骨碎补-续断药对对成骨/破骨代谢的双向调控作用及其对Hif1ɑ基因的影响
Effects of rhizoma drynariae-radix dipsaci herb pair on osteogenesis/osteoclastogenesis and Hif1ɑGene
  
DOI:10.3969/j.issn.1006-7108.2023.01.012
中文关键词:  中医中药  骨质疏松  成骨细胞  破骨细胞  骨碎补  续断  Hif1ɑ
英文关键词:traditional Chinese medicine  osteoporosis  osteogenesis  osteoclastogenesis  rhizoma drynariae  radix dipsaci  Hif1ɑ
基金项目:福建省科技厅省属公益类科研院所基本科研专项(2020R1003007);福建省科技厅省自然科学基金项目(2021J01917)
作者单位
陈玄1,2 陈娟1,2 谢丽华1,2 李生强1,2 黄景文1,2 叶云金1,2 陈赛楠1,2 黄小彬1,2 葛继荣1,2* 1.福建省中医药科学院骨质疏松证候基因组学重点研究室,福建 福州350003 2.福建省中西医结合防治骨质疏松重点实验室,福建省中医药科学院及福建中医药大学附属康复医院,福建 福州 350003 
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中文摘要:
      目的 分析骨碎补-续断药对对成骨/破骨代谢的调控作用并初步探讨其机制。方法 分别制备空白、低剂量、中剂量及高剂量的骨碎补-续断含药血清。采用CCK-8法、ALP染色法和茜素红染色法,分别观察含药血清对MC3T3-E1细胞增殖、成骨和矿化能力的影响;采用CCK-8法和TRAP染色法观察含药血清对RAW264.7细胞增殖和破骨分化能力的影响。采用系统药理学的方法分析药物的可能作用靶点,并结合RT-PCR和Western-blot的方法验证。结果 中剂量和高剂量骨碎补-续断含药血清可促进MC3T3-E1细胞的增殖,抑制RAW264.7细胞的增殖,中剂量组和高剂量组间没有明显差异;同时,中、高剂量含药血清可促进MC3T3-E1细胞的ALP活性和钙化能力,抑制RAW264.7细胞的TRAP活性,中剂量和高剂量间没有显著性差异。骨碎补-续断药对的可能作用靶点涉及HIF1ɑ、RT-PCR和Western-blot,结果证实中剂量含药血清可提高MC3T3-1细胞Hif1ɑ基因的mRNA和蛋白水平及RAW264.7细胞的HIF1ɑ蛋白水平。结论 骨碎补-续断药对具有促进成骨代谢,抑制破骨代谢的作用,HIF1ɑ可能是骨碎补-续断药对的一个重要作用靶点。
英文摘要:
      Objective To analyze the regulation effects of rhizoma drynariae-radix dipsaci herb pair on osteogenesis/osteoclastogenesis and to obtain insight into its molecular mechanism. Methods Low, medium, and high doss of serum containing herb pair rhizoma drynariae-radix dipsaci and the control were prepared, respectively. The effects of the herb pair on proliferation, osteogenesis, and mineralization of MC3T3-E1 cells were observed with CCK-8 staining, ALP staining, and Alizarin red staining, respectively. The effects on proliferation and osteoclastogenesis of RAW264.7 cells were observed with CCK-8 staining and TRAP staining. The possible targets of the herbs were analyzed with systematic pharmacology and confirmed using RT-PCR and Western blotting. Results Serum containing the herb pair promoted the proliferation of MC3T3-E1 cells and inhibited the proliferation of RAW264.7 cells in a dose-dependent manner. Furthermore, medium and high doses of drug serum promoted the ALP activity and calcification ability of MC3T3-E1 cells, but inhibited the TRAP activity of RAW264.7 cells. One of the possible targets of rhizoma drynariae-radix dipsaci herb pair was HIF1ɑ. RT-PCR and Western blotting results confirmed that medium dose of drug containing serum raised mRNA and protein levels of Hif1ɑ gene in MC3T3-1 cell and protein level of HIF1ɑ in RAW264.7 cells. Conclusion Rhizoma drynariae-radix dipsaci herb pair promotes osteogenesis and inhibits osteoclastogenesis. HIF1ɑ may be a key target of the herb pair.
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