STAT6 ASODN对哮喘小鼠脾淋巴细胞影响的实验研究  

EXPERIMENATAL STUDY ON THE EFFECT OF STAT6 ANTISENSE OLIGONUCLEOTIDE ON SPLEEN LYMPHOCYTES OF MICE WITH BRONCHIAL ASTHMA

作　　者：赵鹰[1] 弓清梅[1] 田新瑞[1] 任文霞[1] 李建强[1] 刘卓拉[1] 

机构地区：[1]山西医科大学第二医院呼吸科,太原030001

出　　处：《中国组织化学与细胞化学杂志》2006年第2期195-200,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：山西省归国留学人员基金(200429)

摘　　要：目的研究STAT6反义寡核苷酸对哮喘小鼠脾淋巴细胞的影响作用。方法实验细胞分组:正常鼠空白组(A组)、正常鼠OVA组(B组)、哮喘空白组(C组)、哮喘OVA组(D组)、哮喘治疗组(E组)。正常设计并人工合成一段互补于小鼠STAT6 mRNA翻译起始区271-290的反义寡核苷酸片段,全链硫代修饰。用卵白蛋白和氢氧化铝复制哮喘模型,用淋巴细胞分离液分离脾淋巴细胞,进行体外培养并导入由阳离子脂质体转染剂Geneshuttle携带的反义寡核苷酸,观察反义寡核苷酸的转染对脾淋巴细胞STAT6蛋白表达水平及细胞培养上清中IL-4分泌水平的影响。免疫细胞化学观察脾淋巴细胞中STAT6蛋白的表达水平,同时采用酶联免疫吸附(ELISA)法测定脾细胞培养上清液中IL-4的浓度。结果D组细胞STAT6蛋白表达明显高于其余各组,均具有显著性差异(P均<0.01),STAT6 ASODN转染后,E组细胞该蛋白的表达量明显下降(P<0.01);D组脾淋巴细胞培养上清中IL-4分泌水平明显高于其余各组,均具有显著性差异(P均<0.01);STAT6 ASODN转染后,E组培养上清中IL-4分泌水平显著低于D组(P<0.01)。结论STAT6 ASODN可特异性抑制哮喘鼠脾淋巴细胞中STAT6蛋白的表达,并可特异性抑制脾淋巴细胞中IL-4的分泌,为反义基因技术治疗哮喘提供了依据。Objective To explore the effect phocytes of mice with bronchial asthma. Methods of STAT6 antisense oligonucleotide on the spleen lym- Mouse 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）, and a treated group （group E）. Based on the sequence in the translation initiation region of the STAT6 mRNA 271-290 of mice, a segment of antisense oligonucleotide was designed and synthesized. The asthma model was duplicated with ovalbumin （OVA） absorbed to aluminum hydroxide. Spleen lymphocytes of mice were separated, then the STAT6 mRNA antisense vector was introduced into the lymphocytes with cationic liposome transfection methods. The effect of antisense oligonucleotid on the expression of STAT6 protein in the spleen lymphocytes and IL-4 produced by spleen lymphocytes was observed. The expression of STAT6 protein in the lymphocytes was determined by immunocytochemistry, and IL-4 levels in the supernatants of spleen lymphocyte culture were determined by ELISA. Results The expression of STAT6 protein in the spleen lymphocytes was increased significantly in group D than in groups A, B, C and E （P〈0. 01, respectively）. However, after the transfection of antisense oligonucleotide, the expression of STAT6 protein in group E was significantly lower than that in group D （P〈0.01）. IL-4 increased markedly in the supernatants of lymphocytes culture in group D than in the other groups, the difference being significant （P〈0.01, respectively）. After the transfection of antisense oligonucleotide, IL-4 levels in the supernatants of lymphocyte culture of group E were significantly different from those of group D （P〈0.01）. Conclusion The expression of STAT6 protein were specifically inhibited by STAT6 ASODN, and simultaneously the IL-4 levels in the supernatants of the lymphocyte culture of bronchial asthma mice decreased significantly. The results suggest that both

关 键 词：反义寡核苷酸 支气管哮喘 白介素-4 信号转导和转录激活因子6 

分 类 号：R332.35[医药卫生—人体生理学]