基于iTRAQ技术筛选骨质疏松症患者血清差异蛋白的研究

Research on screening of serum differential proteins in patients with osteoporosis by iTRAQ technology

目的采用同位素相对标记与绝对定量技术(iTRAQ技术)分析骨质疏松症患者和健康人的血清样本中的差异蛋白定量结果,从中筛选并鉴定出骨质疏松症早期诊断的特异性血清蛋白标志物。方法抽取符合入组条件的健康人群和老年性骨质疏松症人群的血清样本各24份,采用iTRAQ技术对各组样本进行蛋白定量定性分析,对比各组样本中蛋白质组的差异性,筛选出存在显著差异(差异倍数>1.2,P<0.01)的特异性蛋白。经查阅uniport数据库及相关文献,分析特异性蛋白的特性及其与骨质疏松症的相关性,以初步鉴定出骨质疏松症早期诊断的特异性血清蛋白标志物。结果通过iTRAQ技术共筛选出31个骨质疏松症相关差异蛋白,其中8个上调,23个下调。筛选出Ephrin A型受体4(EphA4)、角蛋白(KRT)、破骨细胞刺激因子1(OSTF1)作为骨质疏松症患者潜在的生物标志物。结论根据生物通路分析(IPA)结果及各差异蛋白生物生理功能,进一步筛选出EphA4、KRT、OSTF1共3个差异蛋白,初步认定为骨质疏松症早期诊断的潜在蛋白标志物。

Objective To analyze the quantitative results of serum differential proteins in osteoporosis patients and healthy people by isobaric tag for relative absolute quantitation(iTRAQ)technology,and to screen and identify the specific serum protein markers for early diagnosis of osteoporosis.Methods Totally 24 serum samples from healthy people and 24 serum samples from senile patients with osteoporosis were collected.The quantitative and qualitative analysis of protein was carried out in each group by iTRAQ technology,and the differences of proteomics in each group were compared.The specific proteins with significant difference(difference multiple>1.2,P<0.01)were screened.By consulting uniport database and related literatures,the characteristics of specific proteins and their correlations with osteoporosis were analyzed to preliminarily identify specific serum protein markers for early diagnosis of osteoporosis.Results A total of 31 osteoporosis-related differential proteins were screened by iTRAQ technology,of which 8 proteins were up-regulated and 23 proteins were down-regulated.EphA4,keratin(KRT)and osteoclast stimulating factor 1(OSTF1)were selected as potential biomarkers of osteoporosis.Conclusion According to the results of ingenuity pathway analysis(IPA)and the biological and physiological functions of each differential protein,three differential proteins of EphA4,KRT and OSTF1 are further screened and identified as potential protein markers for early diagnosis of osteoporosis.