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  

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

作　　者：魏玉萍 白海[2] 孙延庆[3] 包慎 茜瑞 刘琳[2] 

机构地区：[1]宁夏人民医院血液科,宁夏银川750012 [2]兰州军区兰州总医院血液科,甘肃省干细胞与基因药物重点实验室,甘肃兰州730050 [3]甘肃省人民医院血液科,甘肃兰州730000

出　　处：《中国实验血液学杂志》2013年第6期1572-1577,共6页Journal of Experimental Hematology

基　　金：军队高新技术重大项目(编号2010GXJS014);甘肃省科技重大专项(编号1102FKDA005)

摘　　要：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(human bone mesenchymal stem cells,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、salidroside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、BAX mRNA表达。结果表明,体外成功分离培养hBMMSC;细胞表达CD44、CD29和HLA-ABC,不表达CD34、CD45和HLA-DR;成骨、成脂诱导21 d,茜素红染色可见钙化结节,油红O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48 h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。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.

关 键 词：红景天苷 间充质干细胞 阿糖胞苷 凋亡 

分 类 号：R285.5[医药卫生—中药学]