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。
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 result confirmed that medium dose of drug containing serum raised mRNA and protein levels of Hif1ɑgene in MC3T3-1 cell and protein level of HIF1ɑin RAW264.7 cells.Conclusion Rhizoma drynariae-radix dipsaci herb pair promotes osteogenesis and inhibits osteoclastogenesis.HIF1ɑmay be a key target of the herb pair.
作者
陈玄
陈娟
谢丽华
李生强
黄景文
叶云金
陈赛楠
黄小彬
葛继荣
CHEN Xuan;CHEN Juan;XIE Lihua;LI Shengqiang;HUANG Jingwen;YE Yunjin;CHEN Sainan;HUANG Xiaobin;GE Jirong(The Key Research Laboratory of Osteoporosis Syndrome Genomics,Fujian Academy of Chinese Medical Sciences,Fuzhou 350003;Fujian Key Laboratory of Integrated Medicine for Prevention and Treatment of Osteoporosis,Fujian Academy of Chinese Medical Sciences&Rehabilitation Hospital affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou 350003,China)
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2023年第1期64-69,共6页
Chinese Journal of Osteoporosis
基金
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)。
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
traditional Chinese medicine
osteoporosis
osteogenesis
osteoclastogenesis
rhizoma drynariae
radix dipsaci
Hif1ɑ
