基于液质联用技术的单胺类神经递质的测定及其在糖尿病中的应用基础研究

Diabetes, as the third biggest killer following cancer and cardiovascular diseases, pose a grave threat to human health. Therefore, carrying out studies about diabetes is very important and urgent. Diabetes is mainly caused by the fact that insulin secreting from pancreatic β-cells is absolutely or relatively deficient. Up to now, the main indicators used for clinical diagnosis and research of diabetes have been insulin, glucose and so on, most of which are downstream effectors. However, substances which might be the potential and predictive markers playing important roles in the upstream are less concerned. As a result, in this study, serotonin and other monoamine neurotransmitters closely related with pancreatic β-cells, which further affects insulin secretion, have been selected as the active markers in upstream for the following research. On the basis of liquid chromatography - mass spectrometry (LC-MS) technique, the methods for sample processing and quantitative analysis have been established. Then, the samples from patients with prediabetes and patients diagnosed with diabetes have been divided into different groups according to ages, and the longitudinal and lateral variation tendency of these monoamine neurotransmitters in different research groups has been studied. These researches lay the theoretical and practical foundation for the establishment of new predictive markers of diabetes. Furthermore, the function of monoamine neurotransmitters in diabetes has been discussed from the qualitative research perspective. This work is expected to provide new ideas and new enlightenment for disease prevention and clinical treatment of diabetes.

糖尿病作为继癌症和心血管疾病之后的第三大杀手，对人类健康有着严重的威胁，因此，开展糖尿病相关方面的研究不仅十分重要，而且迫在眉睫。糖尿病主要是胰岛β-细胞分泌的胰岛素绝对或相对不足所致。目前，用于糖尿病诊断的主要指标是胰岛素和葡萄糖等下游效应物质，而对于其上游作用物质这些潜在的预测标志物关注甚少。本研究中拟选择影响胰岛素分泌，与胰岛β-细胞直接相关的血清素等单胺类神经递质这类上游作用物质作为研究对象，建立基于液相色谱-质谱(LC-MS)技术的样本前处理及定量分析方法，通过研究不同组群前驱和确诊糖尿病患者体内单胺类神经递质的纵向和横向变化规律及这些单胺类神经递质间的联系网络，为糖尿病早期预测标志物的确立奠定理论与实践基础，同时从定量学的角度对单胺类神经递质在糖尿病中的作用进行探讨和阐明，以期为疾病预防和临床治疗提供新思路和新启示。

糖尿病近年来发病率不断上升，根据国际糖尿病联合会最新数据显示：截止2015年，全球大约每11个人中就有一位患有糖尿病，预计到2040年会发展到每10位就有1位糖尿病患者，由此表明，糖尿病及其相关并发症已成为继癌症、心血管疾病之后的人类第三大杀手，严重威胁到人类健康，因此对糖尿病及其并发症的研究显得至关重要。儿茶酚胺是一类单胺类神经递质，作为上游作用物质能影响胰岛素的分泌，且相较其它神经递质更稳定，在糖尿病及其并发症的发生发展中至关重要，但目前其临床检测手段有限，方法灵敏度低、分析速度慢，重现性差，因此建立高效、快速、灵敏和重现性好的方法用于儿茶酚胺的测定十分必要，也为糖尿病及其并发症的研究提供重要手段。. 本研究以儿茶酚胺及其稳定性较好的代谢产物为研究对象，采用液相色谱-串联质谱技术，完成了尿液及血浆中其定量分析方法的建立。此外，我们收集了糖尿病、糖尿病肾病、糖尿病神经病患者及正常对照组的24小时尿液和随机尿液标本，并对其儿茶酚胺进行测定。研究发现儿茶酚胺尽管在糖尿病神经病变患者体内变化并不显著，但在糖尿病转变为糖尿病肾病的过程中扮演着重要角色，其显著降低可能指示糖尿病肾病的存在，由此推测在糖尿病肾病的发病和发展过程中通过减少肾脏交感神经活动有可能达到预防和治疗的效果，为糖尿病并发症的研究提供信息。此外，该研究项目中所建立的基于液质联用技术的儿茶酚胺及其代谢产物测定方法还可用于相关疾病嗜铬细胞瘤的鉴别诊断，已成功推向于我院临床，受到临床一致好评。