7-甲氧基-4’-羟基异黄酮促进大鼠成骨细胞成骨性分化的作用机制
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摘要
目的探讨7-甲氧基-4’-羟基异黄酮(MHIF)促进成骨细胞成熟矿化是否与NO/cGMP/sGC信号通路相关。方法检测成骨细胞经不同浓度(0,10~(-4),10~(-5),10~(-6),10~(-7),10~(-8) mol·L~(-1)) MHIF处理后骨钙素和碱性磷酸酶确定最适药物浓度。使用一氧化合成酶阻断剂N-单甲基-L-精氨酸(L-NMA)处理成骨细胞,观察MHIF对骨钙素和碱性磷酸酶影响,一氧化氮(NO)和3`-5`-环鸟苷-磷酸(cGMP)的含量;应用蛋白质印迹检测细胞中蛋白可溶性的鸟氨酸环化酶和蛋白激酶G的表达水平。结果经10~(-6 ) mol·L~(-1) MHIF处理后成骨细胞中碱性磷酸酶活性和骨钙素的含量升高,预先使用L-NMA处理成骨细胞后,MHIF提高成骨细胞中碱性磷酸酶活性和骨钙素含量的作用受到抑制,MHIF提高一氧化氮合酶(NOS)、NO、cGMP、sGC和PKG表达均受到抑制。结论 MHIF可通过NO/cGMP/sGC信号通路促进体外培养成骨细胞成熟与矿化。
Objective To determine whether 7-methoxy-4'-hydroxyisoflavone promote maturation of osteoblasts. Methods Osteoblasts was treated by different concentrations(0, 10~(-4), 10~(-5), 10~(-6), 10~(-7), 10~(-8) mol·L~(-1)) of 7-methoxy-4'-hydroxyisoflavone. The concentration of osteocalcin(OC) and alkaline phosphatase(ALP) were determined.The expression levels of protein sGC and PKG-1 in cells were detected by Western blotting. Results The group of 10~(-6) mol·L~(-1) significantly increased ALP activity and OC content in osteoblasts. The effect of L-NMA on ALP activity and OC content in osteoblasts was inhibited, and the expression of NOS, NO, cGMP, sGC and PKG was inhibited Conclusion 7-methoxy-4'-hydroxyisoflavone promoted osteoblasts differentiation through NO/cGMP/sGC signal pathway.
Objective To determine whether 7-methoxy-4'-hydroxyisoflavone promote maturation of osteoblasts. Methods Osteoblasts was treated by different concentrations(0, 10~(-4), 10~(-5), 10~(-6), 10~(-7), 10~(-8) mol·L~(-1)) of 7-methoxy-4'-hydroxyisoflavone. The concentration of osteocalcin(OC) and alkaline phosphatase(ALP) were determined.The expression levels of protein sGC and PKG-1 in cells were detected by Western blotting. Results The group of 10~(-6) mol·L~(-1) significantly increased ALP activity and OC content in osteoblasts. The effect of L-NMA on ALP activity and OC content in osteoblasts was inhibited, and the expression of NOS, NO, cGMP, sGC and PKG was inhibited Conclusion 7-methoxy-4'-hydroxyisoflavone promoted osteoblasts differentiation through NO/cGMP/sGC signal pathway.
引文
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[2] FILIPOVIC B,SOSIC-JURJEVIC B,AJDZANOVIC V,et al.Daidzein administration positively affects thyroid C cells and bone structure in orchidectomized middle-aged rats [J].Osteoporos Int,2010,21:1609-1616.
[3] STRONG A L,OHLSTEIN J F,JIANG Q,et al.Novel daid-zein analogs enhance osteogenic activity of bone marrow-derived mesenchymal stem cells and adipose-derived stromal /stem cells through estrogen receptor dependent and independent mechanisms [J].Stem Cell Res Ther,2014,54(4):105.
[4] MA H P,MING LG,GE B F,et al.Icariin is more poten tthan genisteinin promoting osteoblast differentiation and mineralization in vitro[J].J Cell Biochem,2011,112(3):916-923.
[5] 王威,石元刚,尹黎明,等.苦瓜素大豆异黄酮及其复合物对高脂大鼠血脂和血浆一氧化氮的影响[J].第三军医大学学报,2005,27(10):1006-1008.
[6] 吴江群.骨髓基质干细胞体内成骨分化研究[D].北京:中国协和医科大学,2008:10-39.
[7] KALYANARAMAN H,SCHALL N,PILZ R B.Nitric oxide and cyclic GMP functions in bone[J].Nitric Oxide,2018,76:62-70.
[8] CAPULLI M,OLSTAD O K,ONNERFJORD P,et al.The C-terminal domain of chondroadherin:a new regulator of osteoclast motility counteracting bone loss[J].J Bone Miner Res,2014,29(8):1833-1846.
[9] FERRINI M G,GARCIA E,ABRAHAM A,et al.Effect of ginger,Paullinia cupana,muira puama and L-citrulline,singly or in combination,on modulation of the inducible nitric oxide- NO-cGMP pathway in rat penile smooth muscle cells[J].Nitric Oxide,2018,76:81-86.
[10] RIANCHO J A,SALAS E,ZARRABEITIA M T,et al.Expression and functional role of nitric oxide synthase in osteoblast-like cells[J].J Bone Miner Res,1995,10(3):439-446.
[11] MANCINI L,MORADI-BIDHENDI N,BECHERINI L,et al.The biphasic effects of nitric oxide in primary rat osteoblasts are cGMP dependent[J].Biochem Biophys Res Commun,2000,274(2):477-481.
[12] 康小龙,田红林,何承辉,等.刺山柑总生物碱对系统性硬皮病转化生长因子-β1/Smad4通路的调控作用[J].医药导报,2018,37 (11):1307-1310.