transcriptional expression of JAK2, interruptactivation of JAK2and STAT3, and inhibit the generation of p-STAT3. This showsthat the mechanism of immunity enhanced and anti-infective action ofQing-Fei-Pei-Yuan granules is related with the inhibition of the activation ofJAK2-STAT3signal transduction pathway.(5) Qing-Fei-Pei-Yuan granules couldcontrol the secretion of IL-17and inflammation, improve the state ofimmunocompromised pulmonary infection through inhibiting the activation ofJAK2-STAT3signal transduction pathway.