1，4-苯醌对人骨髓干细胞增殖能力的影响

Effects of 1, 4-benzoquinone on the proliferation activity of human bone marrow stem cells

目的探讨1，4-苯醌（1，4-benzoquinone，1，4-BQ）对人骨髓造血干细胞（hBM—HSCs）和人间充质干细胞（hBM-MSCs）增殖能力的影响。方法人骨髓样本采自健康自愿者。hBM—HSCs培养采用半固体甲基纤维素培养法，用集落形成实验（CFU）评价不同浓度1，4-BQ（10、25、50、100μmol／L）对人骨髓红细胞集落（CFU—E）／爆发式红细胞集落（BFU—E）、粒细胞-巨噬细胞系集落（CFU-GM）、粒细胞-红细胞-巨噬细胞-巨核细胞集落（CFU-GEMM）形成的影响。hBM—MSCs分离、培养扩增之后，采用噻唑蓝（MTT）实验评价不同浓度1，4-BQ（1、5、10、25、50、100、200、500、1000μmol／L）对hBM—MSCs增殖能力影响。结果CFU实验结果显示，25、50、100μmol／L1，4-BQ染毒组的集落形成数明显低于对照组，差异有统计学意义（P〈0．05）。10μmol／L1，4-BQ染毒组集落形成数与对照组的差异无统计学意义（P〉0．05）。MTT的结果显示，当1，4-BQ染毒24h，1，4-BQ浓度在50—200μmol／L时，细胞存活率下降趋势明显，存在剂量一反应关系；当染毒浓度≥200μmol／L时，细胞存活率小于5％，与对照组的差异有统计学意义（P〈0．05）；当染毒浓度≤25μmol／L时，细胞存活率与对照组的差异无统计学意义（P〉0．05）。结论1，4-BQ能影响hBM—HSCs的集落形成数和hBM．MSCs的相对存活率，1，4．BQ的血液毒性可能与其对hBM—HSCs和hBM—MSCs增殖能力的抑制有关。

Objective To explore the influence of 1,4-benzoquinone （1,4-BQ） on proliferation of human bone marrow haematopoietic stem cells （hBM-HSCs） and human bone marrow mesenehymal stem cells （hBM-MSCs）. Methods The bone marrow samples were collected from a healthy donor. Methyleellulose semi- solid culture medium was used to culture the mononuelear cells of bone marrow in different culture systems. Colony-forming unit （CFU） assay was utilized to evaluate the proliferation of hBM-HSCs exposed to 1,4-BQ at the doses of 10, 25, 50 and 100 μmol/L and to observe the influence of 1, 4-BQ on the Colony-forming unit- erythroid （CFU-E）/Burst-forming unit-erythroid （BFU-E）, Colony-forming unit-granulocyte, maerophage （CFU- GM）, Colony-forming unit-granulocyte, erythroid, maerophage, megakaryoeyte （CFU-GEMM） in hBM-MSCs. MTT assay was used to detect the proliferation of hBM-MSCs exposed to 1,4-BQ at the doses of 1, 5, 10, 25, 50, 100, 200, 500 and 1000 μmol/L for 24 h, respectively, after hBM-MSCs were isolated, cultured and expanded. Results The results of CFU assay indicated that numbers of CFU-E/BFU-E, CFU-GM and CFU-GEMM in 25, 50 and 100 μmol/L groups significantly decreased, as compared with control group （P〈O.05）. However, no significant difference was found between the 10 μmol/L group and the control group. The results of MTF assay showed that the cellular viability of hBM-MSCs exposed to 1, 4-BQ at the doses of 50-200 μmol/L for 24 h significantly decreased in a dose-depended manner. When the exposure dose was higher than 200μmol/L, the cellular viability of hBM-MSCs was lower than 5% which was significantly lower than that of control group （P〈0.05）. When the exposure dose was lower than 25 μmol/L, there was no significant difference of cellular viability between exposure group and control group （P〉0.05）. Conclusion The results of the present study demonstrated that 1, 4-BQ could inhibit the colony forming of hBM-HSCs and the relative viability of hBM-MSCs in vitro. The hematotoxieity induced by 1,4-BQ may be related to inhibiting the proliferation capacity of hBM-HSCs.