AGE/RAGE通路microRNA编码基因多态性与2型糖尿病并发冠心病的关联研究

Coronary heart disease (CHD) is the major cause of death in type 2 diabetes (T2DM) complications, advanced glycation end products (AGEs) and receptor of AGEs (RAGE) plays important role in the onset of T2DM induced CHD. We have demonstrated that gene polymorphisms of inflammation pathway were related to cardiovascular disease and its intermediate phenotype, which is in the upstream of AGE/RAGE pathway. Thus, we infer that microRNA related to advanced glycation end products (AGEs) and receptor of AGEs (RAGE) pathway may be associated with T2DM induced CHD. The current study is based on nested case control study design, and chooses T2DM patients as baseline population. The incident CHD patients since 2017 in baseline will be selected as cases, and controls will be chosen matched by sex, age, and duration of diabetes. Questionnaire, physical examination and genotyping is carried out to the study subjects. Genetic risk scores is used to evaluate the association between coding genes of microRNA related to AGEs/RAGE and T2DM induced CHD. Generalized multifactor dimensionality reduction (GMDR) will be used to detect gene-gene and gene-environment interaction. Multiple mediation analysis will be used to infer the possible mechanism of genetic determines. Our study will provide necessary scientific basis for personal secondary prevention of T2DM induced CHD.

冠心病是2型糖尿病（T2DM）并发症的主要死因，晚期糖基化终末产物（AGEs）及其受体（RAGE）通过调控miRNA影响下游通路，是T2DM并发冠心病的始动因素。本课题组前期发现炎性相关基因与心血管疾病相关，据此推测处于炎性通路上游的AGE/RAGE通路相关miRNA编码基因可能与T2DM并发冠心病相关。本研究采用巢式病例对照研究设计，回顾性收集T2DM患者作为研究基线，2017年起对基线研究对象进行随访，将新发冠心病患者纳入病例组，按年龄、性别、T2DM病程、匹配同期对照；检测AGE、RAGE等的血清学水平，并确定易感基因型，综合应用遗传风险评分及多因子降维法评价AGE/RAGE通路microRNA编码基因多态性与T2DM并发冠心病的关联，并通过多元多重中介效应分析推测遗传因素的致病机制，探讨在miRNA层面选择的基因对疾病作用的新模式，可为T2DM并发冠心病的个体化二级预防提供依据。

冠心病是2型糖尿病主要的大血管并发症，本课题通过巢式病例对照研究，探讨晚期糖基化终产物（AGEs）调控通路的miRNA相关基因多态性（SNPs）与糖尿病并发冠心病的关联。研究结果显示，单纯糖尿病患者AGEs水平增高（P=0.021），糖尿病并发冠心病患者IL-6水平增高（P=0.024）。遗传风险评分与疾病关联具有统计学意义，PRS每增加1分，疾病风险增加9.1%（P=0.015）。.综合miRNA测序，候选基因关联研究，多因子降维法（GMDR）及中介效应分析，本课题发现两条可能的糖尿病并发冠心病致病通路：（1）血压、血小板水平为糖尿病患者动脉粥样硬化的危险因素，舒张压（DBP）的间接效应强度最大（IE=3.22），中介效应比例最高（13%）。miR-125a-5p在疾病组上调，其下游rs11881781位点的C等位基因可增加31%的疾病风险（P=0.027），中介效应分析显示，该位点可能通过DBP影响疾病的发生（P=0.042），中介效应占比例35.2%。该结果提示了miRNA-SNP的调控机制，并提示了遗传可能通过影响血压导致疾病，为疾病的早期预防提供了理论依据。（2）miRNA测序结果提示，miR-204-5p、miR-125a-5p、miR-10a-5p等在疾病组异常表达。通过查阅mirTarBase数据库及相关文献，miR-204-5p可靶向作用于IL6R基因，miR-125a-5p可靶向作用于STAT3基因，提示了IL6/STAT3通路对糖尿病并发冠心病的作用。miR-10a-5p靶基因RAGE上的rs184003位点每个A等位基因可增高疾病风险49.1%（P=0.005），miR-204-5p靶基因IL6R上的rs4845625位点同样与疾病具有统计学关联（P=0.003），GMDR法发现RAGE基因rs184003位点与IL6R基因rs4845625位点的交互作用具有统计学意义（CV=10/10，P=0.001），该结果进一步提示，RAGE可能通过激活IL6相关通路进而影响糖尿病并发冠心病的发病。