Experimental Study on Treatment of Bronchial Asthma With Antisense Oligonucleotid / 复旦学报（医学版）

ABSTRACT

Purpose　To explore the possibility and the effect of therapeutic bronchial asthma by antisense oligonucleotid.　Methods　Based on the IL-5 cDNA sequence of mouse,a segment of antisense oligonucleotid was designed and synthetized.5′-labeling of antisense oligonucleotid was signed by T4 PNK in order that the efficiency of stearylamine liposome in transfe-ting antisense oligonucleotid can be evaluated. Astham model was duplicated with ovalbumin (OVA) absorbed to aluminum hydroxide. T lymphocytes of mice were separated by nylon fiber method,then T lymphocytes transfected a different content of antisense oligonucleotid with stearylamine phys. positive liposome were cultured respectively in order to observe the effect of antisense oligonucleotid on IL-5 produced by T lymphocytes. IL-5 levels in the supernatants of T lymphocytes culture were determined by ELISA.　Results 　Stearylamine liposome could markedly increase the efficiency of antisense oligonucleotid transfection. The efficiency of antisense oligonucleotid transfection was the best at 1∶15 m/m(antisense oligonucleotid and SA liposome) and it was increased approximately 12 times.In healthy and asthma Balb/c mice, IL-5 was not detected in the supernatants of T lymphocytes culture without challenge with OVA.However,IL-5 was increased markedly in the supernatants of T lymphocytes culture challenged with OVA. After transfecting a different concentration antisense oligonucleotid, IL-5 levels in the supernatants of T lymphocytes culture were significantly lower than those in control cells without antisense oligonucleotid transfection. IL-5 levels decreased from (44.60±6.23) to (30.70±7.362),(17.20±6.181) and(8.16±2.34)pg/ml respectively. And IL-5 synthesis was inhibited by 31.17% , 61.43% and 81.7% respectively. 　Conclutions　IL-5 synthesis could be obviously inhibited by antisense oligonucleotid and showed a markedly relation between quantitative and effect. It is supported that the production of IL-5 be inhibited through preventing the transcription of IL-5 from T lymphocytes. The study provides foundation for antisense gene therapeutic asthma.