MicroRNA-320–3p promotes the progression of acute pancreatitis by blocking DNMT3a-mediated MMP8 methylation in a targeted manner. (November 2022)

• Record Type: Journal Article
• Authors: Gu, Huan; Peng, Jie; Wang, Meng; Guo, Zimeng; Huang, Haosu; Yan, Lu
• Is Part Of: Molecular immunology. Volume 151(2022)
• Page Start: 84
• Page End: 94
• Publication Date: 2022-11
• Subjects: Acute pancreatitis -- MicroRNA-320–3p -- DNA methyltransferase 3a -- Matrix metalloprotease 8 -- DNA methylation; Immunochemistry -- Periodicals; Molecular biology -- Periodicals; Immunochemistry -- Periodicals; Allergy and Immunology -- Periodicals; Molecular Biology -- Periodicals; Immunochimie -- Périodiques; Biologie moléculaire -- Périodiques; Immunochemistry; Molecular biology; Periodicals; Electronic journals; 571.96
• DOI: 10.1016/j.molimm.2022.09.003
• Languages: English
• ISSNs: 0161-5890

Abstract

Abstract: In this research, we screened out two genes upregulated in mice with acute pancreatitis (AP) by gene sequencing: microRNA (miR)-320–3p and matrix metalloprotease 8 (MMP8). This study was designed to determine whether miR-320–3p and MMP8 participate in AP development and explore the mechanisms, with a new idea for clinical diagnosis and treatment of AP. Expression of miR-320–3p, DNA methyltransferase 3a (DNMT3a), and MMP8 in mouse pancreatic tissues and AR42J cells was tested by RT-qPCR and western blot assays. Pancreatic pathological changes, serum amylase and lipase, and inflammatory factors in mouse serum and cell supernatant were measured by hematoxylin-eosin staining, automation analyzer, and enzyme-linked immunosorbent assay, respectively. Cell proliferation and apoptosis were determined by CCK-8 assay and flow cytometry. The interaction between miR-320–3p, DNMT3a, and MMP8 was verified by luciferase activity assay, ChIP-qPCR, and MSP assay. High expression of miR-320–3p and MMP8, and low expression of DNMT3a were observed in pancreatic tissues of AP mice and caerulein-induced AP cellular model. Downregulation of miR-320–3p alleviated injury of mouse pancreas, reduced the levels of serum amylase and lipase, and blocked inflammatory factor levels in AP mice. In caerulein-induced AP cellular models, inhibiting miR-320–3p facilitated proliferation and inhibited apoptosis. Upregulation of MMP8 resulted in the opposite results, which could be reversed by simultaneous inhibition of miR-320–3p. miR-320–3p targeted DNMT3a, and downregulating miR-320–3p promoted DNMT3a expression. Moreover, DNMT3a promoted DNA methylation in MMP8 promoter region, thereby inhibiting MMP8 expression in AP mouse and cellular models. This research suggests that miR-320–3p inhibits DNMT3a to reduce MMP8 methylation and increase MMP8 expression, thereby promoting AP progression. Highlights: 1 Upregulated miR-320–3p and MMP8 are found in AP mice and caerulein-induced AP cells. 2 Inhibition of miR-320–3p and MMP8 alleviates the symptoms of AP. 3 Low expression of DNMT3a is found in AP mice and caerulein-induced AP cell model. 4 miR-320–3p targets DNMT3a. 5. DNMT3a promotes MMP8 methylation and reduces MMP8 expression.