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目的 探讨核因子 κB(NF κB)在蛋白激酶C(PKC)对支气管哮喘T淋巴细胞增殖和凋亡调控的信号转导中的作用。方法 哮喘组和正常对照组的豚鼠以及急性发作期哮喘患者和正常对照者的外周血中分离出T淋巴细胞并分别加入PKC激动剂 12 肉豆蔻酰 13 乙酸佛波酯 (PMA)和NF κB抑制剂二硫代氨基甲酸吡咯烷 (PDTC)培养。用免疫组化法检测NF κB的表达 ,用四甲基偶氮唑盐微量比色法测定增殖反应 ,用原位末端终止法观测凋亡。结果 加入PMA培养的哮喘T淋巴细胞NF κB活化细胞百分比和增殖率与空白对照及加入PMA培养的正常T淋巴细胞相比均显著升高 (P <0 .0 1) ,而加入PDTC后以上指标均显著降低 (P <0 .0 1)。加入PMA培养的哮喘T淋巴细胞的凋亡指数与空白对照及加入PMA培养的正常T淋巴细胞相比均显著降低 (P <0 .0 1) ,加入PDTC后以上指标均显著升高 (P <0 .0 1)。T淋巴细胞NF κB活化细胞的百分比与增殖率均呈显著正相关 (r =0 .5 1～ 0 .72 ,P <0 .0 0 1) ,与凋亡指数均呈显著负相关 (r= 0 .5 5～ 0 .71,P <0 .0 0 1)。结论 T淋巴细胞PKC活化后导致增殖增加及凋亡减少的生物信号可能是通过激活NF κB来转导的。T淋巴细胞PKC NF κB信号转导途径的激活可能是哮喘的发病机制之一
Objective To investigate the role of nuclear factor κB (NFκB) in the signal transduction of protein kinase C (PKC) in the regulation of proliferation and apoptosis of T lymphocytes in bronchial asthma. Methods T lymphocytes were isolated from the peripheral blood of guinea pigs in the asthma group and the normal control group, as well as in acute exacerbation asthma patients and normal controls and were respectively treated with PKC agonist 12 myristoyl acetate (PMA) and NF κB Dithiocarbamic acid pyrrolidine (PDTC) culture. The expression of NF-κB was detected by immunohistochemistry. The proliferation reaction was measured by MTT assay. The apoptosis was observed by in situ terminal termination assay. Results The percentage of NF-κB activated cells and proliferation rate of asthmatic T lymphocytes cultured in PMA were significantly higher than those in blank control and PMA-supplemented normal T lymphocytes (P <0.01) Were significantly lower (P <0.01). The apoptotic index of asthmatic T lymphocytes cultured with PMA was significantly lower than that of blank control and normal T lymphocytes cultured in PMA (P <0.01), and all the above indexes were significantly increased after adding PDTC (P < 0 .0 1). The percentage of activated NF-κB in T lymphocytes was positively correlated with the proliferation rate (r = 0.51-0.72, P <0.01), and had a significant negative correlation with apoptotic index (r = 0 .5 5 ~ 0 .71, P <0. 0 0 1). Conclusions Biological signals that lead to increased proliferation and decreased apoptosis after PKC activation of T lymphocytes may be transduced by activating NF κB. The activation of PKC NF-κB signal transduction pathway in T lymphocytes may be one of the pathogenesis of asthma