基于lncRNA和环状RNA调控网络的2型糖尿病中医证型表观遗传物质基础研究

Prevention and treatment of diabetes with Traditional Chinese Medicine (TCM) focus on treatment according to symptoms differentiation. However, the distribution of syndromes is complex and lack of objective and quantitative standards. A number of studies have shown that syndromes and type 2 diabetes mellitus (T2DM) are closely related to the epigenetic changes. This project will investigate the correlation between noncoding RNA changes and T2DM in patients of 4 different syndromes (extreme heat due to deficiency of yin, Damp heat trapping spleen, Qi-Yin deficiency and yin-yang deficiency). Using high throughput sequencing technology, we will clarify lncRNA, circRNA and mRNA that are closely related to the occurrence of TCM syndromes of T2DM, providing epigenetic markers for objective quantification of TCM syndrome differentiation in T2DM. We will further identify the key molecules and signaling pathways of differently expressed lncRNAs, circRNAs and their target genes. Finally we will reveal the crosstalk between lncRNA and circRNA in ceRNA networks, and elucidate the epigenetic mechanismin regulating various T2DM syndrome types. The significance of our research is to reveal the epigenetic basis and regulation network for different TCM syndrome types of T2DM , explicit the mechanism of various TCM syndrome types in T2DM and improve the accuracy of clinical dialectical therapy.

中医药防治糖尿病注重辨证论治，但其证候分布规律复杂，目前尚缺乏客观量化的规范标准。多项研究表明，证候和2型糖尿病（T2DM）均与表观遗传关系密切，本课题以阴虚热盛、湿热困脾、气阴两虚、阴阳两虚等4种T2DM中医证型患者为研究对象，采用高通量测序技术： 筛选与T2DM发生及各中医证型密切相关的表观失调的目标lncRNA、circRNA和mRNA，为T2DM中医辨证分型客观量化提供表观遗传标志物；明确差异表达的lncRNAs、circRNAs及其靶基因参与的关键分子和信号通路；揭示lncRNA和circRNA在ceRNA网络中的串扰模式及其调控T2DM各证型基因表达的表观遗传作用机制。意义在于阐明T2DM各中医证型的表观遗传物质基础及其调控网络，为明确T2DM各证型的发生机制，提高临床辨证施治的准确度提供理论依据和数据支持。

中医药防治糖尿病注重辨证论治，但其证候分布规律复杂，目前尚缺乏客观量化的规范标准。多项研究表明，证候和2型糖尿病（T2DM）均与表观遗传关系密切，本课题以阴虚热盛、湿热困脾、气阴两虚、阴阳两虚等4种T2DM中医证型患者为研究对象，采用高通量测序技术：筛选与T2DM发生及各中医证型密切相关的表观失调的目标lncRNA、mRNA以及外显子层面的突变位点，为T2DM中医辨证分型客观量化提供表观遗传及外显子层面标志物。本研究筛选并确定了系列T2DM中医证型特异性分子靶标及其靶基因参与的关键分子和信号通路；我们发现糖尿病患者具有特异性的分子特征，并且差异表达谱与炎症反应和免疫系统密切相关；此外，我们发现T2DM四种中医证型在RNA、DNA和RNA-DNA的表达水平上存在显著差异，湿热困脾型患者在表观遗传修饰和DNA表达水平的分子标志物主要与脂代谢异常相关，气阴两虚型则与线粒体功能降低相关，阴虚热盛型与胰岛素和葡萄糖调节异常相关，阴阳两虚型则与多种糖尿病并发症以及明显的氧化应激密切相关。通过进一步的功能富集分析，我们系统描述了糖尿病不同中医证候的群体特异性生物学特征：湿热困脾型T2DM患者的各种脂质代谢产物水平较高，并且在脂质和氨基酸代谢途径中表现出显著的上调；气阴两虚型患者免疫应答蛋白的许多多肽水平较高；阴阳两虚和阴虚热盛型患者出现PI3K/Akt等代谢途径异常。此外，通过宏基因组学和16SrDNA测序技术研究T2DM4种中医证型的菌群多样性、微生物组成及其功能变化。相关研究成果已发表学术论文5篇，申报专利1项。本研究为明确T2DM各证型的发生机制，提高临床辨证施治的准确度提理论依据和数据支持。