摘要
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.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
2012年第5期343-347,共5页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
浙江省科技厅科研院所专项资金(2008F1027)
浙江省自然科学基金(Y207394)
浙江省卫生厅优秀青年基金(2008QN002)
浙江省卫生厅支撑学科建设基金(11-ZC02)
关键词
1
4-苯醌
造血干细胞
间充质干细胞
集落形成单位
MTT
1,4-benzoquinone
Mesenehymal stem cells
Haematopoietie stem cells
Colony-forming units assay
MTT assay