O-linked β-N-acetylglucosamine transferase regulates macrophage polarization in diabetic periodontitis: In vivo and in vitro study.

IF 4.2 3区 医学 Q1 ENDOCRINOLOGY & METABOLISM
Ye-Ke Wu, Min Liu, Hong-Ling Zhou, Xiang He, Jing Wei, Wei-Han Hua, Hui-Jing Li, Qiang-Hua Yuan, Yun-Fei Xie
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引用次数: 0

Abstract

Background: Periodontitis, when exacerbated by diabetes, is characterized by increased M1 macrophage polarization and decreased M2 polarization. O-linked β-N-acetylglucosamine (O-GlcNAcylation), catalyzed by O-GlcNAc transferase (OGT), promotes inflammatory responses in diabetic periodontitis (DP). Additionally, p38 mitogen-activated protein kinase regulates macrophage polarization. However, the interplay between OGT, macrophage polarization, and p38 signaling in the progression of DP remains unexplored.

Aim: To investigate the effect of OGT on macrophage polarization in DP and its role in mediating O-GlcNAcylation of p38.

Methods: For in vivo experiments, mice were divided into four groups: Control, DP model, model + short hairpin (sh) RNA-negative control, and model + sh-OGT. Diabetes was induced by streptozotocin, followed by ligation and lipopolysaccharide (LPS) administration to induce periodontitis. The impact of OGT was assessed by injecting sh-OGT lentivirus. Maxillary bone destruction was evaluated using micro-computed tomography analysis and tartrate-resistant acid phosphatase staining, while macrophage polarization was determined through quantitative real-time polymerase chain reaction (qPCR) and immunohistochemistry. For in vitro experiments, RAW264.7 cells were treated with LPS and high glucose (HG) (25 mmol/L D-glucose) to establish a cell model of DP. OGT was inhibited by OGT inhibitor (OSMI4) treatment and knocked down by sh-OGT transfection. M1/M2 polarization was analyzed using qPCR, immunofluorescence, and flow cytometry. Levels of O-GlcNAcylation were measured using immunoprecipitation and western blotting.

Results: Our results demonstrated that M1 macrophage polarization led to maxillary bone loss in DP mice, associated with elevated O-GlcNAcylation and OGT levels. Knockdown of OGT promoted the shift from M1 to M2 macrophage polarization in both mouse periodontal tissues and LPS + HG-induced RAW264.7 cells. Furthermore, LPS + HG enhanced the O-GlcNAcylation of p38 in RAW264.7 cells. OGT interacted with p38 to promote its O-GlcNAcylation at residues A28, T241, and T347, as well as its phosphorylation at residue Y221.

Conclusion: Inhibition of OGT-mediated p38 O-GlcNAcylation deactivates the p38 pathway by suppressing its self-phosphorylation, thereby promoting M1 to M2 macrophage polarization and mitigating DP. These findings suggested that modulating macrophage polarization through regulation of O-GlcNAcylation may represent a novel therapeutic strategy for treating DP.

O-linked β- n -乙酰氨基葡萄糖转移酶调节糖尿病牙周炎巨噬细胞极化:体内和体外研究。
背景:牙周炎,当糖尿病加重时,以M1巨噬细胞极化增加和M2极化减少为特征。由O-GlcNAc转移酶(OGT)催化的o -连接β- n -乙酰氨基葡萄糖(O-GlcNAc酰化)促进糖尿病牙周炎(DP)的炎症反应。此外,p38丝裂原活化蛋白激酶调节巨噬细胞极化。然而,OGT、巨噬细胞极化和p38信号在DP进展中的相互作用仍未被探索。目的:探讨OGT对DP巨噬细胞极化的影响及其介导p38 o - glcn酰化的作用。方法:将小鼠体内实验分为4组:对照组、DP模型、模型+短发夹(sh) rna阴性对照组、模型+ sh- ogt。采用链脲佐菌素诱导糖尿病,结扎和脂多糖(LPS)诱导牙周炎。通过注射sh-OGT慢病毒来评估OGT的影响。通过显微计算机断层扫描分析和抗酒石酸酸性磷酸酶染色评估上颌骨破坏情况,通过定量实时聚合酶链反应(qPCR)和免疫组织化学检测巨噬细胞极化情况。体外实验采用LPS和高糖(HG) (25 mmol/L d -葡萄糖)处理RAW264.7细胞,建立DP细胞模型。OGT抑制剂OSMI4可抑制OGT, sh-OGT转染可抑制OGT。采用qPCR、免疫荧光和流式细胞术分析M1/M2极化。o - glcnac酰化水平采用免疫沉淀和免疫印迹法测定。结果:我们的研究结果表明,M1巨噬细胞极化导致DP小鼠上颌骨丢失,并与o - glcnac酰化和OGT水平升高有关。敲低OGT可促进小鼠牙周组织和LPS + hg诱导的RAW264.7细胞巨噬细胞由M1向M2极化转变。此外,LPS + HG增强了RAW264.7细胞中p38的o - glcn酰化。OGT与p38相互作用,促进其在A28、t141和T347残基上的o - glcn酰化,以及在Y221残基上的磷酸化。结论:抑制ogt介导的p38 o - glcn酰化通过抑制p38自磷酸化使其失活,从而促进巨噬细胞M1向M2极化,减轻DP。这些发现表明,通过调节o - glcn酰化来调节巨噬细胞极化可能是治疗DP的一种新的治疗策略。
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来源期刊
World Journal of Diabetes
World Journal of Diabetes ENDOCRINOLOGY & METABOLISM-
自引率
2.40%
发文量
909
期刊介绍: The WJD is a high-quality, peer reviewed, open-access journal. The primary task of WJD is to rapidly publish high-quality original articles, reviews, editorials, and case reports in the field of diabetes. In order to promote productive academic communication, the peer review process for the WJD is transparent; to this end, all published manuscripts are accompanied by the anonymized reviewers’ comments as well as the authors’ responses. The primary aims of the WJD are to improve diagnostic, therapeutic and preventive modalities and the skills of clinicians and to guide clinical practice in diabetes. Scope: Diabetes Complications, Experimental Diabetes Mellitus, Type 1 Diabetes Mellitus, Type 2 Diabetes Mellitus, Diabetes, Gestational, Diabetic Angiopathies, Diabetic Cardiomyopathies, Diabetic Coma, Diabetic Ketoacidosis, Diabetic Nephropathies, Diabetic Neuropathies, Donohue Syndrome, Fetal Macrosomia, and Prediabetic State.
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