Cytochrome P450 7A1 Antibody - #DF2612

Figure 4 GPA promotes cholesterol metabolism by accelerating the synthesis and efflux of primary bile acids. A) Dynamic distribution map of differences in lipid content. B) Differential lipid KEGG classification map. C) Differential lipid KEGG enrichment map. D) Primary free bile acid content in mouse liver (n = 6). E) Mouse liver primary conjugated bile acid content (n = 6). F) Mouse liver secondary conjugated bile acid content (n = 6). G) Secondary free bile acid content in mouse liver (n = 6). H,I) The mRNA and protein expression of hepatic BSEP, CYP7A1, CYP8B1, CYP27A1, and CYP7B1 in TP chronic DILI mice treated with GPA (n = 6). All mRNA levels were measured by qPCR, normalized to GAPDH. J,K) The mRNA and protein expression of hepatic BSEP, CYP7A1, CYP8B1, CYP27A1, and CYP7B1 in APAP acute DILI mice treated with GPA (n = 6). All mRNA levels were measured by qPCR, normalized to GAPDH. L,M) The mRNA and protein expression of hepatic BSEP, CYP7A1, CYP8B1, CYP27A1, and CYP7B1 in TP chronic DILI mice treated with GPA (n = 6). All mRNA levels were measured by qPCR, normalized to GAPDH. N) Summary of upregulating CYPs-transporter-mediated primary bile acid synthesis and transport of GPA treatment of acute and chronic DILI. All data are presented as means ± SD. Compared with control group, *P < 0.05, **P < 0.01, ***P < 0.001. Compared with model group, #P < 0.05, ##P < 0.01, ###P < 0.001. All data are via one-way analysis of variance (ANOVA).

Fig. 7 Hepatic miR-182-5p targets Cyp7a1 to promote hepatocyte proliferation. a The heatmap of DEGs in BA synthesis signaling pathway. Green represents downregulation while red represents up regulation. qRT-PCR analyses of Cyp7a1 and Cyp27a1 gene expression in the liver of miR-182-5p KO (b) or TG mice (c) compared with their respective control mice (3d after PH; n = 4/group). miR-182-5p mimic (182 m) or its negative control (ncm) were overexpressed in primary hepatocytes from C57BL/6 J mice. d, e miR-182-5p level, and the Cyp7a1 and Cyp27a1 genes expression levels were determined by qRT-PCR (n = 5/group). f The Cyp7a1 protein expression level was determined by western blot. g Alignment of the sequences of Cyp7a1 gene promoter and miR-182-5p. h Luciferase reporter assay to examine the interactions between miR-182-5p and the predicted target site in the Cyp7a1 gene promoter. Plasmids with the Cyp7a1 gene promoter or mutated gene promoter were co-transfected with miR-182-5p mimic or control mimic into primary hepatocytes. Renilla luciferase activity was measured by a Dual-Glo luciferase assay system and normalized to internal control firefly luciferase activity (n = 3 biological Replicates). HSCs co-cultured with Cyp7a1 siRNA (HepCyp7a1-siRNA) or their control siRNA-treated (HepNC-siRNA) hepatocytes isolated from TG mice. i, j qRT-PCR analyses of α-SMA and Ihh genes expression in HSCs. k qRT-PCR analyses of Cyclin genes expression in primary hepatocytes. l EdU immunostaining of primary hepatocytes. m A proposed model on the mechanism by which miR-182-5p regulates liver regeneration induced by PH. Error bars in all experiments represent SEM; Significance was determined by unpaired two-tailed Student’s t test and by one-way ANOVA.