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目的:进一步探讨获得性再生障碍性贫血(AA)发病机制及健脾补肾活血方生血合剂治疗AA的疗效机制。方法:以环孢素为对照,免疫介导AA小鼠为研究对象,采用定量PCR(Q-PCR)、免疫组织化学等方法分析生血合剂干预前后AA小鼠骨髓核因子κB(NF-κB)mRNA水平、蛋白表达及活性变化。结果:AA小鼠骨髓单个核细胞NF-κB mRNA水平未见明显改变,与正常对照比无显著差异(P>0.05);骨髓细胞NF-κB/p65蛋白表达较正常组增加,p-IκBα表达明显下降,生血合剂干预后NF-κB/p65蛋白表达减少,而p-IκBα表达明显提高;与之相比,环孢素作用后NF-κB/p65蛋白及p-IκBα均急剧下降。结论:NF-κB蛋白表达异常及活性改变可能参与了免疫介导AA小鼠模型的形成;生血合剂可能通过调节NF-κB的表达及活性发挥改善免疫介导AA小鼠造血功能的作用。
Objective: To further investigate the pathogenesis of acquired aplastic anemia (AA) and the therapeutic mechanism of Jianpi Bushen Huoxue Fangsheng Xue mixture in the treatment of AA. METHODS: Using CsA as control, immune-mediated AA mice were studied. Q-PCR and immunohistochemistry were used to analyze the bone marrow nuclear factor-κB (NF-κB) in AA mice before and after intervention with Shengxue mixture. mRNA levels, protein expression, and activity changes. Results: There was no significant change in NF-κB mRNA level in bone marrow mononuclear cells of AA mice. There was no significant difference compared with normal control (P>0.05). The expression of NF-κB/p65 protein in bone marrow cells was increased compared with the normal group, and the expression of p-IκBα was detected. After a significant decline, the expression of NF-κB/p65 protein was decreased and the expression of p-IκBα was significantly increased after intervention with Shengxue Mixture. In contrast, the NF-κB/p65 protein and p-IκBα all decreased rapidly after cyclosporine treatment. Conclusion: The abnormal expression of NF-κB protein and its activity may be involved in the formation of immune-mediated AA mouse model; Shengxue Mixture may improve the immune-mediated hematopoietic function of AA mice by regulating the expression and activity of NF-κB.