基于UHPLC-Q Exactive Orbitrap-MS技术的冬虫夏草在不同品系大鼠体内入血成分及代谢产物比较研究  

Comparison of Blood Components and Metabolites of Cordyceps sinensis in Different Strains of Rats Based on UHPLC-Q Exactive Orbitrap-MS Technology

作　　者：刘怡 梁欣欣 乔亚俊 戎林 司鹏龙 魏立新[2,3] 毕宏涛[2,3] Liu Yi;Liang Xinxin;Qiao Yajun;Rong Lin;Si Penglong;Wei Lixin;Bi Hongtao(Department of Pharmacy,Medical College,Qinghai University,Xining 810001,China;Qinghai Provincial Key Laboratory of Tibetan Medicine Pharmacology and Safety Evaluation,Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining 810008,China;Key Laboratory of Tibetan Medicine Research,Chinese Academy of Sciences,Xining 810008,China)

机构地区：[1]青海大学医学院药学系,西宁810001 [2]中国科学院西北高原生物研究所,青海省藏药药理学和安全性评价研究重点实验室,西宁810008 [3]中国科学院藏药研究重点实验室,西宁810008

出　　处：《青海科技》2024年第6期63-76,共14页Qinghai Science and Technology

基　　金：国家自然科学基金面上项目“狭小密闭空间引发精神应激的肠-脑轴时序性调控机制研究”(82171863);青海省创新平台建设专项“青海省糖复合物绿色技术创新服务平台”(2021-ZJ-T02);青海省重大科技专项“冬虫夏草自然资源保育及产业升级绿色关键技术研究”(2021-SF-A4)。

摘　　要：目的 采用UHPLC-Q Exactive Orbitrap-MS技术对冬虫夏草在Lewis大鼠和SD大鼠体内入血成分及代谢产物进行定性分析。方法 采用ACQUITYUPLCHSST3(100 mm×2.1 mm,1.8μm)色谱柱。流动相为0.1%甲酸水溶液(A)和乙腈(B),流速0.35 mL/min,温度45℃。注射量为5μL。进行梯度洗脱。采用加热型电喷雾离子源,正、负离子模式下对冬虫夏草、给药血清和空白血清进行数据采集。此外,将Lewis大鼠和SD大鼠鉴定到的入血成分及代谢产物,通过PharmMapper、Super-Pred平台进行靶点预测和筛选,上传至DAVID平台,进行富集分析。结果 冬虫夏草鉴定出245种化学成分,主要为氨基酸及多肽类、甘油磷脂、脂肪酰类等。Lewis大鼠血清检测到7种成分(入血成分2种、代谢产物5种),SD大鼠血清检测到16种化合物(入血成分2种、代谢产物14种),入血成分主要为生物碱、氨基酸和多肽,代谢产物有多种反应。Lewis大鼠和SD大鼠KEGG和GO富集分析表明,靶点富集在与癌症、免疫相关信号通路,Lewis大鼠中Hepatitis B信号通路与宿主细胞应答相关,SD大鼠中Yersinia infection信号通路影响宿主细胞信号传导。结论 冬虫夏草在Lewis大鼠和SD大鼠中,两者相同的入血成分为gamma-l-glutamyl-l-beta-aminoisibutyricacid,不同的入血成分deltaValerobetaine(Lewis大鼠)、Valylleucine(SD大鼠),两者之间代谢产物的种类及数量均不相同;Lewis大鼠中Hepatitis B信号通路与宿主细胞应答相关,SD大鼠中Yersinia infection信号通路影响宿主细胞信号传导。Aim The blood entry components and metabolites of Cordyceps sinensis in Lewis and SD rats were characterized by UHPLC-Q Exactive Orbitrap-MS technique.Methods The separation was performed on an ACQUITYUPLCHSST3(100mm×2.1mm,1.8µm)column.The mobile phases were 0.1%aqueous formic acid(A)and acetonitrile(B),and the flow rate was 0.35mL/min at 45℃.The injection volume was 5μL.Gradient elution was performed,and the data were collected from Cordyceps sinensis,administered serum and blank serum using a heated electrospray ionization source in positive and negative ion modes.In addition,the incoming components and metabolites identified in Lewis rats and SD rats were subjected to target prediction and screening by Pharm Mapper and Super-Pred platforms,and uploaded to DAVID platform for enrichment analysis.Results By analyzing the mass spectral information of each component,such as the precise relative molecular mass,retention time,secondary fragmentation,and comparing with the mass spectral information of the control product,a total of 245 chemical components were finally identified in Cordyceps sinensis,which were mainly amino acids and peptides,glycerophospholipids,fatty acyls,and other components.In addition,a total of 7 components were detected in Lewis rat serum,including 2 blood-entry components and 5 metabolites,a total of 16 compounds were detected in SD rat serum,including 2 blood-entry components and 14 metabolites.The blood entry components were mainly alkaloids,amino acids and peptides.The metabolites mainly underwent glucuronidation,oxidation,sulfation,hydrolysis,carboxylation and deacetylation,etc.The results of KEGG and GO enrichment analyses in Lewis rats and SD rats indicated that these targets were involved in cancer-and immune-related signaling pathways,such as prostate cancer,PI3K-AKT,and HIF-1,etc.In Lewis rats,the Hepatitis B signaling pathway was associated with host cell response mechanisms;in SD rats,the Yersinia infection signaling pathway affected host cell signaling.Conclusion This method

关 键 词：冬虫夏草 Lewis大鼠 SD大鼠 入血成分 代谢产物 UHPLC-Q Exactive Orbitrap-MS 

分 类 号：R285.5[医药卫生—中药学] R284.1[医药卫生—中医学]