微RNA-34a和启动子甲基化有助于2型糖尿病患者过氧化物酶体增殖激活受体γ基因的表达。

IF 4.3 Q1 ENDOCRINOLOGY & METABOLISM

Diabetes & Metabolic Syndrome-Clinical Research & Reviews Pub Date : 2024-10-01 DOI:10.1016/j.dsx.2024.103156

Mona Moghadasi , Mozhgan Taherimoghaddam , Esmaeel Babaeenezhad , Mehdi Birjandi , Mozhgan Kaviani , Mostafa Moradi Sarabi

{"title":"微RNA-34a和启动子甲基化有助于2型糖尿病患者过氧化物酶体增殖激活受体γ基因的表达。","authors":"Mona Moghadasi , Mozhgan Taherimoghaddam , Esmaeel Babaeenezhad , Mehdi Birjandi , Mozhgan Kaviani , Mostafa Moradi Sarabi","doi":"10.1016/j.dsx.2024.103156","DOIUrl":null,"url":null,"abstract":"<div><h3>Aims</h3><div>This study aimed to investigate the roles of DNA methylation and miR-34a in the regulation of peroxisome proliferator-activated receptor gamma (<em>PPARγ</em>) in patients with type 2 diabetes (T2D).</div></div><div><h3>Methods</h3><div>We investigated the methylation status of four regions of the <em>PPARγ</em> promoter and <em>PPARγ</em> expression in a panel of 84 T2D patients using methylation-specific PCR (MSP) and RT-qPCR, respectively. Moreover, we quantified DNA methyltransferases (<em>DNMTs</em>) expression and global DNA methylation levels by RT-qPCR and ELISA, respectively. We measured the expression levels of miR-34a and protein expression of PPARγ by stem-loop RT-qPCR and ELISA, respectively.</div></div><div><h3>Results</h3><div>We found significant DNA hypermethylation in the R2 and R3 regions of the <em>PPARγ</em> promoter in people with diabetes. Functionally, this was associated with a significant reduction in <em>PPARγ</em> expression. In addition, we observed a significant increase in 5-methylcytosine levels in people with diabetes. A marked increase in circulating miR-34a in the early stages of T2D (up to 10 years) and a significant decrease in circulating miR-34a with increasing diabetes duration from 10 years after the onset of diabetes. Interestingly, upregulation of DNA methyltransferases 1 (<em>DNMT1</em>), <em>DNMT3A</em>, and <em>DNMT3B</em> was observed in people with diabetes, and the average expression of <em>DNMTs</em> was negatively correlated with circulating miR-34a levels. In contrast, the serum protein level of PPARγ, a direct target of miR-34a, increased considerably with diabetes duration and showed a negative correlation with circulating miR-34a, cholesterol, triglyceride, and low-density lipoprotein.</div></div><div><h3>Conclusion</h3><div><em>PPARγ</em> promoter hypermethylation and miR-34a upregulation are associated with T2D pathogenesis through <em>PPARγ</em> dysregulation.</div></div>","PeriodicalId":48252,"journal":{"name":"Diabetes & Metabolic Syndrome-Clinical Research & Reviews","volume":"18 10","pages":"Article 103156"},"PeriodicalIF":4.3000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"MicroRNA-34a and promoter methylation contribute to peroxisome proliferator-activated receptor gamma gene expression in patients with type 2 diabetes\",\"authors\":\"Mona Moghadasi , Mozhgan Taherimoghaddam , Esmaeel Babaeenezhad , Mehdi Birjandi , Mozhgan Kaviani , Mostafa Moradi Sarabi\",\"doi\":\"10.1016/j.dsx.2024.103156\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Aims</h3><div>This study aimed to investigate the roles of DNA methylation and miR-34a in the regulation of peroxisome proliferator-activated receptor gamma (<em>PPARγ</em>) in patients with type 2 diabetes (T2D).</div></div><div><h3>Methods</h3><div>We investigated the methylation status of four regions of the <em>PPARγ</em> promoter and <em>PPARγ</em> expression in a panel of 84 T2D patients using methylation-specific PCR (MSP) and RT-qPCR, respectively. Moreover, we quantified DNA methyltransferases (<em>DNMTs</em>) expression and global DNA methylation levels by RT-qPCR and ELISA, respectively. We measured the expression levels of miR-34a and protein expression of PPARγ by stem-loop RT-qPCR and ELISA, respectively.</div></div><div><h3>Results</h3><div>We found significant DNA hypermethylation in the R2 and R3 regions of the <em>PPARγ</em> promoter in people with diabetes. Functionally, this was associated with a significant reduction in <em>PPARγ</em> expression. In addition, we observed a significant increase in 5-methylcytosine levels in people with diabetes. A marked increase in circulating miR-34a in the early stages of T2D (up to 10 years) and a significant decrease in circulating miR-34a with increasing diabetes duration from 10 years after the onset of diabetes. Interestingly, upregulation of DNA methyltransferases 1 (<em>DNMT1</em>), <em>DNMT3A</em>, and <em>DNMT3B</em> was observed in people with diabetes, and the average expression of <em>DNMTs</em> was negatively correlated with circulating miR-34a levels. In contrast, the serum protein level of PPARγ, a direct target of miR-34a, increased considerably with diabetes duration and showed a negative correlation with circulating miR-34a, cholesterol, triglyceride, and low-density lipoprotein.</div></div><div><h3>Conclusion</h3><div><em>PPARγ</em> promoter hypermethylation and miR-34a upregulation are associated with T2D pathogenesis through <em>PPARγ</em> dysregulation.</div></div>\",\"PeriodicalId\":48252,\"journal\":{\"name\":\"Diabetes & Metabolic Syndrome-Clinical Research & Reviews\",\"volume\":\"18 10\",\"pages\":\"Article 103156\"},\"PeriodicalIF\":4.3000,\"publicationDate\":\"2024-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Diabetes & Metabolic Syndrome-Clinical Research & Reviews\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1871402124002170\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ENDOCRINOLOGY & METABOLISM\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Diabetes & Metabolic Syndrome-Clinical Research & Reviews","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1871402124002170","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ENDOCRINOLOGY & METABOLISM","Score":null,"Total":0}

引用次数: 0

摘要

目的：本研究旨在探讨DNA甲基化和miR-34a在调控2型糖尿病（T2D）患者过氧化物酶体增殖激活受体γ（PPARγ）中的作用：方法：我们使用甲基化特异性 PCR（MSP）和 RT-qPCR 分别研究了 84 例 T2D 患者中 PPARγ 启动子四个区域的甲基化状态和 PPARγ 的表达。此外，我们还通过 RT-qPCR 和 ELISA 分别量化了 DNA 甲基转移酶（DNMTs）的表达和全局 DNA 甲基化水平。我们通过干环 RT-qPCR 和 ELISA 分别测定了 miR-34a 的表达水平和 PPARγ 的蛋白表达：结果：我们发现糖尿病患者 PPARγ 启动子的 R2 和 R3 区域存在明显的 DNA 高甲基化。在功能上，这与 PPARγ 表达的显著降低有关。此外，我们还观察到糖尿病患者体内的 5-甲基胞嘧啶水平明显增加。在 T2D 早期阶段（长达 10 年），循环 miR-34a 明显增加，而从糖尿病发病后 10 年开始，随着糖尿病病程的延长，循环 miR-34a 明显减少。有趣的是，在糖尿病患者中观察到 DNA 甲基转移酶 1（DNMT1）、DNMT3A 和 DNMT3B 的上调，而 DNMTs 的平均表达与循环 miR-34a 水平呈负相关。相反，miR-34a 的直接靶标 PPARγ 的血清蛋白水平随着糖尿病持续时间的延长而显著增加，并与循环 miR-34a、胆固醇、甘油三酯和低密度脂蛋白呈负相关：结论：PPARγ启动子高甲基化和miR-34a上调通过PPARγ失调与T2D发病相关。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

MicroRNA-34a and promoter methylation contribute to peroxisome proliferator-activated receptor gamma gene expression in patients with type 2 diabetes

Aims

This study aimed to investigate the roles of DNA methylation and miR-34a in the regulation of peroxisome proliferator-activated receptor gamma (PPARγ) in patients with type 2 diabetes (T2D).

Methods

We investigated the methylation status of four regions of the PPARγ promoter and PPARγ expression in a panel of 84 T2D patients using methylation-specific PCR (MSP) and RT-qPCR, respectively. Moreover, we quantified DNA methyltransferases (DNMTs) expression and global DNA methylation levels by RT-qPCR and ELISA, respectively. We measured the expression levels of miR-34a and protein expression of PPARγ by stem-loop RT-qPCR and ELISA, respectively.

Results

We found significant DNA hypermethylation in the R2 and R3 regions of the PPARγ promoter in people with diabetes. Functionally, this was associated with a significant reduction in PPARγ expression. In addition, we observed a significant increase in 5-methylcytosine levels in people with diabetes. A marked increase in circulating miR-34a in the early stages of T2D (up to 10 years) and a significant decrease in circulating miR-34a with increasing diabetes duration from 10 years after the onset of diabetes. Interestingly, upregulation of DNA methyltransferases 1 (DNMT1), DNMT3A, and DNMT3B was observed in people with diabetes, and the average expression of DNMTs was negatively correlated with circulating miR-34a levels. In contrast, the serum protein level of PPARγ, a direct target of miR-34a, increased considerably with diabetes duration and showed a negative correlation with circulating miR-34a, cholesterol, triglyceride, and low-density lipoprotein.

Conclusion

PPARγ promoter hypermethylation and miR-34a upregulation are associated with T2D pathogenesis through PPARγ dysregulation.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Diabetes & Metabolic Syndrome-Clinical Research & Reviews ENDOCRINOLOGY & METABOLISM-

CiteScore

22.90

自引率

2.00%

发文量

248

审稿时长

51 days

期刊介绍： Diabetes and Metabolic Syndrome: Clinical Research and Reviews is the official journal of DiabetesIndia. It aims to provide a global platform for healthcare professionals, diabetes educators, and other stakeholders to submit their research on diabetes care. Types of Publications: Diabetes and Metabolic Syndrome: Clinical Research and Reviews publishes peer-reviewed original articles, reviews, short communications, case reports, letters to the Editor, and expert comments. Reviews and mini-reviews are particularly welcomed for areas within endocrinology undergoing rapid changes.