基于非标记定量蛋白质组学技术研究重金属铅对水蛭抗凝血活性的影响  

Effect of heavy metal lead on anticoagulant activity of leech based on label-free quantitative proteomics

作　　者：薛淼 孟杰亲 杨超杰 王坤 程亮科 高陆迎 石福娟 麦合丽娅·米吉提 杨瑶珺[1] XUE Miao;MENG Jie-qin;YANG Chao-jie;WANG Kun;CHENG Liang-ke;GAO Lu-ying;SHI Fu-juan;MAIHELIYA Mijiti;YANG Yao-jun(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《中国中药杂志》2021年第22期5804-5809,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81773840)。

摘　　要：该实验以水蛭药材目前在市场上流通最广泛的品种——蚂蟥Whitmania pigra为研究对象,探索重金属铅污染对蚂蟥药材的抗凝血活性的影响,并尝试挖掘其潜在作用机制。该实验利用硝酸铅对蚂蟥养殖土壤进行污染,养殖50 d后得到铅污染组(LC组),同时进行空白对照组(CG组)的养殖。获得蚂蟥样品后,对其进行蛋白提取,通过非标记定量(label-free)蛋白质组学技术分析LC组与CG组的差异表达蛋白。该实验共筛选得到差异表达蛋白152个,其中93个蛋白在LC组中表达上调,59个蛋白表达下调。生物信息学分析发现,差异表达蛋白显著富集在囊泡介导转运和转运正调节等生物过程;在细胞组分方面,显著富集在内吞小泡和顶端质膜等条目;在分子功能方面,显著富集在碳水化合物结合等条目。通过KEGG富集分析,得到差异表达蛋白参与的通路76条,集中体现在代谢途径、次生代谢产物的生物合成和上皮细胞的细菌侵袭等途径。该研究得到2个具有Antistasin结构域的抗凝蛋白,可为进一步探究重金属铅污染对蚂蟥抗凝血活性影响的调控机制和信号转导提供参考。Whitmania pigra is the most widely distributed species of leeches in the market.In this study,the effect of heavy metal lead pollution on the anticoagulant activity of Wh.pigra was studied and the potential mechanism was explored.Pb(NO_(3))_(2) was used to contaminate the breeding soil which was then used to rear Wh.pigra for 50 days(lead-contaminated group,LC group),and meanwhile the blank control group(CG group)was set.Proteins were extracted from the obtained leech samples,and the differentially expressed proteins between LC and CG groups were analyzed by label-free proteomics technology.In this study,a total of 152 differentially expressed proteins were screened out,of which 93 proteins were up-regulated and 59 proteins were down-regulated in LC group.Bioinformatics analysis showed that the biological processes enriched with the differentially expressed proteins were mainly vesicle-mediated transport and transport positive regulation;the enriched cell components were mainly endocytosis vesicles and apical plasma membrane;the enriched molecular functions mainly included carbohydrate binding.The differentially expressed proteins were enriched in 76 KEGG pathways,which mainly involved metabolic pathways,biosynthesis of secondary metabolites,and bacterial invasion of epithelial cells.In this study,two differentially expressed proteins with Antistasin domain were presumed,which provides reference for further exploring the regulatory mechanism and signal transduction underlying the effect of lead pollution on the anticoagulant activity of leech.

关 键 词：蚂蟥 铅污染 蛋白质组学 非标记定量 

分 类 号：R285[医药卫生—中药学]