淫羊藿苷对大鼠骨髓间充质干细胞迁移作用的影响

CJI (Traditional Chinese Medicine) - - Chinese Journal Of Information On Traditional Chin -

张黎声,韩小晶,罗志荣,邵水金,叶晓春,牟芳芳,国海东

上海中医药大学基础医学院,上海 201203

摘要:目的 探讨淫羊藿苷调控大鼠骨髓间充质干细胞( MSC)迁移作用机制。方法 CCK-8 法检测细胞增殖。建立体外细胞缺糖缺氧模型, Hoechst33342 染色观察细胞凋亡。Western blot检测淫羊藿苷处理后MSC表面基质细胞衍生因子 l(SDF-1)受体趋化因子受体 4(CXCR4)的蛋白表达。Transwell 小室跨膜实验观察淫羊藿苷对MSC迁移的作用。结果 0.01、、0.1 1 μmol/L 淫羊藿苷可明显促进MSC 增殖,10 μmol/L 淫羊藿苷抑制MSC增殖。经体外缺糖缺氧处理后, 0.1、1 μmol/L 淫羊藿苷可明显抑制MSC凋亡。淫羊藿苷可显著促进 MSC CXCR4的蛋白表达,同时可提高MSC跨膜迁移的数量。加入CXCR4 拮抗剂 AMD3100 后,各组间细胞迁移数量无明显差异。结论 一定浓度淫羊藿苷可促进 MSC 的增殖、存活和迁移, SDF-1/CXCR4 信号通路参与淫羊藿苷调控MSC迁移的作用。

关键词:淫羊藿苷;骨髓间充质干细胞;存活;迁移;信号通路

DOI: 10.3969/j.issn.1005-5304.2017.02.013

中图分类号: R285.5 文献标识码: A 文章编号: 1005-5304(2017)02-0044-05

Effects of Icariin on Migration of Mesenchymal Stem Cells of Rat Bone Marrow

ZHANG Li-sheng, HAN Xiao-jing, LUO Zhi-rong, SHAO Shui-jin, YE Xiao-chun, MOU Fang-fang, GUO Hai-dong (School of Basic Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China)

Abstract: Objective To explore the mechanism of icariin mediating migration of mesenchymal stem cells (MSC) in rat bone marrow. Methods MSC proliferation was detected by CCK-8 test. Cell apoptosis was examined with Hoechst33342 staining after the establishment of cellular oxygen and glucose deprivation model. The protein expressions of CXCR4, the receptor of SDF-1, in the surface of MSC after stimulated by icariin were detected through Western blot. The migration of MSC was observed by Transwell chemotaxis assay. Results 0.01 μmol/L, 0.1 μmol/L and 1 μmol/L of icariin could significantly promote the proliferation of MSC, while 10 μmol/L of icariin inhibited the proliferation of MSC. After treatment of oxygen and glucose deprivation in vitro, 0.1 μmol/L and 1 μmol/L of icariin could inhibit the apoptosis of MSC. Icariin could not only improve the expression of CXCR4 in MSC, but also increase the number of transmembrane migrated MSC. After the addition of CXCR4 antagonist AMD3100, there was no significant difference in the number of cell migration among the different groups. Conclusion Icariin with appropriate concentration can promote the proliferation, survival and migration of MSC. SDF-1/CXCR4 signaling pathway is involved in the regulation of MSC migration by icariin.

Key words: icariin; mesenchymal stem cell; survival; migration; signaling pathway

干细胞是一类具有自我更新和多向分化潜能的原始细胞。在一定条件下,干细胞可以分化成多种功能细胞,成为治疗损伤性疾病极具潜力的疗法。在心

基金项目:

国家自然科学基金(81673729);上海市自然科学

基金(16ZR1437300);上海市教育委员会科研创新项目(14YZ053)

国海东,E-mail:hdguo8@hotmail.com

通讯作者:

血管领域,近年来的临床研究表明,干细胞移植在一

[1]定程度上有利于心功能的恢复 。我们的研究发现,骨髓间充质干细胞( mesenchymal stem cell,MSC)移植可降低大鼠心肌梗死后梗死面积,促进血管新生

[2-3]

和旁分泌 。然而,心肌梗死后局部微环境不利影

[4]响干细胞的存活,也限制了干细胞的迁移 。淫羊藿苷( icariin)是一种从淫羊藿中提取的异黄酮类化合

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