Fig 1: Dysregulated BA profiles in the HF-OP group and relative expansion of non-12-OH BAs composition in the HF-OR group. (a) Orthogonal partial least squared-discriminant analysis (OPLS-DA) scores plot of serum BA profiles showing the groupings of N (blue) group, HF-OP (red) and HF-OR (green). (b) VIP scores of OPLS-DA based on the serum BA profiles between the HF-OP and HF-OR group. A BA with VIP more than 1 was considered important in the discrimination between the groups. (c) Dysregulated BAs in the HF-OP group. The data are presented as the mean ± SD. *p<0•05 and **p<0•01 (Mann–Whitney U test). (d) Heatmap of spearman correlation coefficients between serum BAs and blood biochemical parameters from all samples in the three groups (n = 24, 8 samples per group). The gradient colours represent the correlation coefficients, with red color being more positive and blue color indicating more negative. *p<0•05 (Spearman's correlation with the post hoc correction using the Holm method). (e) Differential BAs in liver tissue with a relative expansion of non-12-OH BA composition in HF-OR group. The data are presented as the mean ± SD. *p<0•05 and **p<0•01 (Mann–Whitney U test), ns showed no significance. (f) Pie graphs show the mean percentage of non-12-OH BAs in the liver among the three groups. (g) The relative mRNA levels of Fxr, Shp and critical enzymes responsible for BA synthesis in liver among the three groups. The data are presented as the mean ± SD. **p<0•01 (unpaired Student's t-test). n = 5 per group. (h) The expression of CYP8B1 (n = 4 per group) and CYP7B1 (n = 4 per group) in liver were detected by western blot. Data are presented as the mean ± SD. *p<0•05 and **p<0•01 (unpaired Student's t-test). (i) The expression of FXR and FGF15 in ileum were detected by western blot. Data are presented as the mean ± SD. *p<0•05 (unpaired Student's t-test). (j) Serum FGF15 levels were detected by ELISA. Data are presented as the mean ± SD. **p<0•01 (unpaired Student's t-test).
Fig 2: TH shapes the BA composition by targeting CYP8B1 in the liver.a Heatmaps of all DEGs identified by RNA-seq analysis in the liver of MMI mice treated with T3 for 5 days (left, n = 3) and microarray analysis in the liver of mice upon feeding (right, n = 3). b KEGG analysis of these DEGs in response to T3 treatment and food ingestion, respectively. Venn diagram illustrating commonalities and differences in enriched KEGG pathways regulated by T3 and feeding. c KEGG pathways regulated by T3, which are also regulated by feeding, are ranked by fold enrichment. d, e Relative mRNA levels (d, n = 5) and western blot analysis (e, n = 3) of CYP8B1 in the liver of MMI and MMI + T3-5d mice. f Western blot analysis of CYP8B1 in the liver of Floxed and LTRβKO mice treated with PBS or T3 for 5 days (n = 3). g Western blot analysis of CYP8B1 in the liver of MMI mice and MMI mice treated with MB for 5 days (n = 3). h ChIP-PCR analysis of HA-TRβ recruitment (left) and H3K27 acetylation (right) in the predicted super-enhancer region containing two putative TRβ binding sites (DR1 and DR4) in primary hepatocytes. i Relative mRNA levels of Cyp8b1 in the liver of Floxed and LTRβKO mice treated with T3 (left) or MB (right) for 5 days (n = 5–6). j, k Plasma active GLP-1 (j), plasma insulin and blood glucose levels (k) in MMI and MMI + T3-5d mice administered with AAV-CT or AAV-shCyp8b1 (AAV-sh8B) (n = 5). l oGTT for MMI and MMI + T3-5d mice administered with AAV-CT or AAV-sh8B (left) and the AUC for oGTT (right) (n = 5). m Relative levels of 12α-OH (blue) and non-12α-OH (red) BAs in the gallbladder bile, ileum, liver, serum, and urine of MMI mice and MMI + T3-5d mice (n = 5). n Relative levels of 12α-OH (blue) and non-12α-OH (red) BAs in the ileum of Floxed and LTRβKO mice treated with PBS or T3 for 5 days (n = 5). oGTT oral GTT. Means ± SEM are shown. P values were calculated by two-tailed unpaired Student’s t test. ns not significant. Source data are provided as a Source Data file.
Fig 3: Calcipotriol inhibits ductular reaction and modulates synthesis and transport of BA. (A) Detection of mRNA expression of CK-19 in liver samples with RT-PCR. (B) IHC staining of representative mouse liver samples for CK-19 and image quantification of CK-19 expression. Scale bar: 100 μm. (C) Detection of mRNA expression of CYP7A1, CYP8B1, MRP3, MRP4 and OST-α in liver samples with RT-PCR. (D) Detection of mRNA expression of MRP2, MRP3, MRP4, and OST-α in kidney samples with RT-PCR. All RT-PCR results were normalized against β-actin and expressed as 2-ΔΔCT. All data are presented as mean ± SEM (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).
Fig 4: UDCA treatment attenuated obesity induced by high fat diet. (a) UDCA improved the metabolic profile: changes of body weight and serum paraments. Data are presented as the mean ± SD. n = 5 per group. *p<0•05 and **p<0•01 (unpaired Student's t-test), HFD group vs N group; #p <0•05 and ##p<0•01 (unpaired Student's t-test), HFD+UDCA group vs HFD group. (b) Heatmap of serum bile acids profile of normal diet group (N), high fat diet group (H) and high fat diet + UDCA group (HU). The gradient colours in the heatmap depicted the z-scale value of serum bile acids concentration. (c) The mean percentage of 12-OH bile acids and non-12 bile acids in serum from all the samples of the three group (n = 5 per group). (d) The principal component analysis (PCA) analysis of serum bile acids. n = 5 per group. N: normal diet group; H: high fat diet group; HU: high fat diet group+0•5%UDCA group. (e) Relative mRNA levels of Cyp7a1, Cyp8b1, Cyp27a1 and Cyp7b1 in liver detected by q-PCR assay. All data are presented as the mean ± SD. n = 4 per group. *p<0•05 (unpaired Student's t-test). (f) The expression of CYP8B1 and CYP7B1 in liver of three group mice were detected by western blot. Data are presented as the mean ± SD. n = 3 per group. *p<0•05 (unpaired Student's t-test).
Fig 5: LREE regulates bile acid metabolism through intestinal flora. Red arrows and blue arrows indicate up-regulation and down-regulation after LREE treatment. Normally, most of the primary bile acids are produced by cholesterol through the classical metabolic pathway involving CYP7A1 and CYP8B1. Primary bile acids form conjugated bile acids through the bile duct and are discharged into the intestine. Most of the conjugated bile acids are reabsorbed by the ASBT/OST system and participate in the enterohepatic circulation of bile acids. A small proportion of conjugated bile acids are deconjugated and dehydroxylated by microbiota, converting to secondary bile acids. Different secondary bile acids can activate or inhibit FXR and lead to the release of FGF15. Subsequently, FGF15 entered the liver via the hepatic portal vein to inhibit the expression of CYP7A1 and maintain bile acid levels together with the reabsorbed bile acids. LREE can restore the expression of ASBT, OST-α, FXR, CYP7A1 affected by hyperlipidemia, restore the reabsorption of bile acids, and promote the conversion of cholesterol into bile acids. Meanwhile, LREE increases the synthesis of secondary bile acids and accelerates their excretion through intestinal flora. Increased bile acid metabolism will increase the rate of lipid metabolism and maintain lipid homeostasis in the body.
Supplier Page from Abcam for Anti-CYP8B1 antibody