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目的 研究巨噬细胞炎性蛋白 (MIP 1α)、血小板第 4因子 (PF4)及二者联合对造血干细胞化疗药物损伤的保护效应及其分子机制。方法 将骨髓、脐血单个核细胞及白血病细胞株HL 6 0分别予MIP 1α、PF4、MIP 1α +PF4、PBS预处理 48h ,再经柔红霉素 (DNR)孵育 2 4h后 ,用锥虫蓝 (台盼蓝 )拒染法测细胞活性 ,用流式细胞仪测细胞周期及CD34 + CD38- 细胞含量 ,细胞集落培养、免疫化学法测细胞P16、P2 7蛋白。结果 经MIP 1α及PF4预处理的骨髓和脐血单个核细胞 ,细胞活性、CD34+ CD38-细胞、集落形成能力均比对照组显著增加 (P <0 .0 5 )。MIP 1α、PF4组正常脐血及骨髓细胞S +G2 期百分率低于对照组 (P <0 .0 5 )。MIP 1α可上调细胞周期调控蛋白P16表达。PF4对P16、P2 7表达无影响。MIP 1α的保护作用强于PF4,但两者无协同效应。HL 6 0细胞P16、P2 7蛋白表达、细胞周期、细胞活性均不受MIP 1α、PF4的影响。结论 造血负调控因子MIP 1α、PF4能可逆性、选择性地保护正常造血细胞免受化疗药损伤 ,MIP 1α通过上调造血祖细胞G1 S期调控基因p16表达 ,使细胞阻滞于G0 期 ,提高细胞对周期特异性化疗药的耐受性。
Objective To investigate the protective effect of macrophage inflammatory protein (MIP 1α), platelet factor 4 (PF 4), and their combination on the damage of hematopoietic stem cell chemotherapy drugs and its molecular mechanism. METHODS: Bone marrow, cord blood mononuclear cells and leukemia cell line HL 60 were pretreated with MIP 1α, PF4, MIP 1α + PF4 and PBS for 48 h, and then incubated with daunorubicin (DNR) for 24 h. The activity of the cells was measured by blue staining (Typan blue). The cell cycle and CD34+CD38-cell contents were measured by flow cytometry. The cell P16 and P27 proteins were detected by cell colony culture and immunochemical assay. Results Both MIP 1α and PF4 pretreated bone marrow and umbilical cord blood mononuclear cells had significantly increased cell viability, CD34+ CD38-cells, and colony formation ability compared with the control group (P < 0.05). The percentage of normal cord blood and bone marrow cells in the MIP 1α and PF4 groups was lower than that in the control group (P < 0.05). MIP 1α upregulates the expression of cell cycle regulatory protein P16. PF4 had no effect on the expression of P16 and P27. The protective effect of MIP 1α was stronger than that of PF4, but there was no synergistic effect between the two. The expression of P16 and P27 protein, cell cycle and cell activity in HL 60 cells were not affected by MIP 1α and PF4. Conclusion The negative hematopoietic regulators MIP 1α and PF4 can selectively and selectively protect normal hematopoietic cells against chemotherapeutic agents. MIP 1α up-regulates the expression of p16 gene in G1 S phase of hematopoietic progenitor cells, which results in cell arrest in G0 phase and increases. The tolerance of cells to cycle-specific chemotherapeutic agents.