The Study Of Neuronal Acetylcholine Receptor Alpha 7 Expressed In Peripheral Blood CD₄～+T Lymphocytes Of Asthma Children

ObjectivesTo analysis the relationship between nAChRα7 expressed in peripheral blood CD4~+ T lymphocytes of asthma children and cytokine IFN-γ,IL-4 in lymphocyte cell culture fluid, to explore the relationship between express of nAChRα7 and Th1/Th2 balance, to explore the clinical significance of nAChRα7 in the pathogenesis of asthma.Materials and MethodsAccording to the diagnostic criteria routine of prevention and cure of asthma,30(male 16,female 14) onset typical asthma children were selected and 20(male 11,female 9) health as contrasts. Get 8ml empty stomach venous blood, anticoagulated with EDTA, equal number lymphocytes ( 2×10~6 ) were separated and cultured in equal volume culture medium (3ml) , with blank, nicotine(100uM),α-bungarotoxin (1ug/ml}respectively. After 24 hours, culture fluid were collected and conserved under -20℃, lymphocytes were detected under flow cytometry. CD3,CD4,nAChRα7 were assayed. Using ELISA detect IFN-γ,IL-4 in culture fluid.Results1.Consequence of flow cytometry:①nAChRα7 is expressed both in normal and asthma children. Enhanced nAChRα7 expressed in peripheral blood CD4~+ T lymphocytes of asthma children than health contrasts (3.91±2.67%/2.90±1.01 %, t=2.17, P<0.05) (3.91±2.67% /2.90±1.01%,t =2.17, P<0.05).②Enhanced expressed of nAChRα7 in peripheral blood CD4~+ T lymphocytes after stimulated with nicotine (control group 4.86±2.82%/2.90±1.01 % ,t=3.03 ,P<0.01,asthma group 6.73±2.49%/3.91±2.67%, t=4.17, P<0.01), and enhanced express in asthma group than contrast group after stimulate with nicotine.(6.73±2.49%/4.86±2.82% t=2.35, P<0.05 )③Slightly decrease after stimulated withα-bungarotoxin, no statistically significant (control group2.40±3.18%/2.90±1.01 %, t=0.86, P>0.05, asthma group 3.23±3.65%/3.91±2.67%, t=1.01, P>0.05) .2.Consequence of ELISA assay:①Lymphocyte culture fluid of asthma children compared with health children: IL-4 increased (6.35±1.62/4.69±0.79 pg/ml, t=4.74, p<0.01), IFN-γreduced (2.04±0.94/2.84±1.17 pg/ml, t=2.50, p<0.05);②Incubation with nicotine: IL-4 increased (control group 5.87±1.12/4.69±0.79pg/ml, t=3.76,P<0.01,asthma group 7.64±1.51/6.35±1.62pg/ml, t=3.14, P<0.01) ,IFN-γreduced (control group 1.91±0.72/2.84±1.17pg/ml,t=3.44, P<0.01,asthma group 1.44±0.64/2.04±0.94pg/ml, t=2.86,P<0.05 );and IL-4 enhanced,IFN-γreduced obviously in asthma group compare with control group.③Incubation withα-BTX: IL-4 reduced (control group 4.26±1.69/4.69±0.79pg/ml, t=1.05, P>0.05,asthma group 5.80±1.20/6.35±1.62pg/ml, t=1.27, P>0.05) ,IFN-γ(control group 3.37±0.94/2.84±1.17pg/ml, t=1.36, P>0.05,asthma group 2.18±1.38/2.04±0.94pg/ml, t=0.45, P>0.05).3. After incubation with nicotine 24 hours, the up-regulation of nAChRa7 in CD4~+T lymphocyte and the increase of IL-4 present positive correlativity (r=0.517, p<0.01),decrease of IFN-γpresent negative correlativity (r=-0.288, p<0.05).Conclusions1. Asthma children exist immune unbalance, IL-4 enhanced, IFN-γreduced;.2. There are up-regulation of nAChRa7 in CD4+T lymphocyte in asthma children . The nAChRα7 of CD4+T lymphocyte present positive correlativity with IL-4 ,and present negative correlativity with IFN-γ.