NHE1 regulation in NAFLD in vitro contributes to hepatocyte injury and HSC crosstalk

J Endocrinol. 2024 Oct 8;263(2):e240099. doi: 10.1530/JOE-24-0099. Print 2024 Nov 1.

Abstract

Non-alcoholic fatty liver disease (NAFLD) is the fastest-growing cause of liver-associated death globally. Whole-body knockout (KO) of Na+/H+ exchanger 1 (NHE1, SLC9A1) was previously proposed to protect against high-fat diet-induced liver damage; however, mechanistic insight was lacking. The aim of the present work was to address this question in vitro to determine how NHE1, specifically in hepatocytes, impacts lipid overload-induced inflammation, fibrosis, and hepatocyte-hepatic stellate cell (HSC) crosstalk. We induced palmitate (PA)-based steatosis in AML12 and HepG2 hepatocytes; manipulated NHE1 activity pharmacologically and by CRISPR/Cas9-mediated KO and overexpression; and measured intracellular pH (pHi), steatosis-associated inflammatory and fibrotic mediators, and cell death. PA treatment increased NHE1 mRNA levels but modestly reduced NHE1 protein expression and hepatocyte pHi. NHE1 KO in hepatocytes did not alter lipid droplet accumulation but reduced inflammatory signaling (p38 MAPK activity), lipotoxicity (4-HNE accumulation), and apoptosis (poly-ADP-ribose-polymerase-1 (PARP) cleavage). Conditioned medium from PA-treated hepatocytes increased the expression of NHE1 and of the fibrosis regulator tissue inhibitor of matrix metalloproteinases-2 in LX-2 HSCs, in a manner abolished by NHE1 KO in hepatocytes. We conclude that NHE1 is regulated in NAFLD in vitro and contributes to the ensuing damage by aggravating hepatocyte injury and stimulating hepatocyte-HSC crosstalk.

Keywords: cell biology; fatty acids; intracellular signaling; liver; metabolism.

MeSH terms

  • Animals
  • Hep G2 Cells
  • Hepatic Stellate Cells* / metabolism
  • Hepatocytes* / metabolism
  • Humans
  • Mice
  • Non-alcoholic Fatty Liver Disease* / genetics
  • Non-alcoholic Fatty Liver Disease* / metabolism
  • Sodium-Hydrogen Exchanger 1* / genetics
  • Sodium-Hydrogen Exchanger 1* / metabolism

Substances

  • Sodium-Hydrogen Exchanger 1
  • SLC9A1 protein, human
  • Slc9a1 protein, mouse