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医药导报  2019, Vol. 38 Issue (6): 730-734    DOI: 10.3870/j.issn.1004-0781.2019.06.009
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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*
宋明甲1(),文益民1,吴晓燕1,柴晓亮1,周建2()
1.中国人民解放军联勤保障部队九四〇医院 脊柱外科,兰州 730050
2.中国人民解放军联勤保障部队九四〇医院 骨科研究所,兰州 730050
Mechanism of 7-Methoxy-4’-hydroxyisoflavone on the Promotion of OsteogenesIs Differentiation of Rat Osteoblast Cultured
Mingjia SONG1(),Yiming WEN1,Xiaoyan WU1,Xiaoliang CHAI1,Jian ZHOU2()
1.Spinal Surgery, Joint Logistics Support Force 940 Hospital, CPLA, Lanzhou 730050, China
2.Institute of Orthopaedics, Joint Logistics Support Force 940 Hospital, CPLA, Lanzhou 730050, China
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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。

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宋明甲
文益民
吴晓燕
柴晓亮
周建
关键词 7-甲氧基-4'-羟基异黄酮成骨细胞骨钙素N-单甲基-L-精氨酸    
Abstract

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.

Key words7-Methoxy-4'-hydroxyisoflavone    Osteoblast    Osteocalcin    N-monomethyl-L-arginine
收稿日期: 2018-04-27      出版日期: 2019-06-11
基金资助:*甘肃省自然科学基金资助项目(1506RJZA306)
引用本文:   
宋明甲,文益民,吴晓燕,柴晓亮,周建. 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*[J]. 医药导报, 2019, 38(6): 730-734.
Mingjia SONG,Yiming WEN,Xiaoyan WU,Xiaoliang CHAI,Jian ZHOU. Mechanism of 7-Methoxy-4’-hydroxyisoflavone on the Promotion of OsteogenesIs Differentiation of Rat Osteoblast Cultured. Herald of Medicine, 2019, 38(6): 730-734.
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http://www.yydbzz.com/CN/10.3870/j.issn.1004-0781.2019.06.009      或      http://www.yydbzz.com/CN/Y2019/V38/I6/730
图1  不同浓度7-甲氧基-4’-羟基异黄酮对作用不同时间成骨细胞后ALP活性($\bar{x}±s$,n=3)
图2  不同浓度7-甲氧基-4’-羟基异黄酮处理成骨细胞后骨钙素含量($\bar{x}±s$,n=3)
图3  L-NMA对MHIF提高ALP活性的影响($\bar{x}±s$,n=3)
与对照组比较,*1P<0.01;与MHIF组比较,*2P<0.05
图4  L-NMA对MHIF提高骨钙素的影响($\bar{x}±s$,n=3)
与对照组比较,*1P<0.01;与MHIF组比较,*2P<0.01。。
图5  L-NMA对MHIF提高NO含量的影响($\bar{x}±s$,n=3)
与对照组比较,*1P<0.01;与MHIF组比较,*2P<0.05
图6  L-NMA对MHIF提高cGMP含量的影响($\bar{x}±s$,n=3)
与对照组比较,*1P<0.01;与MHIF组比较,与*2P<0.05
图7  MHIF对体外培养成骨细胞内sGC和PKG蛋白表达水平的影响
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