PKC信号传导对ITP患儿T细胞核因子-κB活化的调控研究

Signal transduction of protein kinase C regulating the activation of nuclear factor-кB in T lymphocytes from children with idiopathic thrombocytopenic purpura

目的探讨蛋白激酶C(PKC)对ITP患儿T细胞核因子-кB(NF-кB)活化的调控作用。方法分别从35例特发性血小板减少性紫癜(ITP)患儿及30例健康儿童的外周血中分离出T细胞并分成3组培养,即空白对照组,加入PKC的激活剂PMA的PMA组,同时加入PKC激活剂PMA与抑制剂H-7的PMA+H-7组,分别用免疫组化法和Westernblot检测NF-кB和抑制蛋白кB(I-кB)。结果ITP患儿的PMA组NF-кB活化的细胞百分比显著高于空白对照组和正常儿童的PMA组(P<0.05),且I-кB水平显著低于上述两组(P<0.05),而ITP的PMA+H-7组NF-кB活化的细胞百分比显著低于ITP的PMA组,且I-кB水平显著高于该组(P<0.05)。结论ITP患儿T细胞NF-кB的活化可能受PKC信号传导的调控,T淋巴细胞PKC-NF-кB信号传导途径的激活可能是ITP的发病机制之一。

Objectives To explore the role of protein kinase C （PKC） in regulating the activation of nuclear factor-κB （NF-κB） in T lymphocyte from idiopathic thromboeytopenic purpura （ITP） children. Methods Peripheral blood from ITP children （n = 35） and healthy children （n = 30） were collected. T lymphocytes were purified and divided into three groups separately： control group,phorbol 12-myristate 13-acetate （PMA） treated group,and PMA ＋ H-7 treated group. The expression of NF-κB and inhibitor protein-κB （I-κB） was detected by immunohistochemical staining and Western blot,respectively. Results The percentage of cells with active NF-κB was significantly higher and the expression level of I-κB was significantly lower in ITP-PMA group than that in ITP-control group and healthy-PMA group respectively （all P 〈 0.05）. The percentage of cells with active NF-κB was significantly lower and the expression level of I-κB was significantly higher in ITP -PMA ＋ H-7 group than that in ITP-PMA group（both P 〈 0,05）. Conclusions It revealed that the activation of NF-κB in ITP T lymphocyte may be regulated by PKC, and the activation of PKC- NF-κB signal pathway of T Lymphocyte may play an important role in the pathogenesis of ITP.