中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (36): 6833-6836.doi: 10.3969/j.issn.1673-8225.2010.36.042

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

雌激素受体/一氧化氮/环磷酸鸟苷通路介导雌马酚逆转环孢素抑制骨髓间充质干细胞增殖及向成骨细胞的分化

宋丽华1，黄  燕2，武翠玲2，石变华2   

  1. 长治医学院，1药理学教研室，2心血管研究所，山西省长治市  046000
  • 出版日期:2010-09-03 发布日期:2010-09-03
  • 作者简介:宋丽华★，女，1970年生，辽宁省盖州市人，汉族，2003年中南大学湘雅医学院毕业，硕士，副教授，主要从事植物雌激素与干细胞定向分化方面的研究。

Equol reverses the inhibition of cyclosporin A on the proliferation and osteoblastic differentiation of bone marrow mesenchymal stem cells through estrogen receptor/ nitric oxide / cyclic guanosine monophosphate signal pathway

Song Li-hua1, Huang Yan2, Wu Cui-ling2, Shi Bian-hua2   

  1. 1 Department of Pharmacology, 2 Institute of Cardiovascular Disease, Changzhi Medical College, Changzhi  046000, Shanxi Province, China
  • Online:2010-09-03 Published:2010-09-03
  • About author:Song Li-hua★, Master, Associate professor, Department of Pharmacology, Changzhi Medical College, Changzhi 046000, Shanxi Province, China slh10282001@yahoo.com.cn

PDF

340

被引次数

2

摘要：

背景：长期服用环孢素可抑制成骨细胞分化，导致骨质疏松，而植物雌激素雌马酚可促进成骨细胞增殖与分化。
目的：探讨雌马酚对环孢素处理的小鼠骨髓间充质干细胞增殖及向成骨细胞分化的影响，分析其可能存在的信号通路。
方法：贴壁法分离培养小鼠骨髓间充质干细胞，分为5组，分别给予雌马酚、环孢素、雌马酚+环孢素、雌马酚+环孢素+ICI182780、雌马酚+环孢素+L-NAME。倒置显微镜下观察骨髓间充质干细胞的形态学变化及矿化能力，通过检测[3H]-甲基胸腺嘧啶掺入率反映细胞增殖情况，通过测定细胞内碱性磷酸酶活性与钙沉积量反映细胞向成骨细胞分化能力，测定培养液中一氧化氮产量与细胞内环磷酸鸟苷含量。
结果与结论：合用雌马酚与环孢素可完全逆转单用环孢素引起[3H]-甲基胸腺嘧啶掺入率、细胞内碱性磷酸酶活性、钙沉积量的减少(P < 0.05或0.01)，并伴随一氧化氮产量与细胞内环磷酸鸟苷含量的恢复(P < 0.01)，同时倒置显微镜下可观察到干预12 d时较高的细胞生长密度和矿化结节形成，以上作用可被雌激素受体拮抗剂ICI182780、一氧化氮合酶抑制剂L-NAME取消。提示雌马酚可通过雌激素受体/一氧化氮/环磷酸鸟苷信号通路逆转环孢素抑制骨髓间充质干细胞的增殖及向成骨细胞的分化。

关键词: 雌马酚, 环孢素, 骨髓间充质干细胞, 成骨细胞分化, 雌激素受体, 一氧化氮, 环磷酸鸟苷

Abstract:

BACKGROUND: Long term taking cyclosporin A (CsA) can inhibit osteoblastic differentiation and induce osteoporosis. Equol, which has greater binding affinity to estrogen receptors, can stimulate the proliferation and osteoblastic differentiation.
OBJECTIVE: To investigate whether equol may protect against the proliferation and osteoblastic differentiation inhibited by CsA in mouse bone marrow mesenchymal stem cells (BMSCs) cultures and analyze signal pathway of protection.
METHODS: Primary mouse BMSCs were cultured by using attachment method and assigned to five groups, which respectively treated with equol or/and CsA in the presence or absence of ICI182780, an estrogen receptor antagonist, and Nω-nitro-L-arginine methyl ester. Under an inverted microscope, morphological changes and mineralization ability of BMSCs were observed. The cell proliferation was measured by [3H]-thymidine incorporation. The osteoblastic differentiation and mineralization of extracellular matrix in BMSCs was assessed by measuring alkaline phosphatase activity and calcium deposition, respectively. Nitric oxide production in the conditioned media and cyclic guanosine monophosphate (cGMP) content in BMSCs were determined by using commercial nitric oxide and cGMP kit, respectively.
RESULTS AND CONCLUSION: Equol reversed the decreased [3H] thymidine incorporation (P < 0.05), alkaline phosphatase activity (P < 0.05) and calcium deposition (P < 0.01) of CsA, which was accompanied with the changes of nitric oxide production (P < 0.01) and cGMP content (P < 0.01). The group by co-treatment with equol and CsA possessed higher cells growth density and small mineralized nodes than CsA group on day 12 under an inverted microscope. Moreover, the equol-reversed effect was abolished by ICI182780 and Nω-nitro-L-arginine methyl ester. These indicated that equol can reverse the inhibition of CsA on the proliferation and osteoblastic differentiation of mouse BMSCs through estrogen receptor/ nitric oxide/cGMP signal pathway.

中图分类号: