基于蛋白质组学技术对竹叶青蛇毒染毒家兔模型血清蛋白质变化的检测及其意义  

Detection of serum protein changes in rabbit model poisoned by Trimeresurus stejnegeri snake venom based on proteomics and its significance

作　　者：杨尾莲[1] 王世军[1] 沈芳华[1] 张勇[1] 陈福伟 苏秋香 史超 余沁瑶 陈涛 YANG Weilian;WANG Shijun;SHEN Fanghua;ZHANG Yong;CHEN Fuwei;SU Qiuxiang;SHI Chao;YU Qinyao;CHEN Tao(Department of Snake Injury,Affiliated People’s Hospital,Fujian University of Traditional Chinese Medicine,Fuzhou 350004,China;Graduate School,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学附属人民医院蛇伤科,福建福州350004 [2]福建中医药大学研究生院,福建福州350122

出　　处：《吉林大学学报（医学版）》2022年第2期308-316,共9页Journal of Jilin University：Medicine Edition

基　　金：国家卫健委中医临床研究基地专项科研项目(JDZX201911);福建省卫健委卫生健康科技计划项目(2020GGA063)。

摘　　要：目的:探讨竹叶青蛇蛇毒模型中血清蛋白的表达,阐明竹叶青蛇蛇毒在染毒过程中标志蛋白的表达情况。方法:将12只雌雄不限日本大耳兔随机分为假手术组和模型组,每组6只。模型组家兔给予20 mg·kg^(-1)竹叶青蛇毒肌肉注射,假手术组家兔给予等量生理盐水注射;4 h后,2组日本大耳兔用戊巴比妥钠麻醉后心脏采血,分离血清,采用绝对定量同位标记(TMT)联合高效液相色谱-串联质谱(HPLC-TMS)定量蛋白组学技术对2组家兔血清进行差异蛋白分析,采用Proteome Discover 2.4等数据库筛选差异蛋白。结果:假手术组与模型组在主成分分析(PCA分析)坐标图上分成明显2簇;差异蛋白鉴定分析共鉴定分析出199个差异蛋白,其中表达上调的蛋白有139个,上调蛋白前5位分别为骨骼肌快肌肌钙蛋白Ⅰ(TNNI2)、小清蛋白α(PV)、烯醇化酶2(ENO2)、肌红蛋白(MB)和肌酸激酶(CK);表达下调蛋白有60个,下调蛋白前5位分别为载脂蛋白C1(APOC1)、扣带蛋白(CGN)、载脂蛋白CI(APOCI)、前蛋白转化酶枯草溶菌素9(PCSK9)和转钴胺素蛋白Ⅰ(TCN1)。基因本体(GO)分析,参与胞外调节功能过程的差异蛋白最多,其次为钙离子调节和蛋白降解调节过程蛋白。结论:竹叶青蛇蛇毒染毒过程中抗原加工及呈递功能有关的蛋白表达水平变化明显,其中CK和溶质载体家族16成员1(SLC16A1)可以作为染毒后损伤的候选靶标。Objective:To investigate the expression of serum proteins of the snake venom model of Trimeresurus stejnegeri,and to reveal the expressions of marker proteins in the process of poisoning.Methods:Twelve Japanese big ear rabbits,male or female,were randomly divided into sham operation group and model group,with 6 rabbits in each group.The rabbits in model group were given intramuscular injection of 20 mg·kg^(-1)Trimeresurus stejnegeri snake venom,and the rabbits in sham operation group were given the same volume of normal saline injection;four hours later,the rabbits in two groups were anesthetized with sodium pentobarbital,the blood samples were obtained from the hearts of rabbits,and the blood serum was isolated.The differential proteins in serum of the rabbits in two groups were analyzed by tandem mass tag(TMT)combined with high performance liquid chromatography-tandem mass spectrometry(HPLC-TMS)quantitative proteomics technology.The biological changes of the differential proteins were analyzed by Proteome Discover 2.4 and other databases.Results:The sham operation group and the experimental group were divided into two distinct clusters of differential proteins on the coordinate diagram of PCA analysis.A total of 199 differential proteins were identified and analyzed,including 139 upregulated proteins;the top 5 up-regulated proteins were troponin I type 2,fast skeletal muscle(TNNI2),parvalbum alpha(PV),enolase2(ENO2),myoglobin(MB)and creatine kinase(CK);there were 60 downregulated proteins;the top 5 down-regulated proteins were apolipoprotein C1(APOC1),cinggulin(CGN),proprotein invertase Bacillus subtilis proteinase 9(PCSK9)and transfer cobalamin protein 1(TCN1).The differential proteins involved in the extracellular regulatory function were the most,followed by calcium ion regulation protein and degradation regulation proteins.Conclusion:The expression levels of antigen processing and presentation function-related proteins change significantly during the process of infection of Trimeresurus stejnegeri venom,an

关 键 词：竹叶青蛇毒 TJMT标记定量蛋白组学 血清差异表达蛋白 肌酸激酶 溶质载体家族16成员1 

分 类 号：R994[医药卫生—毒理学]