Journal: Genes & Diseases

Article Title: Elevated diurnal CD36 expression disrupts the bile acid synthesis rhythm leading to cholestatic liver injury and inflammation via the HMGCR/CYP7A1 axis

doi: 10.1016/j.gendis.2025.101776

Figure Lengend Snippet: Hepatic CD36 was elevated in patients with PBC and PSC, and CD36 displayed abnormally robust diurnal expression in mice with cholestatic liver injury. (A) Schematic representation of the study design for the clinical and animal experiments. The figure was created via BioRender.com. (B) Representative images of liver tissue subjected to immunohistochemistry staining (IHC) for CD36 in normal controls (NCs), PBC patients, and PSC patients. Scale bars: 100 or 500 μm. (C) Western blotting analysis of CD36 expression in the livers of NC ( n = 11), PBC ( n = 5), and PSC ( n = 6) patients. (D) Relative quantification of CD36 protein expression in livers from NC ( n = 11), PBC ( n = 5), and PSC ( n = 6) patients. (E) mRNA expression levels of CD36 in the livers from NC ( n = 11), PBC ( n = 5), and PSC ( n = 6) patients. (F) Linear regression analysis of the correlations between hepatic CD36 mRNA expression and serum ALP, GGT, TBA, and TBIL levels. (G) mRNA expression levels of CD36 in the livers of SHAM and BDL mice ( n = 4 per time point per group) over a 24 h period. (H) Diurnal CD36 protein expression levels from Western blotting analysis of liver tissues from the SHAM and BDL mice ( n = 3 per time point per group). (I) Relative quantification of CD36 protein diurnal expression in liver tissues from the SHAM and BDL mice. (J) Double immunofluorescence staining for CD36 (green) and ALB (red) in the SHAM and BDL mice. Nuclei were counterstained with DAPI (blue). Scale bar: 100 μm. (K) Double immunofluorescence staining for CD36 (green) and CK19 (red) in the SHAM and BDL mice. Nuclei were counterstained with DAPI (blue). Scale bar: 100 μm. (L) Quantitative reverse transcription PCR analysis of CD36 mRNA levels in HepG2 and AML12 cells cultured for 6 h with cholic acid (CA), chenodeoxycholic acid (CDCA), and deoxycholic acid (DCA) at the indicated doses ( n = 6). All the data were presented as mean ± SEM. Group comparisons were performed via two-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group. ZT0 refers to the beginning of the subjective circadian period (6:00 a.m.). The black bars indicate the dark phase from 6:00 p.m. to 6:00 a.m. PBC, primary biliary cholangitis; PSC, primary sclerosing cholangitis; ALP, alkaline phosphatase; GGT, gamma-glutamyl transferase; TBA, total bile acids; TBIL, total bilirubin; BDL, bile duct ligation; ALB, albumin; CD36, cluster of differentiation 36.

Article Snippet: HepG2 (ATCC, USA) and AML12 cells were cultured in Dulbecco's modified Eagle medium (HyClone, USA) supplemented with 10% fetal bovine serum and 100 U/mL penicillin‒streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, Quantitative Proteomics, Double Immunofluorescence Staining, Reverse Transcription, Cell Culture, Control, Ligation