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目的:探讨地龙提取物对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)增殖的影响。方法:全骨髓贴壁法体外分离培养大鼠BMSCs,用流式细胞术鉴定P3代BMSCs。将地龙配制成终浓度为500、50、5、0.5 mg/m L的提取物溶液作为药物干预组,以0 mg/m L组为空白对照组,CCK-8试剂盒检测细胞增殖情况。通过比较OD值判断地龙提取物对大鼠骨髓间充质干细胞增殖的影响,比较增殖率。结果:全骨髓贴壁法培养的P3代BMSCs表现为集落生长,经流式细胞术检测,CD90呈阳性表达,CD45呈阴性表达;CCK-8法检测结果为,剂量为0.5-50 mg/m L的药物干预组OD值较对照组显著升高;地龙浓度为5 mg/m L组OD值出现最大值,继续提高浓度OD值逐渐减小,当提取物浓度达到500 mg/m L时表现为细胞生长受到抑制。结论:培养的骨髓基质细胞在形态和表型上具备BMSCs的特性。地龙提取物在一定浓度范围内能促进BMSCs增殖。浓度为5mg/m L时促进BMSCs增殖作用最为明显,浓度为500 mg/m L时会抑制BMSCs的增殖。
Objective: To investigate the effect of earthworm extract on the proliferation of bone marrow mesenchymal stem cells (BMSCs). Methods: BMSCs were isolated and cultured in vitro by whole bone marrow adherent method. P3-generation BMSCs were identified by flow cytometry. The cucumbers were prepared into 500, 50, 5, 0.5 mg / mL final extract solution as drug intervention group, 0 mg / mL group as blank control group, CCK-8 kit to detect cell proliferation. By comparing the OD value to determine the effect of earthworm extract on the proliferation of rat bone marrow mesenchymal stem cells, compare the proliferation rate. Results: The BMSCs of P3 generation cultured by whole bone marrow adherent method showed colony growth. CD90 expression was positive by flow cytometry and negative by CD45. The CCK-8 assay showed that the dose of BMSCs was 0.5-50 mg / m L in the drug intervention group OD value was significantly higher than that of the control group; the dexamethasone concentration of 5 mg / m L group OD value maximum, continue to increase the concentration of OD value decreases gradually, when the extract concentration reaches 500 mg / m L The performance of cell growth was inhibited. Conclusion: The cultured bone marrow stromal cells possess the characteristics of BMSCs in morphology and phenotype. Earthworm extract can promote the proliferation of BMSCs in a certain concentration range. The proliferation of BMSCs was most obvious when the concentration was 5mg / m L, and inhibited the proliferation of BMSCs when the concentration was 500 mg / m L.