Fig. 1: AGXT is suppressed and LDHA is activated leading to oxalate overproduction in livers from humans and mice with MASH.
a, Schema of glyoxylate metabolism and oxalate formation. Spearman’s correlations were calculated between the expression of genes regulating glyoxylate metabolism or oxalate formation and hepatic fat in livers from transplantation donors (n = 206, GSE26106). A significant inverse correlation is denoted by blue arrows (*P < 0.05, **P < 0.01). The association between the expression of genes regulating glyoxylate metabolism or oxalate formation and MASH was assessed through regression models and a meta-analysis based on transcriptomics of livers from patients with or without MASH (MASH, n = 104; control, n = 44, GSE83452; MASH, n = 24; control, n = 24, GSE61260). A significant inverse association is denoted by purple arrows (**P < 0.01). b, H&E and Picrosirius red staining of liver samples obtained from patients with end-stage MASH (n = 22) compared with healthy donors as control (n = 10). c–g, Expression of AGXT, GRHPR, PRODH2, HOGA1, HAO1 and LDHA relative to GAPDH (c), protein abundance (d) and quantification of AGXT relative to GAPDH (e), from liver samples of MASH patients (n = 23) and control (n = 10), LDH activity, from liver samples of patients with MASH (n = 20) and control (n = 10) (f) and oxalate concentrations normalized to tissue weight in liver samples from patients with end-stage MASH (n = 23) and controls (n = 10) (g). h,i, Liver samples were collected from C57BL/6J mice fed a standard chow diet (control, n = 6) or a high-fat, high-fructose, high-cholesterol diet (MASH diet, n = 6) for 24 weeks (h) and stained with H&E and Picrosirius red (i). j–n, Expression of Agxt, Grhpr, Prodh2, Hoga1, Hao1 and Ldha relative to Gapdh (j), protein abundance (k) and quantification of AGXT relative to β-actin (l), LDH activity (m) and oxalate concentrations normalized to tissue weight in liver samples from mice (n) with (n = 6) or without MASH (n = 6). o, Primary hepatocytes (Hep) from mice fed a standard chow diet and HepG2 cells were treated with either BSA-conjugated PA (200 µM) or BSA control overnight. p, Protein abundance and quantification of AGXT relative to GAPDH (primary hepatocytes, n = 5) or β-actin (HepG2 cells, n = 6). q, Intracellular oxalate normalized to protein concentrations in primary hepatocytes treated with PA (200 µM) or increasing concentrations of sodium oxalate (NaOX; 0–250 µM, n = 4). For primary hepatocytes, each point represents an individual mouse. For HepG2 cells, each point represents an independent experiment that included at least two biological repetitions. The samples derived from the same experiment and blots were processed in parallel for d,e. All data are expressed as mean ± s.e.m. Statistical comparisons were made using two-tailed unpaired t-test (c,f,j,l,m), Mann–Whitney U-test (c,e,g,j,n,p) or one-way ANOVA with Tukey’s multiple comparisons test (q). All individual points and P values are shown. A P value < 0.05 was considered statistically significant; NS, not significant. Scale bars, 200 µm. Parts of h were drawn by using pictures from Servier Medical Art (licensed under a Creative Commons Attribution 3.0 Unported License at https://creativecommons.org/licenses/by/3.0).
