中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (29): 4643-4650.doi: 10.12307/2022.904

• 骨组织构建 bone tissue construction • 上一篇    下一篇

补肾健脾活血方与靶点密切相关组蛋白去甲基化酶JMJD2B在骨质疏松症中促成骨分化：体外细胞实验验证

罗  臻1，黄禹僖1，柴生颋1，2，李飞龙1，2，陈群群1，2   

  1. 1广州中医药大学，广东省广州市  510006；2广州中医药大学第三附属医院，广东省广州市  510378
  • 收稿日期:2021-11-16 接受日期:2021-12-22 出版日期:2022-10-18 发布日期:2022-03-26
  • 通讯作者: 陈群群，博士，副主任医师，广州中医药大学第三附属医院，广东省广州市 510378
  • 作者简介:罗臻，男，1994年生，河南省潢川县人，汉族，2021年广州中医药大学毕业，硕士，主要从事中医药治疗骨与关节疾病方面的研究。
  • 基金资助:
    广东省自然科学基金博士启动纵向协同项目(2018A030310606)，项目负责人：陈群群；广东省中医药管理局科研项目(20202085)，项目负责人：李飞龙

Bushen Jianpi Huoxue Recipe is closely related to the target of histone demethylase JMJD2B in promoting osteogenic differentiation in osteoporosis: an in vitro cell experimental verification

Luo Zhen1, Huang Yuxi1, Chai Shengting1, 2, Li Feilong1, 2, Chen Qunqun1, 2   

  1. 1Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China; 2The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510378, Guangdong Province, China
  • Received:2021-11-16 Accepted:2021-12-22 Online:2022-10-18 Published:2022-03-26
  • Contact: Chen Qunqun, MD, Associate chief physician, Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China; The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510378, Guangdong Province, China
  • About author:Luo Zhen, Master, Guangzhou University of Chinese Medicine, Guangzhou 510006, Guangdong Province, China
  • Supported by:
    the Natural Science Foundation of Guangdong Province for Doctor Collaboration Project, No. 2018A030310606 (to CQQ); the Scientific Research Project of Guangdong Provincial Administration of Traditional Chinese Medicine, No. 20202085 (to LFL)

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摘要：

文题释义：
网络药理学：融合系统生物学、生物信息学、网络科学等学科，从系统层次和生物网络的整体角度出发，解析药物与治疗对象之间的分子关联，揭示药物的系统性药理机制，从而指导新药研发和临床诊疗，是人工智能和大数据时代药物系统性研究的新兴原创学科。
组蛋白修饰：指在保持原有DNA序列的条件下，通过组蛋白的甲基化(Me)、乙酰化(Ac)和磷酸化(P)等修饰以调节DNA转录表达的分子作用。

背景：补肾健脾活血方是临床常用于治疗骨质疏松症的经验用方，但其潜在的分子作用机制尚需深入探讨。
目的：通过网络药理学方法探讨补肾健脾活血方治疗骨质疏松症的潜在分子机制，并初步验证其通过组蛋白甲基化修饰发挥作用的可能性。
方法：通过网络药理学的方法，利用相关数据库及软件筛选出补肾健脾活血方的活性药物成分及作用靶点，获取骨质疏松症相关疾病靶点，取交集得到补肾健脾活血方治疗骨质疏松症的潜在作用靶点。使用String和Cytoscape软件构建药物成分-靶点网络，筛选出核心作用靶点。于GO生物学分析结果提取组蛋白修饰相关功能及相关基因，从核心作用靶点中找到与组蛋白修饰相关靶点基因。结合文献分析靶点基因在骨质疏松症与组蛋白修饰中可能发挥的生物学作用，采用体外细胞实验验证补肾健脾活血方影响与靶点密切相关的组蛋白去甲基化酶JMJD2B在骨质疏松症中可能发挥的促成骨分化作用。
结果与结论：①共筛选出补肾健脾活血方治疗骨质疏松症相关活性药物成分118个，对应靶点165个，核心靶点为IL6、TP53、FOS、JUN、STAT3、RELA、CCND1、MYC、MAPK1、MAPK8、MAPK14、VEGFA、AKT1、ESR1、TNF、NR3C1等；②GO生物学功能分析得出，对药物的反应、对营养水平的反应、对氧化应激的反应、对类固醇激素的反应、转录调节、转录因子结合、磷酸酶结合、蛋白酶结合、类固醇激素受体活性等生物学功能与共同靶点密切相关；③MAPK8、VEGFA和TP53等核心靶点与组蛋白修饰功能有关，木犀草素、槲皮素和山奈酚等可能是影响组蛋白修饰功能治疗骨质疏松症的主要药物成分；④体外细胞实验结果表明，补肾健脾活血方含药血清干预下大鼠骨髓间充质干细胞的成骨相关碱性磷酸酶和JMJD2B、RUNX2蛋白表达均升高(P < 0.05)；⑤与补肾健脾活血方网络药理学靶点密切相关的组蛋白去甲基化酶JMJD2B可能发挥着促成骨分化作用以治疗骨质疏松症。

https://orcid.org/0000-0003-1918-0171 (罗臻) 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 组蛋白去甲基化酶, 骨质疏松症, JMJD2B, 成骨分化, 网络药理学

Abstract: BACKGROUND: Bushen Jianpi Huoxue Recipe is an empirical prescription commonly used in the treatment of osteoporosis. However, its potential molecular mechanism needs to be further explored.
OBJECTIVE: To explore the potential molecular mechanism of Bushen Jianpi Huoxue Recipe in the treatment of osteoporosis based on network pharmacology, and to verify the possibility of its effect through histone methylation modification.
METHODS: Based on the network pharmacology, the active drug components and action targets of Bushen Jianpi Huoxue Recipe were screened from relevant database using relevant software, and the targets of osteoporosis-related diseases were obtained. The intersected targets were obtained and identified as the potential targets of Bushen Jianpi Huoxue Recipe in the treatment of osteoporosis. The drug component-target network was constructed by String and Cytoscape software, and the core targets were screened out. The functions and genes related to histone modification were extracted based on the results of GO biological analysis, and the target genes related to histone modification were screened from the core targets. Literature analysis was performed to indicate the possible biological role of target genes in osteoporosis and histone modification. In vitro cell experiments were conducted to verify the effect of Bushen Jianpi Huoxue Recipe on osteogenic differentiation in the treatment of osteoporosis via the regulation of histone demethylase JMJD2B that is closely related to the target.
RESULTS AND CONCLUSION: A total of 118 active drug components related to osteoporosis were identified, with 165 corresponding targets. The core targets were IL6, TP53, FOS, JUN, STAT3, RELA, CCND1, MYC, MAPK1, MAPK8, MAPK14, VEGFA, AKT1, ESR1, TNF, NR3C1 and so on. GO biological function analysis showed that the biological functions such as drug response, nutrition level response, oxidative stress response, steroid hormone response, transcriptional regulation, transcription factor binding, phosphatase binding, protease binding, and steroid hormone receptor activity were closely related to the common targets. The core targets such as MAPK8, VEGFA and TP53 were related to histone modification; luteolin, quercetin, and kaempferol might be the main drug components affecting histone modification in the treatment of osteoporosis. Results from the in vitro cell experiment revealed that the expression of osteogenesis-related alkaline phosphatase, JMJD2B and RUNX2 protein increased significantly in rat bone marrow mesenchymal stem cells treated with Bushen Jianpi Huoxue Recipe. Histone demethylase JMJD2B, which is closely related to the pharmacological target of Bushen Jianpi Huoxue Recipe, might promote osteogenic differentiation in the treatment of osteoporosis.

Key words: histone demethylase, osteoporosis, JMJD2B, osteogenic differentiation, network pharmacology

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