基于PP2A/AKT通路探讨扶正祛毒汤对急性髓系白血病-完全缓解患者CD34^(+)细胞源DC激发T细胞的影响

Exploration on the effects of Fuzheng Qudu Decoction interventing AML-CR patients' CD34~+ cells source DC activate T cells based on PP2A/AKT signaling pathway

目的:探讨扶正祛毒汤治疗急性髓系白血病-完全缓解(AML-CR)的可能作用机制。方法:运用血清药理学、Ficoll密度梯度离心、免疫磁珠阳性选择、细胞培养、流式细胞分析、Western Blot和RT-qPCR等技术检测DC表面CD1a、CD80、CD83、CD86、HLA-DR的表达,DC激发T细胞中CTLA-4、PP2A、AKT、p-AKT蛋白和mRNA的表达。结果:扶正祛毒汤含药血清能诱导AML-CR患者CD34^(+)细胞形成典型的DC形态,显著增加DC表面CD1a、CD80、CD83、CD86、HLA-DR的表达(P<0.05),上调DC激发T细胞中PP2A蛋白及mRNA的表达(P<0.05),下调CTLA-4、AKT蛋白及mRNA的表达(P<0.05),下调p-AKT蛋白表达(P<0.05)。结论:扶正祛毒汤治疗AML-CR的作用机制可能与该方能够促使AML-CR患者CD34^(+)细胞源DC成熟,抑制CTLA-4与CD80、CD86的结合,有效激活T细胞,调控PP2A/AKT信号通路,进而杀伤白血病细胞有关。

Objective:To explore the possible mechanism of Fuzheng Qudu Decoction in the treatment of acute myeloid leukemi-complete remission(AML-CR).Methods:Serum pharmacology,Ficoll density gradient centrifugation,positive selection with immunomagnetic beads,cell culture,flow cytometric analysis,Western Blot and RT-qPCR were used to determine the expression of CD1a,CD80,CD83,CD86 and HLA-DR on the surface of DC;and determine the expression of CTLA-4,PP2A,AKT,p-AKT protein and mRNA in DC activating T cells.Results:Serum containing drugs from Fuzheng Qudu Decoction induced the formation of typical DC morphology in CD34~+ cells from AML-CR patients,significantly increased the expression of CD1a,CD80,CD83,CD86 and HLA-DR on the surface of DC(P<0.05);upregulated the expression of PP2A protein and mRNA(P<0.05),downregulated the expression of CTLA-4,AKT protein and mRNA(P<0.05),and downregulated the expression of p-AKT protein in DC activating T cells(P<0.05).Conclusion:The mechanism of Fuzheng Qudu Decoction in treating AML-CR may be related to its ability to promote the maturation of CD34~+ cell-derived DC in AML-CR patients,inhibit the combination of CTLA-4 with CD80 and CD86,effectively activate T cells,regulate PP2A/AKT signaling pathway,and then kill leukemia cells.