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作 者:李国平[1] 刘志刚[1] 冉丕鑫[2] 丘劲[1] 钟南山[2]
机构地区:[1]深圳大学生命科学学院微生物基因工程重点实验室,广东深圳518060 [2]广州医学院广州呼吸病研究所,广东广州510182
出 处:《细胞与分子免疫学杂志》2004年第5期611-614,共4页Chinese Journal of Cellular and Molecular Immunology
基 金:国家高技术研究发展计划 (863)资助项目 (No .2 0 0 2AA2 1 4 0 1 1 );国家自然科学基金资助项目 (No .30 2 71 2 2 6 ;No.39860 0 71 )
摘 要:目的 :研究卵白蛋白 (OVA)在哮喘小鼠脾细胞增殖中信号转导蛋白和转录激活因子 5 (STAT5 )的变化 ,探讨STAT5在OVA诱导哮喘小鼠脾细胞增殖中的作用。方法 :经腹腔注射与雾化吸入OVA诱发小鼠哮喘发作。激发后分离小鼠脾细胞 ,采用MTT比色法测定经OVA诱导的脾细胞增殖 ;用双染免疫组化法检查脾细胞中STAT5磷酸化与STAT5抗原的定位。用电泳迁移率变动分析 (EMSA)测定STAT5与DNA探针的结合力。结果 :OVA在体外能明显诱导哮喘小鼠脾细胞增殖 (平均A值为 0 .5 4 3± 0 .112 ) ,呈剂量依赖性 ,与对照组相 (平均A值为 0 .2 0 3± 0 .0 32 )比较差异明显 (P <0 .0 1)。OVA诱导 1、3h ,能诱导哮喘小鼠脾细胞中的STAT5磷酸化 ,脾细胞中出现STAT5 DNA结合带。结论 :OVA能诱导哮喘小鼠脾细胞增殖和脾细胞中的STAT5磷酸化 ,增加STAT5与DNA探针的结合力 。AIM: To investigate the changes of signal transducer and transcriptional activator 5 (STAT5) in proliferative splenocyte of asthma mice. METHODS: BALB/c mice were induced to develop asthma by intraperitoneal injection and then nebulized aspiration of OVA. The splenocytes were isolated, and the proliferation of the splenocytes was detected by MTT colorimetry . The phosphorylation of STAT5 and its location were examined by double immunohistochemical staining and the binding of STAT5 with DNA was determined by electrophoretic mobility shift assay (EMSA). RESULTS: OVA could pronouncedly induce the splenocyte proliferation of asthma mice in dose-dependent manner. Stimulation with OVA for 1 and 3 hours could lead to phosphorylation of STAT5 in splenocytes from asthma mice and increase the binding of STAT5 with DNA, suggesting that STAT5 signal pathway plays an important role in splenocyte proliferation of asthma mice induced with OVA.
关 键 词:哮喘 淋巴细胞增殖 信号转导蛋白和转录激活因子5 卵白蛋白
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