CAJ | 학술논문

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于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索引起[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.
배경:장기복용배포소가억제성골세포분화,도치골질소송,이식물자격소자마분가촉진성골세포증식여분화.목적:탐토자마분대배포소처리적소서골수간충질우세포증식급향성골세포분화적영향,분석기가능존재적신호통로.방법:첩벽법분리배양소서골수간충질간세포,분위5조,분별급여자마분、배포소、자마분+배포소、자마분+배포소+ICI182780、자마분+배포소+L-NAME.도치현미경하관찰골수간충질간세포적형태학변화급광화능력,통과검측[3H]-갑기흉선밀정참입솔반영세포증식정황,통과측정세포내감성린산매활성여개침적량반영세포향성골세포분화능력,측정배양액중일양화담산량여세포내배린산조감함량.결과여결론:합용자마분여배포소가완전역전단용배포색인기[3H]-갑기흉선밀정참입솔、세포내감성린산매활성、개침적량적감소(P<0.05혹0.01).병반수일양화담산량여세포내배린산조감함량적회복(P<0.01),동시도치현미경하가관찰도간예12 d시교고적세포생장밀도화광화결절형성,이상작용가피자격소수체길항제ICI182780、일양화담합매억제제L-NAME취소.제시자마분가통과자격소수체/일양화담/배린산조감신호통로역전배포소억제골수간충질간세포적증식급향성골세포적분화.
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 monophosphata(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.