DNMT3a-mediated methylation of PSTPIP2 enhances inflammation in alcohol-induced liver injury via regulating STAT1 and NF-κB pathway. (March 2022)

• Record Type: Journal Article
• Authors: Xu, Jie-Jie; Zhu, Lin; Li, Hai-Di; Du, Xiao-Sa; Li, Juan-Juan; Yin, Na-Na; Meng, Xiao-Ming; Huang, Cheng; Li, Jun
• Is Part Of: Pharmacological research. Volume 177(2022)
• Publication Date: 2022-03
• Subjects: AAV9 Hepatocyte-Adeno-Associated Virus-9 -- ALD Alcoholic liver diseases -- ALI Alcohol-induced liver injury -- ALT Alanine amino transferase -- AST Aspartate amino transferase -- BSA Bovine serum albumin -- CD Control diet -- ChIP Chromatin immunoprecipitation -- Co-IP Co-Immunocoprecipitation -- CSK C‐terminal Src kinase -- DMEM Dulbecco's modified eagle medium -- DNMTs DNA methyltransferases -- ELISA Enzyme Linked Immunosorbent Assay -- EtOH Ethyl alcohol -- F-BAR Fes/CIP4 homology-Bin/Amphiphysin/Rvs -- FBS Fetal bovine serum -- g gram -- h hour -- H&E Hematoxylin eosin -- IF Immunofluorescence analysis -- IHC Immunohistochemical staining -- IKK IκB kinase -- IL Interleukin -- LPS Lipopolysaccharide -- MSP Methylation-specific PCR -- PB Perfusion buffer -- PSTPIP2 Proline-serine-threonine-phosphatase-interacting protein 2 -- rpm Revolution per minute -- RRBS Reduced Representation Bisulfite Sequencing -- SHIP1 Src homology domain-containing inositol 5′-phosphatase 1 -- STATs Signal transduction and transcriptional activators -- TNF-α Tumor Necrosis Factor-α; Alcohol-induced liver injury (ALI) -- Proline-serine-threonine-phosphatase-interacting protein 2 (PSTPIP2) -- Inflammation -- STAT1 -- DNA methylation; Pharmacology -- Periodicals; Pharmacology -- Periodicals; Research -- Periodicals; Médicaments -- Recherche -- Périodiques; Pharmacologie -- Périodiques; 615.105
• DOI: 10.1016/j.phrs.2022.106125
• Languages: English
• ISSNs: 1043-6618

Abstract

Abstract: Alcohol-induced liver injury (ALI) is associated with inflammatory responses regulated by macrophages. Activation of macrophages plays a crucial role in ALI while DNA methylation-regulated gene silencing is associated with inflammation processes in macrophages. Proline-Serine-Threonine Phosphatase Interacting Protein 2 (PSTPIP2), which belongs to the Fes/CIP4 homology-Bin/Amphiphysin/Rvs domain family of proteins and plays a role in macrophages. Previous studies have shown that Pstpip2 can be methylated. Herein, its expression was found to be significantly downregulated in primary liver macrophages isolated from EtOH-fed mice and EtOH-induced RAW264.7 cells. Overexpression of PSTPIP2 using liver-specific recombinant AAV serotype 9 (rAAV9)-PSTPIP2 in EtOH-fed mice dramatically alleviated liver injury and inflammatory responses. In addition, silencing of PSTPIP2 aggravated the alcohol-induced inflammatory response in vitro . Mechanistically, PSTPIP2 might affect macrophage-induced inflammatory responses by regulating the STAT1 and NF-κB signaling pathways. The downregulation of PSTPIP2 in ALI may be associated with DNA methylation. Methylation-specific PCR and western blotting analyses showed that EtOH induced abnormal DNA methylation patterns and increased the protein expression levels of DNMT1, DNMT3a, and DNMT3b. The chromatin immunoprecipitation assay showed that DNMT3a could directly bind to the Pstpip2 promoter and act as a principal regulator of PSTPIP2 expression. Moreover, silencing of DNMT3a significantly restored the EtOH-induced low expression of PSTPIP2 and inhibited EtOH-induced inflammation. Overall, these findings provide a detailed understanding of the possible functions and mechanisms of PSTPIP2 in ALI, thus providing new substantive research to elucidate the pathogenesis of ALI and investigate potential targeted treatment strategies. Graphical Abstract: ga1