目的 观察阻断Notch1信号通路对慢性乙肝患者外周血单个核细胞(PBMC)Foxp3 mRNA表达以及细胞毒性T淋巴细胞(CTL)细胞毒活性的影响,探讨Notch1信号通路在乙型肝炎慢性化过程中可能的作用机制.方法 抗Notch1抗体阻断Notch1信号通路,利用实时荧光定量PCR(real-time PCR)检测其外周单个核细胞Foxp3 mRNA的表达情况,并用乳酸脱氢酶释放法检测CTL对靶细胞HepG2.2.15细胞毒活性的影响.结果 用抗体阻断后,慢性乙型肝炎患者外周单个核细胞Foxp3 mRNA表达显著减弱,且CTL细胞毒活性明显增强,与阴性对照组和空白对照组相比差异有统计学意义(P均<0.01).结论 阻断Notch1信号通路可减少慢性乙型肝炎患者外周单个核细胞Foxp3 mRNA的表达,增强CTL细胞毒活性,说明Notch1信号通路可能是HBV持续感染的一个因素,且可能与Foxp3有关.%Objective To investigate the influence of blockage of Notch1 signaling on expression of Foxp3 mRNA and cytotoxic T-lymphocyte (CTL) cytotoxicity in peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis B, and analyze the possible mechanism of Notch1 signaling pathway during chronic HBV infection. Methods In peripheral blood of patients with chronic hepatitis B, after blocking Notch1 signaling with anti-Notch1, the expression of Foxp3 mRNA was examined by real-time PCR, and CTL cytotoxicity was measured by Cytotoxicity Detection Assay (LDH). Results After blocking Notch1 signaling with anti-Notch1 , the expression of Foxp3 mRNA in peripheral blood of patients with chronic hepatitis B was decreased evidently, which was lower than that in the negative control group or the blank control group (P ＜ 0. 01 ). CTL cytotoxicity was enhanced remarkably, which was higher than that in the both control groups (P ＜ 0. 01 ). Conclusion Blocking Notch1 signaling could eliminate the expression of Foxp3 mRNA and enhance CTL cytotoxicity in peripheral blood of patients with chronic hepatitis B. It suggests Notch1 signaling pathway may be one of causes of chronic hepatitis B virus infection, and perhaps related with Foxp3.
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