变应性鼻炎冲击免疫治疗血清差异蛋白表达及机制研究

Study on expression and mechanism of serum differential proteins after rush immunotherapy of allergic rhinitis

目的:检测单纯尘螨过敏的变应性鼻炎(AR)患者6 d冲击免疫治疗(RIT)前后相关血清差异蛋白的表达。并对筛选出的CRP、CTHRC1与WDR89三种表达明显差异蛋白进行检测及鉴定,分析并探讨此3种差异蛋白在AR的RIT中的免疫调节作用及意义。方法:收集AR患者、RIT患者及健康对照组血清标本分别为15例、15例、10例,对标本进行等重同位素多标签相对定量蛋白组学技术(iTRAQ)研究,利用二维凝胶电泳及质谱分析确定相关差异蛋白,再利用Cluster3.0软件及Java TreeView软件对所筛选的差异蛋白的合理性进行检验及验证。最后,采用酶联免疫吸附法(ELISA)对筛选出的CRP、CTHRC1与WDR89蛋白进行鉴定。结果:本研究检测到蛋白质总数为893个,发现并确认了53个差异蛋白。AR组与健康对照组相比较,差异有统计学意义(差异倍数>1.2且P<0.05)的差异性表达蛋白有24个,与AR的发生密切相关,其中表达上调的10个,表达下调的14个;RIT组与AR组相比较,差异有统计学意义(差异倍数>1.2且P<0.05)的差异性表达蛋白有29个,与RIT增量阶段的作用相关,其中表达上调的12个,表达下调的17个。与健康对照组相比较,AR组表达上调且进行6 d的RIT,表达下调的蛋白质有CTHRC1、WDR89;与健康对照组相比,AR组表达下调且进行6 d的RIT,表达上调的蛋白质有CRP。采用ELISA对筛选出的CRP、CTHRC1与WDR89蛋白进行鉴定,发现CTHRC1与WDR89差异表达在AR及RIT方面差异有统计学意义(P<0.05),而血清CRP的差异表达在AR及RIT方面差异无统计学意义(P>0.05)。结论:CTHRC1与WDR89和AR发病密切相关,并在RIT中起到一定的调节作用,而血清蛋白CRP在AR及RIT中无明显作用。

Objective:To detect the expression of differentially expressed proteins in serum of patients with allergic rhinitis who were allergic to dust mites before and after 6-day rush immunotherapy.The three differentially expressed proteins,CRP,CTHRC1 and WDR89,were detected and identified.The immunoregulatory effects and significance of these three differentially expressed proteins in rush immunotherapy of allergic rhinitis were analyzed and discussed.Method:The serum samples of 15 patients with allergic rhinitis,15 patients with rush immunotherapy and 10 patients with healthy control group were collected.The samples were studied by isobaric tags for relative and absolute quantitation(iTRAQ)technique.The related differential proteins were determined by two-dimensional gel electrophoresis and mass spectrometry,and the rationality of the screened differential proteins was tested and verified by Cluster3.0 software and Java TreeView software.Finally,the selected CRP,CTHRC1 and WDR89 proteins were identified by enzyme-linked immunosorbent assay(ELISA).Result:In this study,893 proteins were detected and 53 differential proteins were identified.Compared with healthy control group,24 proteins which was statistically significant were found in allergic rhinitis group,which were closely related to the occurrence of allergic rhinitis,including 10 up-regulated proteins and 14 down-regulated proteins.Compared with the allergic rhinitis group,patients with allergic rhinitis underwent 6 days of rush immunotherapy.There were 29 proteins whose expression of proteins with a difference of P value of less than 0.05 and 1.2 times higher,which were related to the effect after the incremental phase of rush immunotherapy was completed,of which 12 were up-regulated and 17 were down-regulated.Compared with healthy control group,the expression of up-regulated of allergic rhinitis group and the expression of down-regulated protein after 6 days of rush immunotherapy were CTHRC1,WDR89;Compared with healthy control group,AR group was down-regulated and the expression of up-regulated protein after 6 days of rush immunotherapy was CRP.CRP,CTHRC1 and WDR89 proteins were identified by enzyme linked immunosorbent assay(ELISA),and it was found that the differential expression of CTHRC1 and WDR89 in AR and RIT was statistically significant(P<0.05),but the differential expression of serum CRP in AR and RIT was not statistically significant(P>0.05).Conclusion:Serum protein CTHRC1 and WDR89 are closely related to the pathogenesis of allergic rhinitis,and played a role in the regulation of rush immunotherapy,while serum protein CRP has no significant effect on AR and RIT.