miRNA质谱分析技术结合多种蛋白质筛选作为OSAHS并发MS相关疾病预警及预后的血清标志物的研究

The incidence of obstructive sleep apnea hypopnea syndrome (OSAHS) is increasing year by year at home and abroad, which has a close relationship with metabolic syndrome(MS)-related diseases, such as hypertension, type 2 diabetes mellitus, hyperlipidemia，and can cause multiple organs and systems damage. Therefore，it will have great significance to look for markers for the early diagnosis and prognosis of OSAHS complicated with MS-related diseases. Our research group has found that serum inflammatory cytokines (CRP, TNF-α, IL-6), oxidative stress-related factor (8-isoprostane), adipokines (Leptin, Adiponectin, Resistin) were elevated in OSAHS patients through combining literature review and our preliminary findings, which were also risk factors for MS -related diseases. In addition，the result of our pilot study has shown that serum miRNAs expressed differences between OSAHS patients and controls. Therefore, this project intends to find serum markers for the early diagnosis and predicting prognosis in OSAHS complicated with MS-related diseases using miRNA mass spectrometry technology combined with a variety of proteins detection. This project also intends to predicate the target genes and their function of differentially expressed miRNAs by bioinformation analysis, which will provide a new perspective to further study the pathogenesis of OSAHS.

阻塞性睡眠呼吸暂停低通气综合征（OSAHS）在国内外人群中发病率逐年升高，其与代谢综合征（MS）相关疾病关系密切，可造成全身多器官多系统损害。因此，寻找可作为OSAHS合并MS相关疾病早期诊断及预后评估的标志物意义重大。本课题组前期研究发现炎症因子、氧化应激相关因子、脂肪因子等MS相关疾病危险因子的血清水平在OSAHS患者中是升高的。另外，预实验结果表明OSAHS患者与正常对照组血清miRNAs存在差异性表达。为此，本项目拟采用miRNA质谱分析技术结合多种蛋白质（1）筛选可作为OSAHS并发MS相关疾病早期诊断的血清标志物。（2）筛选可作为OSAHS并发MS相关疾病预后评估的血清标志物。此外，本项目拟通过生物信息学分析预测差异表达miRNA的靶基因及其功能，为进一步研究OSAHS发病机制提供新的视角。

背景：血清miRNA可以作为代谢相关疾病病程进展及预后的生物标志物，尚未有文献报道miRNA在OSAHS中的变化以及其在OSAHS病程进展中所扮演的角色，前期研究显示OSAHS患者与正常对照者血清存在差异表达miRNA。主要内容：1.采用基因芯片技术获得正常对照者、单纯OSAHS患者、OSAHS合并高血压患者血清miRNA表达谱；2.对比三组的血清miRNA表达谱获得差异性表达的miRNA；3.通过实时荧光定量PCR在更大样本群进行验证；4.运用生物信息学分析对得到验证的血清miRNA进行靶基因及靶基因功能和信号通路的预测；5.运用多元逻辑回归分析、ROC曲线等构建相关疾病预警及诊断模型。6.根据患者的患者资料挖掘睡眠监测指标与临床资料的关系。 重要结果：1.组间比较共筛选出45个差异性表达的miRNA。2.与对照组相比，miR-126-3p在单纯OSAHS组及OSAHS合并高血压组中均表达下降；与对照组相比，let-7d-5p，miR-320b，miR-145-5p在单纯OSAHS组中均表达下降，miR-107, miR-26a-5p在OSAHS合并高血压组中均表达下降。3.组间差异表达miRNA靶基因预测到1307个靶基因，GO分析、Pathway分析显示这些靶基因主要涉及神经系统，代谢，心血管系统，癌症和信号转导等方面。4.重症OSAHS患者，男性，肥胖，白天嗜睡和甘油三酯升高是OSAHS患者出现MSA的危险因素。 科学意义：1.miRNA作为OSAHS相关疾病预警及预后的血清标志物有一定的价值，差异表达的miRNA靶基因及其功能为下一步研究OSAHS发病机制提供新的理论基础。2.完善了血清miRNA在OSAHS中的研究，揭示了OSAHS代谢紊乱病理现象及紊乱点，为OSAHS的治疗提供理论基础。