lncRNAs在雄鼠高盐高脂高糖饮食导致的子二代肾功能损害中的作用与机制研究

More and more evidence showed that paternal unhealthy diet and other exposure could affect the occurrence of offspring kidney disease, and lncRNAs may play an important role in them. The applicant's previous study found that male rats with two times high salt-fat-sugar diet exposure induced renal function impairment in the second generation. Eleven candidate lncRNAs were screened by lncRNA microarray analysis and bioinformatics analysis of kidney tissues. However, the role and mechanism of lncRNAs in it need to be further studied. In this study, the 11 lncRNAs will be quantitatively verified, and related mRNAs will be searched, and the serum markers, urine indexes, and renal stereological indices of the second generation will be comprehensively analyzed to determine the target lncRNAs and mRNAs. Molecular biology methods will be used to study the protein-encoding ability, tissue-specificity, subcellular localization and function of the target lncRNAs. The role and molecular mechanism of lncRNAs in the second generation renal function impairment caused by two times exposures of the high salt-fat-sugar diet in male rats were thoroughly investigated. This study can add new materials for the study of paternal programming offspring diseases, provide a new perspective for the prevention and treatment of chronic kidney disease.

越来越多的证据表明父亲不良饮食等暴露因素可以影响子代肾脏疾病的产生，lncRNAs可能在当中发挥重要作用。申请者前期研究发现雄鼠高盐高脂高糖饮食两次暴露导致子二代肾功能损害，经肾脏组织lncRNA芯片检测和生物信息学分析筛选出11个候选lncRNA。然而，lncRNA在其中的作用和机制需深入研究。本研究将对目标lncRNA进行定量验证，寻找相关的mRNA，结合子二代肾功能血清学指标、肾功能尿液指标和肾脏体视学指标进行综合分析，以确定目标lncRNA和mRNA。采用分子生物学手段，对目标lncRNA的蛋白编码能力、组织特异性、亚细胞定位和功能进行研究，深入地探讨lncRNAs在雄鼠高盐高脂高糖饮食两次暴露导致的子二代肾脏功能损害中的作用及分子机制。本研究能为父系编程子代疾病的研究增添新资料，为慢性肾脏疾病的防治提供新视野。

越来越多的证据表明父亲不良饮食等暴露因素可以编程子代肾脏疾病的产生，然而分子机制函待深入研究。因此，本项目开展以下研究工作。研究发现雄鼠交配前高脂高糖高盐饮食两次暴露导致子二代肾功能损害，具有性别差异，以子二代雌鼠肾功能异常更明显。肾小球硬化和肾间质纤维化是肾功能损害重要的病理机制。基因(Enpp6、Tmem144、Actr3b、Cd300lf、 Ifi27、Acadsb)发生差异调控。lncRNA XR-146683.1在子二代雌鼠肾脏中表达量降低，与Tmem144的基因表达呈正相关。但是，lncRNA只在子二代肾功能损害中扮演非常小的角色。DNA甲基化研究显示子二代雌鼠肾脏Cd300lf基因启动子CpG岛的甲基化程度增加，与Cd300lf的基因表达呈负相关。Cd300lf表达与子二代雌鼠的尿白蛋白/肌酐呈负相关。可见由于基因相关的lncRNA和DNA甲基化两方面表观遗传学改变，雄鼠交配前不健康饮食导致子二代易患慢性肾脏疾病。而且，子二代肾功能损害与肠道微生物的属的相对丰度改变相关联。本研究深入地研究了雄鼠交配前高脂高糖高盐饮食暴露编程子代成年期肾功能损害的分子机制，本研究能为父系编程子代疾病的研究增添新资料，为慢性肾脏疾病的防治提供新视野。