Xiong W, Xu Y, Zhang Z
Department of Respiratory Medicine, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2001 Jun;24(6):355-9.
To explore the role of nuclear factor-kappaB (NF-kappaB) in the signal pathway of protein kinase C (PKC) regulating the expression of Th2 cytokines, interleukin 4 (IL-4) and IL-5, by T lymphocyte in asthma.
16 guinea pigs were randomly divided into 2 groups, normal control and asthmatic group. 16 asthmatic patients and 16 normal control persons also participated in the study. T lymphocytes were isolated and purified from blood of each guinea pig and each person and divided into 3 groups: cells of 1st group were control group, cells of 2nd group were stimulated with phorbol 12-myristate 13-acetate (PMA), cells of 3rd group were stimulated with PMA and pyrrolidine dithiocarbamate (PDTC). The expression of NF-kappaB were observed by immunocytochemical staining, the expression of IL-4 and IL-5 mRNA were observed by in situ hybridization staining, IL-4 and IL-5 protein in supernatants were measured by ELISA.
The percentage of cells of active NF-kappaB, the percentage of positive cells of IL-4 and IL-5 mRNA, IL-4 and IL-5 protein in supernatants of asthmatic T lymphocyte stimulated with PMA were significantly higher than those of asthmatic T lymphocyte stimulated without PMA (q = 8.44 approximately 38.66, P < 0.01) and those of normal control T lymphocyte stimulated with PMA (q = 8.11 approximately 40.12, P < 0.01), and were significant reduced by PDTC (q = 6.50 approximately 35.63, P < 0.01). There were good positive correlation between the percentage of cells of active NF-kappaB and the percentage of positive cells of IL-4 and IL-5 mRNA (r = 0.60 approximately 0.82, P < 0.001, respectively), and also good positive correlation between the percentage of active NF-kappaB and IL-4 and IL-5 protein in supernatants (r = 0.42 approximately 0.70, P < 0.005 or < 0.001, respectively).
The active PKC of asthmatic T lymphocyte increasing the expression of IL-4 and IL-5 may be mediated by activating NF-kappaB, the activation of PKC-NF-kappaB signal pathway of T lymphocyte NF-kappab may play an important role in the pathogenesis of asthma.
探讨核因子-κB（NF-κB）在蛋白激酶C（PKC）调节哮喘患者T淋巴细胞中Th2细胞因子白细胞介素4（IL-4）和白细胞介素5（IL-5）表达的信号通路中的作用。
将16只豚鼠随机分为正常对照组和哮喘组。16例哮喘患者和16例正常对照者也参与了研究。从每只豚鼠和每个人的血液中分离并纯化T淋巴细胞，分为3组：第1组细胞为对照组，第2组细胞用佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）刺激，第3组细胞用PMA和吡咯烷二硫代氨基甲酸盐（PDTC）刺激。通过免疫细胞化学染色观察NF-κB的表达，通过原位杂交染色观察IL-4和IL-5 mRNA的表达，用ELISA法检测上清液中IL-4和IL-5蛋白。
用PMA刺激的哮喘T淋巴细胞上清液中活性NF-κB细胞百分比、IL-4和IL-5 mRNA阳性细胞百分比、IL-4和IL-5蛋白水平均显著高于未用PMA刺激的哮喘T淋巴细胞（q = 8.44～38.66，P < 0.01）及用PMA刺激的正常对照T淋巴细胞（q = 8.11～40.12，P < 0.01），且PDTC可使其显著降低（q = 6.50～35.63，P < 0.01）。活性NF-κB细胞百分比与IL-4和IL-5 mRNA阳性细胞百分比之间呈良好的正相关（r分别为0.60～0.82，P < 0.001），活性NF-κB细胞百分比与上清液中IL-4和IL-5蛋白之间也呈良好的正相关（r分别为0.42～0.70，P < 0.005或< 0.001）。
哮喘T淋巴细胞中活化的PKC增加IL-4和IL-5的表达可能是通过激活NF-κB介导的，T淋巴细胞NF-κB的PKC-NF-κB信号通路激活可能在哮喘发病机制中起重要作用。
