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目的:探讨黄芪多糖对多发性骨髓瘤(multiple myeloma,MM)患者骨髓间充质干细胞(bone mesenchymal stem cells,BMSC)增殖与细胞因子表达的影响。方法:采用密度梯度离心法分离MM患者骨髓间充质干细胞(MM-BMSC)及正常人骨髓间充质干细胞(ND-BMSC)。取第3代BMSC,测定细胞的生长曲线及细胞因子的表达。分别采用MTT法、细胞流式术检测不同浓度APS对MM-BMSC的细胞增殖、细胞周期的影响,ELISA法测定IL-1β与IL-6的表达水平。结果:成功分离得到MM-BMSC和ND-BM-SC,细胞生长曲线显示MM-BMSC增殖缓慢,倍增时间为86 h,而ND-BMSC的倍增时间为60 h,两者相比差异具有统计学差异(P<0.05)。基础正常情况下,MM-BMSC与ND-MSC分泌的IL-1β、IL-6的水平有明显差异(P<0.05)。APS可促进MM-BMSC增殖,其中1 mg/mL APS的作用最为明显(P<0.01)。MM-BMSC基本都处于静止期,1 mg/mL APS处理的MM-BMSC处于G0/G1期细胞比例较其明显降低(P<0.05),S,G2/M期细胞比例较其明显增高(P<0.05),且1 mg/mL APS可显著下调MM-BMSC分泌IL-1β及IL-6的水平(P<0.05)。结论:多发性骨髓瘤患者骨髓间充质干细胞IL-1β、IL-6分泌水平异常,1 mg/mL黄芪多糖可促进骨髓间充质干细胞的增殖并下调其IL-1β,IL-6的分泌。
Objective: To investigate the effect of astragalus polysaccharide on the proliferation and cytokine expression of bone marrow mesenchymal stem cells (BMSCs) in patients with multiple myeloma (MM). METHODS: Mesenchymal stem cells (MM-BMSCs) and normal human bone marrow mesenchymal stem cells (ND-BMSCs) were isolated by density gradient centrifugation. 3rd generation BMSCs were used to determine the cell growth curve and cytokine expression. The effects of different concentrations of APS on cell proliferation and cell cycle of MM-BMSC were detected by MTT assay and cell flow assay respectively. The expression levels of IL-1β and IL-6 were determined by ELISA. RESULTS: MM-BMSC and ND-BM-SC were successfully isolated. The cell growth curve showed that MM-BMSC proliferation was slow, the doubling time was 86 h, and the doubling time of ND-BMSC was 60 h. The difference was statistically significant. Difference (P<0.05). Under normal conditions, the levels of IL-1β and IL-6 secreted by MM-BMSC and ND-MSCs were significantly different (P<0.05). APS could promote the proliferation of MM-BMSCs, of which the effect of 1 mg/mL APS was the most obvious (P<0.01). MM-BMSCs were all in quiescent phase. The proportion of MM-BMSCs treated with 1 mg/mL APS in G0/G1 phase was significantly lower than that of MM-BMSCs (P<0.05), and the percentage of S,G2/M phase cells was significantly higher (P<0.05). <0.05), and 1 mg/mL APS could significantly down-regulate the levels of IL-1β and IL-6 secreted by MM-BMSCs (P<0.05). Conclusion: The levels of IL-1β and IL-6 in BMSCs were abnormal in patients with multiple myeloma. 1 mg/mL astragalus polysaccharide could promote the proliferation of bone marrow mesenchymal stem cells and down-regulate the secretion of IL-1β and IL-6. .