HIF－1信号通路在介导DMOG动员MSCs中的作用

研究报告

HIF-1信号通路在介导DMOG动员MSCs中的作用

胡韶君1，余勤1，刘丽珍2，葛婷婷1

(1．浙江中医药大学生命科学学院，杭州310053；

2．浙江大学医学院附属第一医院骨髓移植中心，杭州310003)

【摘要】目的探讨HIF-1及下游SDF-1α/CXCR4和VEGF/VEGFR 通路在介导DMOG动员MSCs中的作用机制。 方法 将雄性SD大鼠，随机分为五组：生理盐水对照组、DMOG组、YC-1组、AMD3100组、SU5416组。分别采用CFU-F法与流式细胞术检测大鼠骨髓和外周血中MSCs的数量；ELISA法检测骨髓上清及外周血清中SDF-1α及VEGF蛋白浓度；Western blotting法检测骨髓细胞中HIF-1α、SDF-1α及VEGF蛋白表达水平。 结果 同NS组比较，DMOG组CFU-Fs 数量显著增加，且CD45-CD90+细胞群比例增加(P < 0.05)；同DMOG组比较，YC-1组、AMD3100组、SU5416组CFU-Fs数量均显著减少，且CD45-CD90+细胞群比例降低(P < 0.05)；同DMOG组比较，YC-1组HIF-1α浓度及蛋白表达显著降低(P < 0.05)，AMD3100组SDF-1α浓度及蛋白表达显著降低(P < 0.05)，SU5416组VEGF浓度及蛋白表达显著降低(P < 0.05)。 结论 DMOG可能通过上调HIF-1α，从而调控其下游SDF-1α/CXCR4通路与VEGF/VEGFR通路诱导MSCs 的动员。

【关键词】间充质干细胞动员；DMOG；缺氧诱导因子-1α；基质细胞衍生因子-lα；血管内皮生长因子

[基金项目]国家自然科学基金(81270566)；教育部高校博士点专项科研基金(20130101120022)。

[作者简介]胡韶君(1989-)，硕士，研究方向：干细胞基础与临床研究。

[通讯作者]余勤(1962-)，教授，博士生导师，研究方向：干细胞基础与临床研究，E-mail: qinyu3587@aliyun.com。

【中图分类号】R332【文献标识码】 A

doi:10.3969.j.issn.1671.7856. 2015.001.002

Mechanism of HIF-1 signaling pathway in mediating

MSCs mobilization with DMOG

HU Shao-jun1，YU Qin1，LIU Li-zhen2，GE Ting-ting1

(1. College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China;

2. Bone Marrow Transplantation Center, The First Affiliate Hospital, Medical School of Zhejiang University,

Hangzhou 310003, China)

Abstract【】ObjectiveTo explore the role of HIF-1 and its downstream SDF-1α/CXCR4 and VEGF/VEGFR pathway in mediating MSC mobilization with DMOG. Methods Male SD rats were randomly divided into five groups: Normal saline control group, DMOG group, YC-1 group, AMD3100 group, SU5416 group. We used CFU-F assay and flow cytometry to determine the number of MSCs in rat bone marrow (BM) and peripheral blood (PB) in each group, respectively. The concentrations of SDF-1α and VEGF both in BM and PB serum in each group were detected by ELISA. Western blotting was used to test protein levels of HIF-1α, SDF-1α and VEGF in BM. ResultsCompared with NS group, the number of CFU-Fs as well as the percentage of CD45-CD90+cells increased in DMOG group (P < 0.05); Compared with DMOG group, the number of CFU-Fs as well as the percentage of CD45-CD90+cells decreased in YC-1 group, AMD3100 group and SU5416 group (P < 0.05). Compared with DMOG group, the concentration and protein expression of HIF-1α decreased significantly in YC-1 group (P < 0.05), the concentration and protein expression of SDF-1α decreased significantly in AMD3100 group (P < 0.05), the concentration and protein expression of VEGF decreased significantly in SU5416 group (P < 0.05). Conclusion DMOG can induce MSCs mobilization possibly via up-regulating the expression of HIF-1α and activating its downstream SDF-1α/CXCR4 and VEGF/VEGFR pathway.

【Key words】Mesenchymal stem cells mobilization；DMOG；Hypoxia inducible factor-1α；Stromalcell-derived factor-lα；Vascular endothelial growth factor

间充质干细胞(mesenchymal stem cells，MSCs)动员指应用动员剂或采用动员措施促使骨髓中MSCs释放并迁移到外周血[1]。在组织损伤需要MSCs参与修复的第一时间，给予动员措施，MSCs即可在短时间内从骨髓进入循环池，迁移、归巢入损伤部位，参与损伤修复，此举具有体外细胞移植修复损伤无法比拟的低创性和高效性，因此MSCs动员方法及其机制的研究具有重要的临床意义[1-3]。但是，目前尚缺乏临床可行的MSCs 动员措施，动员机制也不明确。我们前期实验研究发现，脯氨酸羟化酶抑制剂—二甲基乙二酰基甘氨酸(dimethyloxaloylglycine，DMOG)对小鼠MSCs具有动员作用[4]。……