基于LC-MS/MS技术分析中药瓦楞子中发挥免疫调节活性的蛋白质类成分  

Analysis of Immunomodulatory Proteins in Arcae Concha Based on LC-MS/MS Technology

作　　者：石慧 李春磊[1,2] 曾展灏 李欣玥 赵瑞琪 朱建华 于荣敏[1,2] SHI Hui;LI Chun-lei;ZENG Zhan-hao;LI Xin-yue;ZHAO Rui-qi;ZHU Jian-hua;YU Rong-min(College of Pharmacy,Jinan University,Guangzhou 510632,China;Biotechnology Institute of Chinese Materia Medica,Jinan University,Guangzhou 510632,China)

机构地区：[1]暨南大学药学院,广东广州510632 [2]暨南大学中药生物技术研究所,广东广州510632

出　　处：《食品与药品》2022年第3期193-199,共7页Food and Drug

基　　金：国家自然科学基金(编号:82003910);中国博士后科学基金(编号:2020M673092);广东省基础与应用基础研究区域联合基金-青年基金(编号:2020A1515110550);广东省大学生创新创业训练计划(编号:S202010559055)。

摘　　要：目的分析海洋中药瓦楞子中发挥免疫调节作用的蛋白质类成分。方法采用水煎煮和乙酸两种提取方法提取瓦楞子中活性蛋白质类成分;利用RAW264.7巨噬细胞检测瓦楞子提取组分的免疫调节活性;运用瓦楞子转录组测序构建的瓦楞子转录组数据库,通过液相色谱-串联质谱法(LC-MS/MS)分析瓦楞子提取组分的蛋白质类成分;利用GO/KEGG功能富集分析发现瓦楞子提取组分中发挥免疫调节作用的蛋白质类成分。结果得到2个具有免疫调节活性的蛋白质类组分WLZP-1和WLZP-2,其对小鼠RAW264.7细胞均无明显细胞毒性,并能促进RAW264.7细胞分泌一氧化氮(NO),其中,WLZP-1活性优于WLZP-2。同时,分别从WLZP-1和WLZP-2组分中鉴定了276和175个蛋白质类成分。WLZP-1和WLZP-2组分中蛋白质类成分主要由参与分子功能、细胞组分和生物过程的蛋白多肽组成。WLZP-1和WLZP-2组分均具有多个蛋白质类成分参与人源免疫系统及信号传导的功能。结论从瓦楞子两种免疫调节活性组分WLZP-1和WLZP-2中分别发现8个和7个参与人源免疫系统的蛋白质类成分,本研究为发现活性新蛋白多肽及中药瓦楞子免疫调节物质基础研究提供新的科学资料。Objective To analyze the immunoregulatory protein components in Arcae Concha.Methods The active protein components in Arcae Concha were extracted by water decoction and acetic acid extraction.RAW264.7 macrophages were used to detect the immunomodulatory activity of the extracted components of Arcae Concha.Based on the transcriptome database of Arcae Concha constructed by Arcae Concha transcriptome sequencing,the protein compositions of the extracted components of Arcae Concha were analyzed by LC-MS/MS technology.Go/KEGG annotation analysis were used to obtain the material basis of immune regulation in the active proteins of Arcae Concha.Results Two protein components WLZP-1 and WLZP-2 with immunomodulatory activity were obtained.WLZP-1 and WLZP-2 had no obvious cytotoxic effects on RAW264.7 cells,and promoted the production of nitric oxide(NO)in RAW264.7 cells,among which WLZP-1 activity was better than WLZP-2.At the same time,276 and 175 protein compositions were identified from WLZP-1 and WLZP-2,respectively.The protein components of WLZP-1 and WLZP-2 were mainly composed of proteins involved in cell components,molecular functions and biological processes.Both WLZP-1 and WLZP-2 components had multiple protein compositions,which could participate in human immune system and signal transduction.Conclusion Eight and seven proteins involved in human immune system were identified from two protein components WLZP-1 and WLZP-2,respectively.This study provides important scientific data for the discovery of new bioactive proteins and the material basis of immunomodulatory effect from marine traditional Chinese medicine Arcae Concha.

关 键 词：瓦楞子 液相色谱-串联质谱法 免疫调节 蛋白质类成分 

分 类 号：R284.1[医药卫生—中药学]