激光显微切割联合质谱分析继发淀粉样变性患者肾组织SAA蛋白  

Analysis of SAA proteins in renal tissue from patients with secondary amyloidosis by laser microdissection and mass spectrometry

作　　者：孙颖[1] 孙健[2] 蔡建芳[1] 文煜冰[1] 郭正光[3] 孙伟[3] 李明喜[1] 李雪梅[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院肾内科,北京100730 [2]中国医学科学院北京协和医学院北京协和医院病理科,北京100730 [3]中国医学科学院北京协和医学院基础医学研究所中心实验室,北京100005

出　　处：《基础医学与临床》2017年第8期1088-1093,共6页Basic and Clinical Medicine

基　　金：国家重点研发计划罕见病临床队列研究(2016YFC0901500)

摘　　要：目的利用激光显微切割联合质谱(LMD/MS)技术分析强直性脊柱炎(AS)合并继发性淀粉样变患者肾活检组织标本血清淀粉样物质A(SAA)蛋白亚型及氨基酸突变序列。方法甲醛固定肾活检组织,脱蜡后行刚果红染色,选取刚果红染色阳性区域进行质谱分析,通过数据分析软件对质谱结果进行整合评估,并将患者SAA蛋白氨基酸序列与变异蛋白数据库氨基酸序列进行比对确定是否有变异蛋白。结果质谱鉴定到高丰度的SAA1及SAA2蛋白,同时有血清淀粉样蛋白P及载脂蛋白E,数据库比对未检测到SAA1及SAA2蛋白的变异序列。结论本研究首次鉴定到了AS合并淀粉样变肾组织中的SAA1及SAA2蛋白,丰富了AS淀粉样变的发病机制,为将来AA型淀粉样变性的精准分型提供新的方法。Objective To analyze serum amyloid protein A （SAA） subtype and amino acid mutation sequence of the renal biopsy specimens from patients with renal amyloidosis secondary to ankylosing spondylitis （AS） by laser microdisseetion combined with mass spectometry. Methods Kidney biopsy formalin-preserved paraffin-embedded （FFPE） specimen slices were stained by Congo red, the positive areas of Congo red staining were selected by mi- crodissection, after trypsin hydrolysis and filtration, peptide samples were subjected to liquid chromatography tan- dem mass spectrometry. Analysis softwares were used to evaluate the results, and the patient＇s amino acid sequence of SAA protein was compared to mutant amino acid sequence reported by literature or deduced from mutant SAA gene to determine whether there was a variation. Results SAA1 and SAA2 proteins with high abundance were identified by mass spectrometry, serum amyloid P and apolipoprotein E were also detected. No variation of SAA1 and SAA2 protein was detected. Conclusions The SAA1 and SAA2 proteins in AA amyloidosis secondary to ASwere identified for the first time, which enriched the pathogenesis of amyloidosis secondary to AS and provided a new method for the accurate classification of AA amyloidosis.

关 键 词：淀粉样变性 激光显微切割 血清淀粉样物质A 液质联用分析法 

分 类 号：R597.2[医药卫生—内科学]