The Effect Of Peroxisome Proliferator-activated Receptor-γ Agonists On IL-4/ STAT6 In T-Lymphocytes Of Patients With Asthma

Objective:To explore the effect of Peroxisome Proliferator-activatedReceptor-γ(PPAR-γ)Agonists on interleukin-4/Signal transducer andactivetor of transcription 6 (IL-4/STAT6) signaling pathway inT-lymphocytes of patients with asthma.Methods:The PBMCs were separated from 30mL peripheral blood of 10patients with asthma and 10mL peripheral from 10 healthy volunteers.The PBMCs were maintained at humidified atomosphere of 95%air/5%CO2 at 37℃for four hours in fresh RPMI-1640 mediumsupplemented with 10% FCS (fetal calf serum), 3 per ten-thousand ofglutamine, 100U/mL penicillin, 100 U/mL streptomycin and 50ng/mLPHA(complete RPMI-160 medium). The cell-suspension of PBMCs hadbeen cultured by complete RPMI-160 medium. Then T-lymphocytes werecollected from the PBMCs which have been deal with adherenceT-lymphocytes were divided into four groups as following (the density ofT-lymphocytes was 1×10~6 per milliliter, each well was two milliliter): ahealthy control group (group A), an asthma control group (group B); anasthma group being cultured with 2.5μg/mL IL-4RAb (C group), an asthma group being cultured with 100mmol/ml Rosiglitazone (D group),an asthma group being cultured with 2.5μg/mL IL-4RAb and100mmol/ml Rosiglitazone(E group); Among above groups, at frist, weadded IL-4RAb in group B and group E, Let IL-4RAb well combinedwith IL-4R which resulted IL-4 fail to bind IL-4R, one hour later, thenadding Rosiglitazone in group C and group E; Finally, each of groupswere cultured for 48 hours. The concentration of IL-4 in supematantliquid of each groups were detected by ELISA. The express ofphosphorylation of STAT6 in the T-lymphocytes was determined byWestern blot. The data were expressed with mean and standard deviation((?)±s). Statistic was analyzed with SPSS 12.0 software. Ifαis 0.05 anddouble P is less than or equal to 0.05, the results have achieved statisticalsignificance.Results:1 The results of ELISA(1) The concentration of IL-4 in the complete RPMI-1640 medium wasnot discovered.(2) The concentration of IL-4 was secreted by T-lymphocytes in groupA was higher than group B (P＜0.01).(3) The concentration of IL-4 in groups of patients with asthma wasgroup B＞group C＞group D＞group E (P＜0.01).2 The results of Western blot (1) The express of phosphorylation of STAT6 on T-Lymphocytes ingroup A was lower than group B (P＜0.01).(2) The express of phosphorylation of STAT6 in groups of patientswith asthma was group B＞group C＞group D＞group E (P＜0.01).3 Correlation of analysis:In group B, there was positive correlation between the level ofphosphorylation of STAT6 in T-lymphocytes and the concentration ofIL-4 in supematant liquid. (r=0.86, P＜0.01).Conclusion:1 The concentration of IL-4 was secreted by asthmas' T-lymphocytesand the level of phosphorylation of STAT6 in the asthmas'T-lymphocytes were higher than the healths' T-lymphocytes;2 IL-4 which was secreted by the T-lymphocytes of patients withasthma could increase the level of phosphorylation of STAT6;3 IL-4R Ab and.Rosiglitazon could decrease both the concentration ofIL-4 and the level of phosphorylation of STAT6 in patients withasthma;4 Rosiglitazon could decrease the express of phosphorylation of STAT6by means of decreasing the concentration of IL-4; moreover, it couldalso decreased the express of phosphorylation of STAT6 in otherways;...