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目的观察肝再生增强因子(ALR)对外周血单核细胞增殖时ERK的影响,以探明ALR免疫抑制相关机理。方法梯度离心法分离健康人外周单核细胞,用ConA 5μg/ml刺激细胞增殖,选定最佳研究时间;观察不同剂量ALR抑制功能,选用最佳抑制剂量;利用Western blot检测ALR抑制细胞增殖时ERK的磷酸化改变。结果 ConA刺激细胞增殖最佳时间是60 h,ALR能抑制细胞增殖,并呈剂量依赖关系,30μg/ml ALR抑制效果最显著;ALR对单核细胞无直接增殖作用。ConA能引起ERK含量和磷酸化明显增加,ALR则抑制ConA对ERK的刺激,以抑制ERK2最明显。结论 ALR可能通ERK的含量和抑制ERK2的磷酸化抑制细胞增殖。
Objective To observe the effect of augmenter of liver regeneration (ALR) on ERK in the proliferation of peripheral blood mononuclear cells in order to find out the mechanism of ALR immunosuppression. Methods Peripheral blood mononuclear cells (PBMCs) were isolated by gradient centrifugation. Cell proliferation was stimulated with ConA 5μg / ml and the optimal time was selected. The inhibitory effect of different doses of ALR was observed and the optimal inhibitory concentration was determined. Phosphorylation of ERK changes. RESULTS: The optimal time for ConA to stimulate cell proliferation was 60 h. ALR inhibited cell proliferation in a dose-dependent manner. The inhibitory effect of ALR at 30 μg / ml was the most significant. ALR did not directly proliferate monocytes. ConA can cause a significant increase in ERK content and phosphorylation, while ALR inhibits the stimulation of ERK by ConA to suppress the most obvious ERK2. Conclusion ALR may inhibit the proliferation of ERK2 cells through inhibiting ERK2 phosphorylation.