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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 ,说明STAT5信号途径在OVA诱导的哮喘小鼠脾细胞增殖中起重要作用
Objective: To investigate the changes of signal transducer and activator of transcription 5 (STAT5) in ovalbumin (OVA) - induced splenocyte proliferation in asthmatic mice and the role of STAT5 in the proliferation of splenocytes induced by OVA in asthmatic mice. Methods: Asthma attack was induced in mice by intraperitoneal injection and inhalation of OVA. Spleen cells were isolated after challenge, and the splenocyte proliferation induced by OVA was determined by MTT colorimetric assay. The phosphorylation of STAT5 and the localization of STAT5 antigen in spleen cells were examined by double staining immunohistochemistry. The binding of STAT5 to the DNA probe was determined by electrophoretic mobility shift assay (EMSA). Results: OVA could obviously induce the proliferation of splenocytes in asthmatic mice in vitro (average A value was 0.54 3 ± 0.112) in a dose-dependent manner, with the average phase A value of 0. 023 ± 0 .0 32) was significantly different (P <0. 01). OVA induced 1,3h, can induce STAT5 phosphorylation in spleen cells of asthmatic mice, and STAT5 DNA binding band in spleen cells. CONCLUSION: OVA can induce the proliferation of splenocytes in asthmatic mice and the phosphorylation of STAT5 in splenocytes, and increase the binding ability of STAT5 to DNA probes. This indicates that STAT5 signaling plays an important role in the proliferation of OVA-induced asthmatic mice