咳嗽变异性哮喘的差异蛋白膜联蛋白-1的表达及分析  

Analysis of Differential Expression of Annexin-1 Protein in Patients with Cough Variant Asthma

作　　者：邹晓婷[1] 李爽[1] 梁珊珊 王春玲[1] 赵蕴伟 ZOU Xiaoting LI Shuang LIANG Shanshan WANG Chunling ZHAO Yunwei(The First Affiliated Hospital of Jiamusi University, Jiamusi 154002, China)

机构地区：[1]佳木斯大学附属第一医院,黑龙江佳木斯154002

出　　处：《沈阳医学院学报》2017年第5期388-391,共4页Journal of Shenyang Medical College

基　　金：黑龙江省科学技术研究面上项目(No.JMSUJCM2016-041);佳木斯大学研究生科技创新项目(No.YM 2016-018)

摘　　要：目的:研究咳嗽变异性哮喘患者与健康对照者肺泡灌洗液(broncho alveolar lavage fluid,BALF)中有意义的差异蛋白点。方法:选取2016年3月至2016年9月佳木斯大学附属第一医院收治的32例咳嗽变异性哮喘患者为研究对象,作为观察组;选取同期32例健康志愿者为对照组。去除2组研究对象的BALF中的高丰度蛋白,给2组BALF标本分别建立双向凝胶电泳图谱,运用质谱技术得到BALF的蛋白质指纹图谱,筛选出差异表达的蛋白质,再应用生物信息学方法选定其中的一种候选蛋白。结果:凝胶电泳图谱显示,观察组BALF与对照组相比,共有23个蛋白斑点显著异常,经过与质谱分析的指纹图谱进行匹配分析,并结合数据库,初步鉴定出12个蛋白质,其中上调的蛋白质点有:巨噬细胞移动抑制因子(MIF)、程序性细胞死亡基因4(PDCD4)、克拉拉细胞蛋白16(Clara cell protein 16,CC-16)、甲状腺素运载蛋白(transthyretin)、免疫球蛋白结合因子(immunoglobulin binding factor,Ig BF)、半胱氨酸蛋白酶抑制剂S(cystatin S);下调的蛋白质点有:溶菌酶(lysozyme)、蛋白激酶Pak Z、GTP ase Cdc42、膜联蛋白-1(annexin-1)、热休克蛋白Mot-2、乳铁蛋白(lactoferrin)。对下调蛋白膜联蛋白-1进行浓度测定,观察组的膜联蛋白-1浓度显著低于对照组,差异有统计学意义(P<0.05)。结论:用质谱分析鉴定了23个超过3倍差异的蛋白质,其中,共有12个蛋白质被鉴定,生物信息学进一步分析显示膜联蛋白-1能被当成候选蛋白进行更深层次的研究。膜联蛋白-1在哮喘BALF中的表达显著下降,其与哮喘的发病机制密不可分,故能将膜联蛋白-1作为哮喘发病的潜在蛋白标记物。Objective：To investigate the differential expression of proteins in broncho alveolar lavage fluid （BALF） between cough variant asthma patients and healthy controls using two-dimensional gel electrophoresis （2-DE） and mass spectrometry （MS）. Methods：From Mar 2016 to Sep 2016, 32 cases of patients with cough variant asthma were selected as the observation group and 32 healthy volunteers were selected as the control group. BALF were collected from these people and then high-abundance proteins were removed. Then, 2-DE and MS were performed to identify the differentially expressed proteins. Finally, the candidate protein was selected by using bioinformatics method. Results：The results of 2-DE showed that there were 23 differential protein spots between the two groups, among which 12 proteins were identified by MS analysis combined with database. The up-regulated proteins were macrophage migration inhibitory factor （MIF）, programmed cell death 4 gene （PDCD4）, Clara cell protein 16 （CC-16）, transthyretin （TTR）, immunoglobulin binding factor （IgBF）, and cystatin S. And the down-regulated proteins were lysozyme, PakZ, GTPase Cdc42, annexin-1, heat-shock-protein Mot-2, and lactoferrin. The concentration of annexin-1 protein was determined, which was significantly lower in the observation group than that in the control group （p〈0.05）. Conclusions：Twenty-three proteins showing more than 3-fold difference are identified and 12 proteins are furtherly identified by MS. Bioinformatics analysis suggests that annexin-1 could be identified as candidate protein for further study. The expression of annexin-1 decreases significantly in BALF of cough variant asthma patients, which suggests that annexin-1 may be involved in the pathogenesis of asthma, so annexin -1 could be used as a potential marker for asthma.

关 键 词：咳嗽变异性哮喘 双向凝胶电泳技术 质谱技术 生物信息学 膜联蛋白-1 

分 类 号：R562.2[医药卫生—呼吸系统]