Hypermethylation Effects of Yiqihuoxue Decoction in Diabetic Atherosclerosis Using Genome-Wide DNA Methylation Analyses.
Hypermethylation Effects of Yiqihuoxue Decoction in Diabetic Atherosclerosis Using Genome-Wide DNA Methylation Analyses.
复制标题
利用全基因组 DNA 甲基化分析研究益气活血汤对糖尿病动脉粥样硬化的高甲基化作用
DOI：
10.2147/jir.s335374
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发表时间：
2022
影响因子：
4.5
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Zhou QB;Chen Y;Zhang Y;Li DD;Wang HQ;Jia ZJ;Jin Y;Xu FQ;Zhang Y
目的探讨益气活血方对糖尿病动脉粥样硬化的改善作用及其与DNA甲基化的关系。将载脂蛋白E-敲除小鼠施用链脲佐菌素（50 mg/d，i. p.）高脂饮食喂养16周。将小鼠随机分为DA模型组、罗格列酮组、益气活血低、中、高剂量组。10只健康C57 BL/6 J小鼠为对照组。治疗结束后检测空腹胰岛素、血糖、胰岛素抵抗指数（HOMA-IR）、血脂及炎症因子。采集主动脉组织进行染色（苏木精和伊红以及油红O）。提取基因组DNA用于甲基捕获测序（MC-seq）。使用京都基因和基因组百科全书（KEGG）和检索相互作用基因/蛋白质的搜索工具（STRING）数据库分析差异甲基化基因。焦磷酸测序用于验证MC-seq数据。YQHX低、高剂量组均能降低HOMA-IR（P < 0.05）。与模型组比较，益气活血胶囊低剂量组可降低血清总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、TNF-α、IL-6的表达（P < 0.05）。益气活血汤中剂量组可明显抑制TNF-α的表达（P < 0.05）。YQHX高剂量组可降低IL-6的表达水平（P < 0.05）。免疫组化染色显示益气活血片具有抗动脉粥样硬化作用（P < 0.05）。与对照组相比，MC-seq揭示了DA小鼠中许多异常高甲基化和低甲基化的基因。KEGG数据库分析显示，YQHX处理诱导的高甲基化基因与癌症中的通路、Hippo信号传导和丝裂原活化蛋白激酶相关。网络分析表明，YQHX诱导的表皮生长因子受体（Egfr）和磷酸肌醇-3-激酶调节亚基1（Pik 3r 1）的甲基化在DA中起重要作用;焦磷酸测序结果显示，YQHX显著增加了AKT 1、Nr 1h 3和Fabp 4的甲基化（P <0. 05）。益气活血汤对DA有积极的治疗作用，其机制可能与其调节DNA异常低甲基化有关。
To investigate if a traditional Chinese medicine formulation, called “Yiqihuoxue” (YQHX), could improve diabetic atherosclerosis (DA) and explore potential mechanisms based on DNA methylation. Apolipoprotein E-knockout mice were administered streptozotocin (50 mg/d, i.p.) for 5 days and fed a high-fat diet for 16 weeks. Mice were divided randomly into DA model, rosiglitazone, as well as low-, medium-, and high-dose YQHX groups. Ten healthy C57BL/6J mice were the control group. Serum levels of fasting insulin, blood glucose, homeostasis model-insulin resistance index (HOMA-IR), serum lipids, and inflammatory factors were analyzed after the final treatment. Aorta tissues were collected for staining (hematoxylin and eosin, and Oil red O). Genomic DNA was extracted for methyl-capture sequencing (MC-seq). Kyoto Encyclopedia of Genes and Genomes (KEGG) and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) databases were used to analyze differentially methylated genes. Pyrosequencing was used to verify MC-seq data. Low-dose and high-dose YQHX could reduce the HOMA-IR (P < 0.05). Low-dose YQHX reduced expression of total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), TNF-α, andI L-6 in serum compared with that in the model group (P < 0.05). Medium-dose YQHX decoction inhibited the expression level of TNF-α (P < 0.05). High-dose YQHX decreased the expression level of IL-6 (P < 0.05). Staining also showed the anti-atherosclerosis effects of YQHX (P < 0.05). MC-seq revealed many abnormally hypermethylated and hypomethylated genes in DA mice compared with those in the control group. KEGG database analysis showed that the hypermethylated genes induced by YQHX treatment were related to pathways in cancer, Hippo signaling, and mitogen activated protein kinase. The network analysis suggested that the hypermethylated genes epidermal growth factor receptor(Egfr) and phosphoinositide-3-kinase regulatory subunit 1(Pik3r1) induced by YQHX treatment had important roles in DA. Pyrosequencing revealed that YQHX treatment increased methylation of AKT1, Nr1h3 and Fabp4 significantly (P < 0.05). YQHX decoction had positive treatment effects against DA, because it could regulate aberrant hypomethylation of DNA.