Stat6 Antisense Oligonucleotide On Asthma Study On The Effect Of Spleen Lymphocytes In Mice

Objective To explore the effect of STAT6 antisense oligonucleotid on spleen lymphocytes of bronchial asthma mice. Methods Mice spleen lymphocytes were divided into 5 groups: a health control group(group A),a health OVA group (group B),an asthma control group(group C),an asthma OVA group(group D),a treated group(group E).Based on the sequence in translation initiation region of the STAT6 mRNA 271～290 of mice,a segment of antisense oligonucleotid modifyed by sulfide was designed and synthetized.Astham model was duplicated with ovalbumin(OVA) absorbed to aluminum hydroxide. Spleen lymphocytes of mice were separated by the LymphoprepTM, then the STAT6 mRNA antisense vector was introduced into lymphocytes of mice with cationic liposome transfection methods. Observe the effect of antisense oligonucleotid on the expression of STAT6 mRNA and STAT6 protein in the spleen lymphocytes and IL-4 produced by spleen lymphocytes. The expression of STAT6 mRNA and STAT6 protein in the lymphocytes were determined by RT-PCR and immunocytochemistry respectively;IL-4 levels in the supernatants of spleen lymphocytes culture were determined by ELISA. Results The expression of STAT6 mRNA in the spleen lymphocytes was increased significantly in group D(2.26±0.52)than in group A (1.16±0.48) , group B (1.34±0.36) ,group C(1.20 ±0.24),group E(1.17 ±0.43)respectively,the difference being significant(P<0.01, respectively). However, after transfecting antisense oligonucleotid, the expression of STAT6mRNA in group E were significantly lower than those in group D (P<0.01). The expression of STAT6 protein in the spleen lymphocytes of treated group E decreased signifancantly.IL-4 increased markedly in the supernatants of lymphocytes culture in group D(43.68±5.5 pg/ml) than in group A(24.1±7.7 pg/ml) , group B(24.1±7.7 pg/ml), group C(24.1±7.7 pg/ml) ,group E(26.47±8.4 pg/ml) respectively ,the difference being significant(P<0.01, respectively).After transfecting antisense oligonucleotid,IL-4 levels in the supernatants of lymphocytes culture of group D was significantly different compared with group E(P<0.01). Conclusions The expression of STAT6 mRNA and STAT6 protein could be specialy inhibited by the STAT6 ASODN, consequently the IL-4 levels in the supernatants of lymphocytes culture of bronchial asthma mice decreased signifancantly.It is supported that the production of IL-4 be inhibited through preventing the transcription of STAT6 mRNA.The study provides foundation for antisense oligonucleotid therapeutic asthma.