基于队列和代谢组学的职业性噪声暴露生物标志物研究

Based on the occupational noise exposure cohort, this study is to explore the relationship between occupational noise exposure, metabonomics model change and hearing loss by the methods of metabonomics, and further to analyze whether occupational noise exposure can lead to human metabolic abnormalities and these metabolic abnormalities are associated with the development of occupational noise-induced hearing loss. Based on cumulative noise exposure in the noise workers systematically and follow-up, metabolites of the gene coding products associated with noise-induced hearing loss in energy metabolism and oxidative stress pathways are detected and the difference of the metabolites and their concentration in different levels of noise exposure and hearing loss are analyzed . Metabonomics is used to study the metabolic phenotypes of workers exposed to noise and metabolic phenotypes at different levels of hearing loss, thus exploring the metabolic markers of occupational noise-induced hearing loss. The mechanism of noise-induced hearing loss is elucidated by means of further researching the effects of noise exposure and genetic interaction on metabolic markers of occupational noise-induced hearing loss and analyzing the function of susceptible genes and the upstream and downstream relationship of metabolic markers in the metabolic pathway. In addition, the verification of biomarkers and the basic research of transformed application will be carried out, so as to provide theoretical basis and appropriate technology for the three-level prevention, early warning, intervention and early diagnosis of occupational noise-induced hearing loss and better protecting the health of noise-exposed workers.

本研究拟在职业噪声暴露队列人群的基础上，采用代谢组学研究方法，探索职业性噪声暴露、代谢组模式改变和听力损失三者之间的关系，解析职业性噪声暴露导致人体代谢异常，以及这些代谢异常与职业性噪声听力损失发生发展的关系。分析与噪声性听力损失发病风险有关的能量代谢和氧化应激通路中相关基因编码产物在人体内的代谢产物，研究不同噪声暴露水平和不同听力损失水平研究对象代谢组模式特征。应用代谢组学技术研究噪声作业工人的代谢组学表型并分析不同听力损失水平的代谢表型，发现职业性噪声听力损失代谢性标志物。深入研究噪声暴露与基因交互作用对职业性噪声听力损失代谢标志物的影响，通过分析易感基因的功能以及代谢标志物在代谢通路中的上下游关系，阐明噪声导致听力损失的发生机制。开展生物标志物验证以及转化应用的基础研究，为实现职业性噪声听力损失的三级预防、早期预警、干预和早期诊断提供理论依据和适宜技术，保护噪声作业人群健康。

本研究在前期建立的职业噪声暴露队列人群的基础上，采用代谢组学研究方法，探索职业性噪声暴露、代谢组模式改变和听力损失三者之间的关系，解析职业性噪声暴露导致人体代谢异常，以及这些代谢异常与职业性噪声听力损失(NIHL)发生发展的关系。研究随机选取30名噪声暴露且有高频听力损失的工人作为接噪病例组，按照1:1匹配原则从队列中随机选取30名噪声暴露且听力正常的工人作为接噪对照组、30名同工厂中非噪声作业且听力正常的工人作为非接噪对照组，收集三组研究对象的基本信息及血浆样本，用HPLC-MS/MS测定非靶向的血浆代谢组学谱，并对数据进行多变量数据分析、KEGG富集分析和ROC、LASSO分析等。结果显示，鉴定出469种与噪声暴露相关的差异血浆代谢产物及其相关的58种KEGG通路，鉴定出了33种与NIHL相关的差异代谢产物及其相关的12种KEGG途径，其中噪声暴露和NIHL有6种共同的生物学过程。多重比较分析发现，NIHL组存在Pro-Trp、腺嘌呤、二甲基甘氨酸、7-α-羟基脱氢表雄酮、骨化醇、顺式-5-十二烯酸和3β、7-α二羟基-5-胆固醇酸7种代谢产物表达失调。通过LASSO回归分析，构建两个用于NIHL状态预测的风险模型，该模型可以区分NIHL和非NIHL工作者，ROC分析显示，曲线下面积值分别为0.840和0.872，在NIHL诊断中具有较高效率。此外，为验证代谢组学的结果，对易感和耐药小鼠（来自GEO的GSE8342数据集）的耳蜗基因表达进行了比较，结果显示免疫反应和细胞死亡相关过程与噪声暴露关系密切，其在噪声诱导的疾病中发挥着关键作用；根据职业噪声人群累积噪声暴露量，选取24只Wistar雄性大鼠构建噪声暴露大鼠模型，将大鼠与职业人群的差异代谢通路比较取交集发现，醛固酮的合成和分泌、胆汁分泌、癌症中的中心碳代谢、环磷腺苷信号转导途径以及酒精依赖5条代谢通路可能在NIHL及其他噪声相关性疾病中发挥重要作用。研究表明，NIHL病例和非NIHL个体之间的代谢产物谱存在显著差异，噪声暴露可能导致多种生物途径的失调，特别是免疫反应和细胞死亡相关过程。