保元汤心肌保护作用靶点群的鉴定与功能分析(1)
[摘要] 保元汤作为治疗元气虚弱的著名方剂，临床上广泛用于冠心病治疗，但由于成分复杂，因此其作用靶点及药理机制尚不明确。该研究构建了一种表面键合有光敏化学基团二苯甲酮的固相微球，利用紫外光激发下二苯甲酮基团与中药化学成分中的碳氢键发生化学偶联反应，将中药保元汤的化学成分组整体键合到微球的表面，并与小鼠心脏组织裂解液共孵育，进而富集保元汤对应的心肌保护靶点群。针对靶点群进行蛋白质组学分析，共发现46个潜在结合靶点蛋白，且大多数定位于线粒体。KEGG分析发现这些靶点蛋白主要与三羧酸循环和氨基酸代谢信号通路关系密切，提示保元汤可能主要通过调控线粒体功能及ATP能量产生进而发挥心肌保护作用。此外JC-1荧光染色实验也证实，保元汤对线粒体损伤具有明显的保护作用。综上所述，该研究通过 “钩钓”直接作用靶点蛋白从源头上解释了保元汤发挥心肌保护作用的潜在药理机制，进而从分子细胞水平上诠释了保元汤的传统功效。此外，该研究也提供了一种用于研究中药复杂方剂多靶点作用机制的新方法，有助于进一步推动传统中药药理机制的现代科学解析。
[关键词] 中药复方； 保元汤； 心肌保护； 靶点； 作用机制
, 百拇医药
[Abstract] Baoyuan decoction (BYD) is a well-known traditional Chinese medicine formula for coronary heart disease with Qi deficiency. However， the detailed pharmacological mechanism of BYD is still unknown because of its complicated chemical compositions. In this study， we synthesized a kind of solid beads with benzophenone groups on its surface. Benzophenone can be activated and chemically cross-linked with the C-H bonds of the chemical compositions in BYD (BYD beads) under UV activation. We thus captured all the target proteins from mouse heart tissue lysates by using BYD beads. Based on proteomics analysis， we discovered totally 46 potential binding target proteins， most of which were located in mitochondria. KEGG analysis revealed that these target proteins were mainly associated with TCA cycle and amino acid metabolism signaling pathways， suggesting that the cardioprotection of BYD might be associated with regulating mitochondrial function and energy production. Moreover， JC-1 staining analysis also confirmed the protective effect of BYD on mitochondrial damage. In summary， our findings elucidated the potential mechanism of BYD on cardioprotection through "target fishing" strategy， and further explained its traditional efficacy in the molecular level. In addition， we also provide an approach for investigating the target group of complicated compositions in Chinese herbal formula. This novel method may provide a methodological reference for exploring the pharmacological mechanism of traditional Chinese formula in the future.
, 百拇医药
[Key words] traditional Chinese formula； Baoyuan decoction； myocardial protection； target； mechanism
中藥复杂方剂在中医药理论指导下，临床疗效确切，但是，由于药效物质基础不明和作用靶点不清，方剂配伍规律的诠释仍停留在经验水平。因此，探究中药复杂体系的作用靶点群，建立一套“基于靶点发现技术”药物机制研究体系，揭示药物作用机制及发现新靶点，对中医药现代化研究具有重要意义[1-2]。
保元汤(Baoyuan decoction，BYD)最早见于明代魏直的《博爱心鉴》[3]，是治疗元气虚弱的著名方剂，主要包含黄芪、人参、甘草(炙)、肉桂4味药材，临床上广泛用于冠心病治疗[4]，经过历代医家数百年的运用验证，保元汤临床疗效确切。由于中药复方是一个复杂体系，因此研究保元汤心肌保护的药理机制就要从多成分、多靶点协同作用的角度进行。本研究利用光化学偶联反应将保元汤的化学成分整体键合到表面修饰有光敏基团二苯甲酮的固相微球上，进而与心脏组织裂解液混合孵育，富集相应的结合蛋白并用高分辨质谱进行鉴定。接着针对每个结合蛋白的功能进行生物信息学分析，阐释保元汤心肌保护作用的分子机制。因此本研究不仅从分子水平上阐释了保元汤多成分、多靶点的药理作用机制，同时对其他中药或复杂方剂体系的作用机制研究也具有重要的借鉴意义。, http://www.100md.com(万彦军 廖理曦 刘瑜琦 姜勇 刘莲英 曾克武 屠鹏飞)
[关键词] 中药复方； 保元汤； 心肌保护； 靶点； 作用机制
, 百拇医药
[Abstract] Baoyuan decoction (BYD) is a well-known traditional Chinese medicine formula for coronary heart disease with Qi deficiency. However， the detailed pharmacological mechanism of BYD is still unknown because of its complicated chemical compositions. In this study， we synthesized a kind of solid beads with benzophenone groups on its surface. Benzophenone can be activated and chemically cross-linked with the C-H bonds of the chemical compositions in BYD (BYD beads) under UV activation. We thus captured all the target proteins from mouse heart tissue lysates by using BYD beads. Based on proteomics analysis， we discovered totally 46 potential binding target proteins， most of which were located in mitochondria. KEGG analysis revealed that these target proteins were mainly associated with TCA cycle and amino acid metabolism signaling pathways， suggesting that the cardioprotection of BYD might be associated with regulating mitochondrial function and energy production. Moreover， JC-1 staining analysis also confirmed the protective effect of BYD on mitochondrial damage. In summary， our findings elucidated the potential mechanism of BYD on cardioprotection through "target fishing" strategy， and further explained its traditional efficacy in the molecular level. In addition， we also provide an approach for investigating the target group of complicated compositions in Chinese herbal formula. This novel method may provide a methodological reference for exploring the pharmacological mechanism of traditional Chinese formula in the future.
, 百拇医药
[Key words] traditional Chinese formula； Baoyuan decoction； myocardial protection； target； mechanism
中藥复杂方剂在中医药理论指导下，临床疗效确切，但是，由于药效物质基础不明和作用靶点不清，方剂配伍规律的诠释仍停留在经验水平。因此，探究中药复杂体系的作用靶点群，建立一套“基于靶点发现技术”药物机制研究体系，揭示药物作用机制及发现新靶点，对中医药现代化研究具有重要意义[1-2]。
保元汤(Baoyuan decoction，BYD)最早见于明代魏直的《博爱心鉴》[3]，是治疗元气虚弱的著名方剂，主要包含黄芪、人参、甘草(炙)、肉桂4味药材，临床上广泛用于冠心病治疗[4]，经过历代医家数百年的运用验证，保元汤临床疗效确切。由于中药复方是一个复杂体系，因此研究保元汤心肌保护的药理机制就要从多成分、多靶点协同作用的角度进行。本研究利用光化学偶联反应将保元汤的化学成分整体键合到表面修饰有光敏基团二苯甲酮的固相微球上，进而与心脏组织裂解液混合孵育，富集相应的结合蛋白并用高分辨质谱进行鉴定。接着针对每个结合蛋白的功能进行生物信息学分析，阐释保元汤心肌保护作用的分子机制。因此本研究不仅从分子水平上阐释了保元汤多成分、多靶点的药理作用机制，同时对其他中药或复杂方剂体系的作用机制研究也具有重要的借鉴意义。, http://www.100md.com(万彦军 廖理曦 刘瑜琦 姜勇 刘莲英 曾克武 屠鹏飞)