基于斑马鱼模型和代谢组学技术的西洋参抗骨质疏松作用机制研究  

Anti-osteoporosis mechanism of Panax quiquefolium L.based on zebrafish model and metabonomics

作　　者：邱钺姿 王传森 徐凤华 张轩铭 王利振 李培海 刘可春 屠鹏飞[4] 林厚文[5] 张姗姗 李晓彬 QIU Yue-zi;WANG Chuan-sen;XU Feng-hua;ZHANG Xuan-ming;WANG Li-zhen;LI Pei-hai;LIU Ke-chun;TU Peng-fei;LIN Hou-wen;ZHANG Shan-shan;LI Xiao-bin(Biology Institute,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250103,China;Engineering Research Center of Zebrafish Models for Human Diseases and Drug Screening of Shandong Province,Jinan 250103,China;School of Basic Medical Sciences,Qingdao University,Qingdao 266000,China;School of Pharmaceutical Sciences,Peking University,Beijing 100191,China;Ren Ji Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 200127,China)

机构地区：[1]齐鲁工业大学(山东省科学院),山东省科学院生物研究所,山东济南250103 [2]山东省人类疾病斑马鱼模型与药物筛选工程技术研究中心,山东济南250103 [3]青岛大学基础医学院,山东青岛266000 [4]北京大学药学院,北京100191 [5]上海交通大学医学院附属仁济医院,上海200127

出　　处：《药学学报》2023年第7期1894-1903,共10页Acta Pharmaceutica Sinica

基　　金：山东省重点研发计划(重大科技创新工程)(2021CXGC010507,2022CXGC020515);国家自然青年科学基金资助项目(42006090);济南市高校20条资助项目(2020GXRC031);泰山学者特聘专家项目(ts20190950);生物基材料与绿色造纸国家重点实验室开放基金资助项目(ZZ20190413);山东省自然科学基金面上项目(ZR2022MD102);齐鲁工业大学(山东省科学院)科教产融合试点工程重大创新专项(2022JBZ01-06);齐鲁工业大学(山东省科学院)科教产融合试点工程基础研究类项目(2022PX061).

摘　　要：本研究基于斑马鱼模型结合代谢组学技术探究西洋参提取物抗骨质疏松活性及其作用机制。采用泼尼松龙诱导的骨质疏松斑马鱼模型,以骨骼荧光面积和荧光密度为评价指标,开展西洋参的抗骨质疏松活性研究,实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测西洋参对斑马鱼成骨细胞相关基因及破骨细胞相关基因表达的影响,基于超高效液相色谱-质谱联用法(ultra-high performance liquid chromatography-mass spectrometry,UPLC-MS)的代谢组学技术探讨其生物标志物的变化规律和影响的代谢通路。结果表明,与模型组相比,中国吉林、加拿大、中国文登、美国产的西洋参50%乙醇提取物均可显著提高斑马鱼骨骼荧光面积,除美国产西洋参外,其余产地西洋参50%乙醇提取物均可显著提高斑马鱼骨骼荧光密度。PCR结果表明,西洋参可显著上调vitamin D receptor b(vdrb)、collagen type Iα2(col1a2)、cysteine-rich acidic secreted protein(sparc),下调matrix metalloproteinase9(mmp9)、anti-tartrase acid phosphatase(trap)、cathepsin K(ctsk)基因的表达。代谢组学分析鉴定出24个关键差异代谢物,通路分析表明西洋参可通过参与嘌呤代谢、三羧酸循环、磷酸戊糖代谢途径,回调10个关键生物质量标志物含量,从而改善斑马鱼骨质疏松状态。本研究初步揭示了西洋参50%醇提物通过多靶点、多通路发挥抗骨质疏松活性,为西洋参抗骨质疏松产品的开发利用提供理论基础。本实验获得山东省科学院生物研究所实验动物福利伦理委员会批准(批准号:SWS20181002)。In this study,we investigated the anti-osteoporotic activity and mechanism of action of extract of Panax quiquefolium L.based on zebrafish model combined with metabolomics technology.A zebrafish model of prednisolone-induced osteoporosis was used to compare the anti-osteoporotic activity of Panax quiquefolium L.,and the expression of osteoblast-associated genes and osteoclast-associated genes in zebrafish was detected by quantitative real-time PCR(qRT-PCR),using bone fluorescence area and fluorescence density as evaluation indexes.Metabolomics based on ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS)was used to explore the change patterns of biomarkers and the metabolic pathways affected.The results showed that the 50%ethanol extracts of Panax quiquefolium L.from Jilin,Canada,Wenden and the United States can significantly improve the bone fluorescence area of zebrafish compared with model group.Furthermore,four sources 50%ethanol extracts of Panax quiquefolium L.except United States also can significantly improve the bone fluorescence density of zebrafish.In addition,PCR showed that extract of Panax quiquefolium L.can significantly up-regulated the expression of vitamin D receptor b(vdrb),collagen type Iα2(col1a2)and cysteine-rich acidic secreted protein(sparc)genes,and down-regulated the expression of matrix metalloproteinase 9(mmp9),anti-tartrase acid phosphatase(trap)and cathepsin K(ctsk)genes.Metabolomic analysis identified 24 key differential metabolites.Furthermore,pathway analysis showed that Panax quiquefolium L.could regulate the levels of 10 key biomarkers by participating in purine metabolism,tricarboxylic acid cycle and pentose phosphate metabolism and improve the osteoporosis status of zebrafish.This study preliminically revealed the anti-osteoporosis mechanism of 50%ethanol extract from Panax quiquefolium L.through multi-component,multi-target and multi-pathway and also provides theoretical basis for clinical development and utilization of anti-osteoporosis products of Panax

关 键 词：西洋参 代谢组学 斑马鱼 骨质疏松 作用机制 

分 类 号：R917[医药卫生—药物分析学]