基于LC-MS的细胞代谢组学技术分别评价艾迪注射液质量标志物组合物的抗肿瘤作用和心脏毒性保护作用

Cytometabolomics Based on LC-MS Was Used to Evaluate the Antitumor and Cardiotoxic Effects of Aidi Injection Quality Marker Compositions

关键词:  艾迪注射液  质量标志物  抗肿瘤作用  心脏毒性保护作用  UHPLC-Q-Exactive Plus Orbitrap HRMS  细胞代谢组学

Key words:  Aidi Injection  quality markers  anti-tumor effect  cardiotoxic protective effect  UHPLC-Q-Exactive Plus Orbitrap HRMS  cellular metabolomics

摘要:

目的 课题组前期从艾迪注射液(AD)中筛选出6个抗肿瘤的Q-markers和6个心脏毒性保护作用的Q-markers ，探索这12个Q-markers是否代表AD原方中的两种治疗作用。方法 基于前期工作基础，将这两种作用的Q-markers分别按照AD原方相同的浓度配制成不同的抗肿瘤Q-markers组合物和不同的心脏毒性保护作用Q-markers组合物，对比AD原方，在Huh7、HUVEC和H9c2细胞上，采用常规CCK8法分别评价组合物的抗肿瘤作用，以及组合物的心脏毒性保护作用。同时采用UHPLC-Q-Exactive Plus Orbitrap HRMS进行细胞代谢组学研究，分析细胞代谢谱，结合PLS-DA模型计算分析平均距离值(DV)，评价抗肿瘤Q-markers组合物与AD原方的相似性，结合PLS-DA模型计算分析相对平均距离值(RDV)，评价心脏毒性保护作用Q-markers组合物与AD原方的相似性。结果 CCK8和细胞代谢组学实验结果趋势基本一致，显示5个抗肿瘤Q-markers组合物（斑蝥素、人参皂苷Re、人参皂苷Rb1、黄芪皂苷II和人参皂苷Rg2）优于6个抗肿瘤Q-markers组合物和AD原方，5个心脏毒性保护作用Q-markers组合物（刺五加苷E、紫丁香苷、人参皂苷Rd、毛蕊异黄酮葡萄糖苷和壬二酸）优于6个Q-markers组合物和AD原方。结论 斑蝥素、人参皂苷Re、人参皂苷Rb1、黄芪皂苷II和人参皂苷Rg2可作为AD抗肿瘤作用的潜在Q-markers，刺五加苷E、紫丁香苷、人参皂苷Rd、毛蕊异黄酮葡萄糖苷和壬二酸可作为AD心脏毒性保护作用的潜在Q-markers，它们具有同等或优于原方的抗肿瘤和心脏毒性保护作用效果。

Abstract:

OBJECTIVE The research team initially screened six anti-tumor Q-markers and six cardiotoxic protective Q-markers from the Aidi injection (AD) to investigate whether these twelve Q-markers represent the two therapeutic effects found in the original formulation of AD. Methods Based on the preliminary work, the Q-markers with these two functions will be formulated into different anti-tumor Q-marker compositions and various cardiotoxic protective Q-marker compositions, using the same concentration as the original AD formula. In comparison to the original AD formula, the anti-tumor effects of the compositions will be evaluated on Huh7, HUVEC, and H9c2 cells using the conventional CCK8 method, as well as their cardiotoxic protective effects. Additionally, UHPLC-Q-Exactive Plus Orbitrap HRMS will be employed for cellular metabolomics research to analyze the cellular metabolic profiles. The average distance value (DV) will be calculated using the PLS-DA model to assess the similarity between the anti-tumor Q-marker compositions and the original AD formula, while the relative average distance value (RDV) will be calculated to evaluate the similarity of the cardiotoxic protective Q-marker compositions to the original AD formula. Results The results from the CCK8 assay and cellular metabolomics experiments are largely consistent, indicating that the combination of five anti-tumor Q-markers (cantharidin, ginsenoside Re, ginsenoside Rb1, astragaloside II, and ginsenoside Rg2) outperforms the combination of six anti-tumor Q-markers and the original AD formula. Additionally, the five Q-markers with cardioprotective effects (eleutheroside E, syringin, ginsenoside Rd, isoflavone glycoside, and nonanedioic acid) also demonstrate superior efficacy compared to the six Q-markers combination and the original AD formula. Conclusion The compounds cantharidin, ginsenoside Re, ginsenoside Rb1, astragaloside II, and ginsenoside Rg2 may serve as potential Q-markers for the anti-tumor effects in AD. Additionally, eleutheroside E, syringin, ginsenoside Rd, isoflavone glucoside, and nonanedioic acid could act as potential Q-markers for the cardiotoxic protective effects against AD. These compounds exhibit comparable or superior anti-tumor and cardiotoxic protective efficacy compared to the original formulation.