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| 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 |
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| 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） |
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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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