基于核磁共振氢谱代谢组学结合分子对接技术筛选补肾壮骨汤促睾酮合成活性成分

引用本文: 吕经纬, 李春楠, 高晓晨, 张楠茜, 张凯月, 赫玉芳, 张辉, 孙佳明. 基于核磁共振氢谱代谢组学结合分子对接技术筛选补肾壮骨汤促睾酮合成活性成分[J]. 分析化学, 2021, 49(5): 779-788. doi: 10.19756/j.issn.0253-3820.201583 shu

Citation: LYU Jing-Wei, LI Chun-Nan, GAO Xiao-Chen, ZHANG Nan-Xi, ZHANG Kai-Yue, HE Yu-Fang, ZHANG Hui, SUN Jia-Ming. Screening of Active Ingredients of Bushen Zhuanggu Decoction for Promoting Testosterone Synthesis Based on ¹H NMR Metabolomics Combined with Molecular Docking Technology[J]. Chinese Journal of Analytical Chemistry, 2021, 49(5): 779-788. doi: 10.19756/j.issn.0253-3820.201583 shu

基于核磁共振氢谱代谢组学结合分子对接技术筛选补肾壮骨汤促睾酮合成活性成分

吕经纬 ^1, ,
李春楠 ^1, ,
高晓晨 ^1, ,
张楠茜 ^1, ,
张凯月 ^1, ,
赫玉芳 ^2, ,
张辉 ^{1,
,} ,
孙佳明 ^{1,
,}

1.
长春中医药大学吉林省人参科学研究院, 长春 130117;
2.
长春中医药大学健康管理学院, 长春 130117

通讯作者: 张辉,E-mail:zhanghui_8080@163.com; 孙佳明,E-mail:sun_jiaming2000@163.com

基金项目:
国家自然科学基金项目（No.82074127）和吉林省科技发展计划项目（No.20200404070YY）资助。

摘要: 基于核磁共振氢谱（¹H NMR）代谢组学结合分子对接技术筛选补肾壮骨汤促睾酮合成的活性成分。应用酶联免疫吸附分析（ELISA）法分析补肾壮骨汤及其膜分离组分对过氧化氢诱导的小鼠睾丸间质细胞（Mouse leydig cells，TM3细胞）上清液睾酮含量的影响，应用超高效液相色谱-四极杆飞行时间质谱（RRLC-Q-TOF-MS）技术快速分析其升睾活性组分中的化学成分组成。利用¹H NMR代谢组学技术揭示过氧化氢诱导TM3细胞在给予升睾活性组分后的差异代谢物，进而通过京都基因与基因组百科全书（KEGG）数据库分析差异代谢物的代谢靶点，并与睾酮合成相关靶点交集，得到升睾活性组分促睾酮合成的效应靶点。最后，确定能够影响效应靶点表达的关键蛋白，应用分子对接技术从活性组分中筛选出与此关键蛋白结合较好的化学成分。在补肾壮骨汤升睾活性组分中解析了27种化学成分的可能结构；细胞代谢组学分析共鉴定出牛磺酸、谷氨酸、乙酸等33种差异代谢物，得到促睾酮合成的效应靶点为睾丸激素17-β-脱氢酶（Testosterone 17-β-dehydrogenase，17β-HSD），KEGG分析表明，抑制关键蛋白α蛋白激酶1（α-kinase 1，ALPK1）磷酸化影响17β-HSD表达能够促进睾酮合成。分子对接结果表明，樱桃苷、原儿茶酸等14个化学成分与ALPK1结合较好，可作为补肾壮骨汤促睾酮合成潜在活性成分，为此复方的药效物质基础研究提供了参考。

关键词:

English

Screening of Active Ingredients of Bushen Zhuanggu Decoction for Promoting Testosterone Synthesis Based on ¹H NMR Metabolomics Combined with Molecular Docking Technology

1.
Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun 130117, China;
2.
School of Health Management, Changchun University of Chinese Medicine, Changchun 130117, China

Corresponding author: ZHANG Hui, ; SUN Jia-Ming,

Received Date: 30 September 2020
Revised Date: 14 January 2021
Fund Project:
Supported by the National Natural Science Foundation of China (No.82074127) and the Jilin Province Science and Technology Development Plan Project (No.20200404070YY).

Abstract: Bushen Zhuanggu Decoctions (BSZGD) is an effective prescription in the treatment of osteoporosis (OP) caused by testosterone deficiency, but the active ingredients for promoting testosterone synthesis are still unclear. The present study aims to reveal the active compounds of BSZGD for promoting testosterone synthesis by using ¹H NMR-based metabolomics and molecular docking methods. After 24 h of administration, BSZGD and its membrane fractions could effectively improve the content of testosterone in the supernatant of Mouse Leydig cells (TM3 cells) induced by hydrogen peroxide. On the basis of medicinal materials, reference materials, literature reports and mass spectrometry data, a total of 27 kinds of chemical composites in the active fractions of BSZGD were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight (RRLC-Q-TOF-MS) technique. ¹H NMR metabolomics technology was used to reveal the differential metabolites of hydrogen peroxide-induced TM3 cells after the active fraction was given. A total of 41 metabolites were identified by ¹H NMR-based metabolomics, of which 33 differential metabolites were found. Testosterone 17-β-dehydrogenase (17β-HSD) as the common metabolic targets of different metabolites and testosterone synthesis was identified using MetScape analysis. KEGG analysis showed that competitive inhibitors of the key protein kinase1 (ALPK1) could activate 17β-HSD and promote the synthesis of testosterone. Fourteen compounds were identified as potential active ingredients in BZGD for promoting testosterone synthesis by molecular docking technique. These results provided further evidence of BSZGD for promoting testosterone synthesis and were beneficial for its clinical application. The potential target and active components found in this study may provide valuable information for further studying the material basis of BSZGD. In addition, this work provided new insights for revealing the active ingredients of complex herbal preparations.

Key words:

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于核磁共振氢谱代谢组学结合分子对接技术筛选补肾壮骨汤促睾酮合成活性成分

通讯作者: 张辉,E-mail:zhanghui_8080@163.com; 孙佳明,E-mail:sun_jiaming2000@163.com

English

Screening of Active Ingredients of Bushen Zhuanggu Decoction for Promoting Testosterone Synthesis Based on ¹H NMR Metabolomics Combined with Molecular Docking Technology

Corresponding author: ZHANG Hui, ; SUN Jia-Ming,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于核磁共振氢谱代谢组学结合分子对接技术筛选补肾壮骨汤促睾酮合成活性成分

通讯作者: 张辉,E-mail:zhanghui_8080@163.com; 孙佳明,E-mail:sun_jiaming2000@163.com

English

Screening of Active Ingredients of Bushen Zhuanggu Decoction for Promoting Testosterone Synthesis Based on 1H NMR Metabolomics Combined with Molecular Docking Technology

Corresponding author: ZHANG Hui, ; SUN Jia-Ming,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

Screening of Active Ingredients of Bushen Zhuanggu Decoction for Promoting Testosterone Synthesis Based on ¹H NMR Metabolomics Combined with Molecular Docking Technology

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