摘要
探讨了刀豆蛋白A(ConA)活化巨噬细胞 (M)的机理。以ConA 5 0 0 μg腹腔注入预处理小鼠 4 8h ,并以PBS作对照 ,免疫组织化学染色法检测诱导型一氧化氮合成酶 (iNOS)的表达 ;分别以原位杂交及流式细胞仪检测NF κBP6 5mRNA与NF κBRelA蛋白的表达 ;RT PCR了解TNF α、IL 1βmRNA的表达情况。ConA刺激后 ,iNOS在胞浆中表达增多 ;ConA处理组NF κBP6 5mRNA及NF κBRelA蛋白的表达均增加 ,RT PCR方法测到TNF α、IL 1βmRNA的表达水平也显著提高 (P <0 0 1)。ConA作用于M时 ,可能通过活化NF κB使iNOS、TNF α及IL 1β表达增强 。
To explore signal transduction during the process of macrophages (M) activation stimulated by ConA. Peritoneal macrophages were collected from mice that had received ConA or phosphate-buffered saline( PBS). The cells were allowed to adhere for 2 h at 37℃ and the monolayers of Mφ were cultured for further 24h then fixed for the immunohistochemical stain assay of iNOS. Nuclear factor kappa B (NF-κB) P65 mRNA and NF-κB RelA protein measured respectively by in situ hybridization and FACS. Total RNA was isolated and RT-PCR amplification was carried out using specific primers for TNF-α and IL-1β. iNOS were not or just poorly expressed in normal peritoneal macrophages, but we found iNOS strong positive in those treated with ConA in vivo. We also get similar results from transcriptional level of NF-κB P65 mRNA and NF-κB RelA protein expression. mRNA expression of TNF-α and IL-1β in macrophages treated with ConA was higher than that of normal ones. Macrophages treated with ConA in vivo showed that NF-κB activation was mediated by iNOS mRNA expression and proinflammatory cytokines production.
出处
《基础医学与临床》
CSCD
北大核心
2003年第6期661-665,共5页
Basic and Clinical Medicine
