红景天苷对阿糖胞苷诱导的骨髓间充质干细胞凋亡的影响

本研究旨在探讨红景天苷（salidroside）对阿糖胞苷（Ara-C）诱导的人骨髓间充质干细胞（humanbonemes-enchymalstemcells，hBMMSCs）凋亡影响及其机制。体外分离培养hBMMSC，流式细胞术鉴定免疫表型，特异性染色鉴定hBMMSC体外向成骨、成脂诱导分化能力，MTT法检测细胞增殖情况。实验分4组：对照组、Ara-C组、sali-droside组、Ara-C＋salidroside组；流式细胞术检测细胞凋亡变化；RT-PCR法检测BCL-2、BAXmRNA表达。结果表明，体外成功分离培养hBMMSC；细胞表达CIM4、CD29和HLA-ABC，不表达CD34、CD45和HLA．DR；成骨、成脂诱导21d，茜素红染色可见钙化结节，油红0染色有质滴出现；Ara-C对红景天苷处理的hBMMSC的增殖抑制程度较未经红景天苷处理的hBMMSC明显减低（P〈0．05）；在凋亡方面，Ara-C作用48h后hBMMSC的凋亡率显著高于对照组（P〈0．05），BCL-2基因表达下调，BAX表达上调；而中剂量红景天苷处理的细胞凋亡率较Ara-C组降低，BCL-2基因表达上调，BAX表达下调（P〈0．05）。结论：红景天苷可抑制Ara-C诱导的hBMMSC凋亡，其机制可能与BCL-2／BAX表达调控有关。

Effect of Salidroside on Apoptosis of Bone Marrow Mesenchymal Stem Cells Induced by Ara-C

The purpose of this study was to investigate the effect of salidroside on human bone marrow mesenchymal stem cell （hBMMSC） apoptosis induced by cytarabine C （Ara-C） and its mechanism, hBMMSC were cultured in vitro and isolated by Fircoll density gradient centrifugation ; cell surface antigens were measured by flow cytometry ; the osteo- genic and adipogenic differentiation of MSC was tested and evaluated by specific staining methods. The proliferation and apoptosis of cells exposed to Ara- C were detected by MTT and flow cytometry respectively. The expreiments were di- vided into 4 groups ： control group, Ara-C group, salidroside group and Ara-C ＋ salidroside group. The mRNA expres- sion of BCL-2 and BAX was assayed by RT-PCR. The results showed that the adherent cells displayed spindle and fibro- blast cell-like shape; the hBMMSC expressed CD44 ,CD71 and HLA-ABC, not expressed CD34 ,CD45 and HLA-DR; the hBMMSC successfully differentiated into osteogenic and adipogenic lineages, which showed mineralization with von Kossa staining. Furthermore, liquid vacuoles were detected by oil red O staining; Ara- C exhibited a less inhibitory effect on the proliferation of hBMMSC treated with slidroside. The apoptosis of hBMMSC treated with slidroside were significantly higher as compared with control group（ P 〈 0.05 ） ; RT-PCR results demonstrated that the BCL-2 expression Was significantly down regulated but BAX mRNA expressions was up-regulated in Ara- C group as compared with those in the control group. Salidroside significantly inhibited the apoptosis of MSC and reversed the mRNA expression of BCL- 2 and BAX. It is concluded that salidrosidd can inhibit the apoptosis of hBMMSC induced by Ara-C, its mechanism may be related with the regulation of BCL-2/BAX expression.