右归丸入血成分对Wnt/β-catenin、OPG/RANKL/RANK通路相关因子mRNA表达的影响
Effects of blood components of Yougui Wan on mRNA expressions of Wnt/β-catenin and OPG/RANKL/RANK pathway related factors
  
DOI:10.3969/j.issn.1006-7108.2025.08.009
中文关键词:  右归丸入血成分  MC3T3-E1  RAW264.7  Wnt/β-catenin  OPG/RANKL/RANK  信号通路  mRNA  骨质疏松
英文关键词:blood components of Yougui Wan  MC3T3-E1  RAW264.7  Wnt/β-catenin  OPG/RANKL/RANK  signaling pathway  mRNA  osteoporosis
基金项目:国家自然科学基金面上项目(81973868)
作者单位
商佳璐1 叶怡馨1 王娅红1 章建华2 尹华1* 1.浙江中医药大学药学院中药标准化研究实验室浙江 杭州 311402 2.浙江中医药大学附属第一医院浙江 杭州 310006 
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中文摘要:
      目的 探讨右归丸入血成分对MC3T3-E1和RAW264.7细胞Wnt/β-catenin、OPG/RANKL/RANK通路相关因子mRNA表达的影响。方法 用MC3T3-E1和RAW264.7细胞分别进行成骨和破骨的诱导培养,制备右归丸入血成分的单体干预液进行细胞干预,通过RT-qPCR检测MC3T3-E1细胞中Lrp6、Lrp5、β-catenin、Tcf7、Fzd2、OPG及RAW264.7细胞中Traf6、Nfatc1、Fos、Mmp9、Ctsk、Car2 mRNA的表达。结果 与空白对照组相比,乌头碱能显著抑制Traf6、Fos和Mmp9的mRNA表达(P<0.05);次乌头碱和新乌头碱能显著抑制Fos的mRNA表达(P<0.05);苯甲酰乌头原碱可显著上调Lrp6、β-catenin、Tcf7、Fzd2和OPG的mRNA表达(P<0.05),显著下调Traf6、Fos和Mmp9的mRNA(P<0.05);苯甲酰次乌头原碱可促进Lrp5、β-catenin、Tcf7、Fzd2和OPG的mRNA表达(P<0.05);苯甲酰新乌头原碱可促进Lrp6、Lrp5、Tcf7和OPG的mRNA表达(P<0.05),抑制Traf6、Fos、Mmp9和Car2的表达(P<0.05);松脂醇二葡萄糖苷可促进Lrp5、β-catenin、Tcf7和OPG的mRNA表达(P<0.05),显著下调Traf6、Ctsk、Car2的mRNA表达(P<0.05);肉桂酸能显著上调Lrp5、Tcf7、Fzd2和OPG的mRNA表达(P<0.05),显著抑制Nfatc1和Fos的mRNA表达(P<0.05)。结论 右归丸入血成分在一定浓度范围内可以通过调控Wnt/β-catenin、OPG/RANKL/RANK信号通路相关因子的基因表达影响骨形成和骨吸收,起到治疗骨质疏松的作用。
英文摘要:
      Objective To clarify the effects of blood components of Yougui Wan on the mRNA expressions of Wnt/β-catenin and OPG/RANKL/RANK pathway related factors in MC3T3-E1 and RAW264.7 cells. Methods MC3T3-E1 and RAW264.7 cells were cultured for osteogenic and osteoclast induction, respectively. Monomer intervention solution containing blood components of Yougui Wan was prepared for cell intervention. RT qPCR was used to detect the expressions of Lrp6, Lrp5, β - catenin, Tcf7, Fzd2, and OPG in MC3T3-E1 cells and Traf6, Nfatc1, Fos, Mmp9, Ctsk, and Car2 mRNA in RAW264.7 cells. Results The staining showed that MC3T3-E1 cells were blue-purple with orange-red calcium deposits, and RAW264.7 cells were burgundy. Compared to that in the blank control group, aconitine significantly inhibited the mRNA expressions of Traf6, Fos, and Mmp9 (P<0.05). Subaconitine and neoaconitine significantly inhibited the mRNA expression of Fos (P<0.05). Benzoyl aconitine significantly up-regulated the mRNA expressions of Lrp6, β-catenin, Tcf7, Fzd2, and OPG (P<0.05), and significantly down-regulated the mRNA expressions of Traf6, Fos, and Mmp9 (P<0.05). Benzoyl aconitine promoted mRNA expressions of Lrp5, β-catenin, Tcf7, Fzd2, and OPG (P<0.05). Benzoyl neoaconitine promoted mRNA expressions of Lrp6, Lrp5, Tcf7, and OPG (P<0.05), and inhibited the expressions of Traf6, Fos, Mmp9, and Car2 (P<0.05). Pineolin glucoside promoted the mRNA expressions of Lrp5, β-catenin, Tcf7, and OPG (P<0.05), and significantly down-regulated the mRNA expressions of Traf6, Ctsk, and Car2 (P<0.05). Cinnamic acid significantly up-regulated the mRNA expressions of Lrp5, Tcf7, Fzd2, and OPG (P<0.05), and significantly inhibited the mRNA expressions of Nfatc1 and Fos (P<0.05). Conclusion The blood components of Yougui Wan regulates the gene expressions of Wnt/β-catenin and OPG/RANKL/RANK signaling pathway related factors within a certain concentration range, affects bone formation and resorption, and plays a role in the treatment of osteoporosis.
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