人骨髓间充质干细胞上清液对体外肝星状细胞增殖周期及MMP-1表达的影响

目的 研究骨髓间充质干细胞(BMSC)对肝星状细胞(HSC)增殖周期及基质金属蛋白酶-1(MMP-1)活性的影响,以探讨其防治肝纤维化的作用机制.方法 采用密度梯度离心法分离鉴定人BMSC,收集原代培养7d的BMSC培养上清,加入HSC中培养24 h及48 h.通过流式细胞仪观察HSC增殖周期,ELISA方法检测MMP-1浓度.结果 与HSC单独培养组相比,共培养48 h组G1期细胞显著增加(P＜0.01),S期细胞显著减少(P＜0.01),并且共培养组MMP-1表达明显增加,与对照组相比差异均具有统计学意义(P＜0.05).结论 BMSC可抑制HSC的增殖,并且可能通过增加MMP-1的产生,从而起到抗纤维化的作用.

Effect of human bone marrow mesenchymal stem cell supernatant on proliferation cycle and MMP-1 expression of hepatic stellate cells

Objective To study the effect of secreted factors from bone marrow mesenchymal stem cells(BMSCs) on hepatic stellate cell(HSC) proliferation and expression of matrix metalloproteinase-1(MMP-1),and to explore the underlying mechanisms of BMSC-mediated prevention and treatment of liver fibrosis.Methods Density gradient centrifugation was used to isolate BMSCs.After seven days of growth in culture,the BMSC culture supernatant was collected and added to HSCs.After 24 h or 48 h of culture,flow cytometry was carried out to determine the HSC cell cycle stage and enzyme-linked immunosorbent assay was performed to detect the concentration of MMP-1.HSCs cultured without BMSC supernatant served as controls.Results At 48 h,HSCs cultured with BMSC supernatent showed a significant increase in G1 phase cells(P<0.01) and significant reduction in S phase cells(vs.control HSCs;P<0.01).Additionally,expression of MMP-1 was significantly higher in HSCs cultured with BMSC supernatant(vs.control HSCs;P<0.05).Conclusion BMSC secreted factors may play a role in anti-fibrosis by inhibiting HSC proliferation and increasing production of MMP-1.