坑道噪声性听力损失易感者血清差异表达蛋白的筛选及鉴定

Screening and Identification of Differential Proteins of Serum in Individuals Susceptible to Noise Induced Hearing Loss in Tunnel

目的探讨坑道作业部队官兵噪声性听力损失(noise induced hearing loss,NIHL)易感者血清差异表达蛋白的水平及意义。方法选择某部坑道施工部队官兵坑道噪声性听力损失易感组和非易感组各20例作为研究对象,均为男性,易感组平均年龄24.79±2.03岁,高频平均听阈48.55±11.54dB HL,语频平均听阈22.43±8.31dB HL;非易感组平均年龄23.67±3.56岁,高频平均听阈9.40±2.54dB HL,语频平均听阈13.40±4.13dB HL。两组对象分别抽取静脉血5毫升,利用双向电泳(two-dimensional electrophoresis,2-DE)及基质辅助激光解吸电离飞行时间质谱技术(matrix-assisted laser desorption ionization-time of flight mass spectrometry,MALDI-TOF-MS)分类和鉴定两组间血清差异表达蛋白。结果坑道噪声性听力损失易感组和非易感组血清差异表达的蛋白斑点总数为37个,易感组这37个差异蛋白斑点灰度值均值为13 804.27±14 510.62,非易感组为79 820.32±139 333.2,易感组灰度值低于非易感组,差异有统计学意义(t=-2.758,P=0.009 1)。鉴定获得10个差异表达多肽,其中蛋白酶体亚基α-5、补体C4A、载脂蛋白A-I、结合珠蛋白和玻璃体结合蛋白在NIHL易感组表达上调(P<0.05),而甲状腺素运载蛋白、分子量为35kDa胰蛋白酶抑制剂的重链H、色素上皮衍生因子、β-2糖蛋白-I和溶菌酶C表达下调(P<0.05)。结论坑道噪声性听力损失易感者血清中差异表达的蛋白酶体亚基α-5、补体C4A、结合珠蛋白、载脂蛋白A-I、β-2糖蛋白-1、色素上皮衍生因子、胰蛋白酶抑制剂重链H和甲状腺素运载蛋白均与氧化应激反应有关,可能通过该途径参与了NIHL的发生,进而成为坑道作业官兵NIHL易感个体血清特异性候选分子标志物。

Objective To explore the differential expression of protein in the serum of individuals susceptible to noise induced hearing loss(NIHL)susceptible individuals working in the military tunnel.Methods A total of 40 soldiers from one tunnel construction troop were divided into the susceptible group and the nonsusceptible group.Twenty soldiers were selected for each group.The average age of the susceptible group was 24.79±2.03 years old and their thresholds of the speech and high frequencies were 22.43±8.31 dB HL and 48.55±11.54 dB HL,respectively.The average age of the nonsusceptible group was 23.67±3.56 years old and their thresholds of the speech and high frequencies were 13.40±4.13 dB HL and 9.40±2.54 dB HL,respectively.Five microliter peripheral venous blood samples were collected from each individual.Two-dimensional electrophoresis(2-DE)and MALDI-TOF-MS were used to separate and identify the differentially expressed proteins.Results Thirty-seven protein spots differentially expressing between the NIHL susceptible and nonsusceptible were found after 2-DE.Compared by mascort score,10 differential proteins were harvested.Among these,5 peptides including proteasome subunit alpha-5,complement C4-A,haptoglobin,apolipoprotein A-I and vitronectin were upregulated,and other 5 ones,including Lysozyme C,beta-2 glycoprotein-1,pigment epithelium derived factor,35 kDa trypsin inhibitor heavy chain H and transthyretin were downregulated in NIHL susceptible individuals.The differences were statistically significant(P<0.05).Conclusion The differentially expressed proteins were closely related to oxidative stress responses in susceptible individuals,including proteasome subunit alpha-5,complement C4A,haptoglobin,apolipoprotein A-I,beta-2 glycoprotein-1,pigment epithelium derived factor,35 kDa trypsin inhibitor heavy chain H and transthyretin.They might participate in the occurrence of NIHL through this way.The proteins harvested from this study were expected to be specific candidate serum NIHL susceptibility biomarkers in blood to help screen susceptible individuals.