Journal: Journal of Lipid Research

Article Title: FXR activation by obeticholic acid or nonsteroidal agonists induces a human-like lipoprotein cholesterol change in mice with humanized chimeric liver [S]

doi: 10.1194/jlr.M081935

Figure Lengend Snippet: FXR-mediated increases in LDL-C are also elicited by nonsteroidal FXR agonists in humanized PXB mice. A: Chemical structure of OCA and the two nonsteroidal compounds, INT-2228 and INT-2231, used in this study. B: qPCR analysis of human FXR-associated genes in PXB mouse livers. Significance was determined by one-way ANOVA with Dunnett’s correction (vs. vehicle). n = 5 mice per group. C: Serum cholesterol measurements from PXB mice were determined by gel permeation HPLC. Shown are the cholesterol levels that changed from baseline in response to vehicle, OCA (10 mg/kg/day), and/or atorvastatin (ATV; 10 mg/kg/day) treatment. Significance was determined by one-way ANOVA with Dunnett’s correction (vs. vehicle). n = 5 or 6 mice per group. D: Liver human LDLR gene expression levels of PXB mice measured by qPCR. Significance was determined by one-way ANOVA with Dunnett’s correction (vs. vehicle). n = 5 mice per group. E: Western blot analysis of liver samples from PXB mice treated with vehicle, OCA, INT-2228, and INT-2231. Five biological replicates are shown for each group. F: Protein quantification of Western blots from E. Levels of indicated proteins were normalized to levels of βACTIN. Significance was determined by Student’s t-test. n = 5 mice per group. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Total liver cholesterol and triglycerides (Sekisui Medical Co., Tokyo, Japan) as well as serum enzymes (Fujifilm Corp., Tokyo, Japan) were measured by Skylight Biotech using commercial kits.

Techniques: Gene Expression, Western Blot

Journal: Journal of Lipid Research

Article Title: FXR activation by obeticholic acid or nonsteroidal agonists induces a human-like lipoprotein cholesterol change in mice with humanized chimeric liver [S]

doi: 10.1194/jlr.M081935

Figure Lengend Snippet: PXB mice exhibit a human-like lipoprotein cholesterol profile. A, B: Representative serum lipoprotein measurements by gel permeation HPLC from control (SCID) and PXB mice. The x axes are fractions where the indicated lipoproteins elute. CM, chylomicron. Data points represent mean ± SD of n = 9 mice per group. These mice have been treated with vehicle and are included in the analyses for Figs. 1–4. C: Serum LDL-C/HDL-C ratio calculated by dividing total LDL-C (fractions 8–13) with total HDL-C (fractions 14–20). Significance was determined by Student’s t-test. n = 9 mice in each group. D: Serum VLDL-C/total cholesterol ratio calculated by dividing total VLDL-C (fractions 3–7) with total cholesterol (fractions 1–20). Significance was determined by Student’s t-test. n = 9 mice per group. E: Liver mouse Ffg15 and human FGF19 gene expression analysis by qPCR of control (SCID) and PXB mice, respectively. Control mice: n = 5 mice per group. PXB mice: Significance was determined by Student’s t-test, n = 8 or 9 mice per group. * P < 0.05; **** P < 0.0001.

Article Snippet: Total liver cholesterol and triglycerides (Sekisui Medical Co., Tokyo, Japan) as well as serum enzymes (Fujifilm Corp., Tokyo, Japan) were measured by Skylight Biotech using commercial kits.

Techniques: Control, Gene Expression

Journal: Journal of Lipid Research

Article Title: FXR activation by obeticholic acid or nonsteroidal agonists induces a human-like lipoprotein cholesterol change in mice with humanized chimeric liver [S]

doi: 10.1194/jlr.M081935

Figure Lengend Snippet: Liver cholesterol levels are increased in OCA-treated chimeric mice. A: Measurement of liver cholesterol levels in control (SCID) and PXB mice treated with OCA (10 mg/kg/day) and/or atorvastatin (ATV; 10 mg/kg/day). Total cholesterol was determined by an in vitro enzymatic assay. Control: Changes were not significant by Student’s t-test. n = 9 mice per group. PXB: Significance was determined by one-way ANOVA with Tukey’s correction. n = 8 or 9 mice per group. B: Measurement of liver triglycerides by in vitro enzymatic assay. Changes were not significant by Student’s t-test (Control) or one-way ANOVA (PXB). C: qPCR analysis of human lipogenic gene expression in PXB mouse livers. *** P < 0.001; **** P < 0.0001; n.s., not significant.

Article Snippet: Total liver cholesterol and triglycerides (Sekisui Medical Co., Tokyo, Japan) as well as serum enzymes (Fujifilm Corp., Tokyo, Japan) were measured by Skylight Biotech using commercial kits.

Techniques: Control, In Vitro, Enzymatic Assay, Gene Expression

Journal: Journal of Lipid Research

Article Title: FXR activation by obeticholic acid or nonsteroidal agonists induces a human-like lipoprotein cholesterol change in mice with humanized chimeric liver [S]

doi: 10.1194/jlr.M081935

Figure Lengend Snippet: Cholesterogenic gene expression is reduced by OCA and is normalized by atorvastatin coadministration in humanized mice. A: qPCR analysis of mouse and human FXR-associated genes [CYP7A1, NR0B2 (SHP), SCARB1 (SR-B1), and SLC51B (OSTβ)] in control (SCID) and PXB mouse livers, respectively. Significance was determined by Student’s t-test. Control: n = 5 mice per group; PXB: n = 8 or 9 mice per group. B: qPCR analysis of mouse cholesterol synthesis-associated genes in control (SCID) livers. Significance was determined by Student’s t-test. n = 5 mice per group. C: qPCR analysis of human cholesterol synthesis-associated genes in PXB livers. Significance was determined by one-way ANOVA with Dunnett’s correction (vs. vehicle-treated group). n = 8 or 9 mice per group. D: Correlation between human LDLR mRNA levels and human CYP7A1 mRNA levels in individual PXB mice. Linear regression shown with goodness of fit (R2) value. P value indicates that the slope is significantly nonzero. E: Western blot analysis of liver samples from PXB mice treated with vehicle, OCA (10 mg/kg/day), and/or atorvastatin (ATV; 10 mg/kg/day). Full-length precursor SREBP2 is designated pSREBP2, and its active, mature form is designated mSREBP2. F: Protein quantification of all biological replicates from Western blots from E and supplemental Fig. 3B. Signal is normalized to βACTIN. Significance was determined by one-way ANOVA with Sidak’s correction (vs. OCA-treated group). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., not significant.

Article Snippet: Total liver cholesterol and triglycerides (Sekisui Medical Co., Tokyo, Japan) as well as serum enzymes (Fujifilm Corp., Tokyo, Japan) were measured by Skylight Biotech using commercial kits.

Techniques: Gene Expression, Control, Western Blot

Journal: Journal of Lipid Research

Article Title: FXR activation by obeticholic acid or nonsteroidal agonists induces a human-like lipoprotein cholesterol change in mice with humanized chimeric liver [S]

doi: 10.1194/jlr.M081935

Figure Lengend Snippet: OCA treatment in PXB mice increases serum LDL cholesterol that is attenuated by atorvastatin coadministration. A: Serum lipoprotein measurements from control (SCID) and PXB mice were determined by gel permeation HPLC. Shown are the cholesterol levels that changed from baseline in response to vehicle, OCA (10 mg/kg/day), and/or atorvastatin (ATV; 10 mg/kg/day) treatment. Control: Significance was determined by Student’s t-test. n = 9 mice per group. PXB: Significance was determined by one-way ANOVA with Tukey’s correction. n = 8 or 9 mice per group. B, C: Protein ELISA measurements (absolute values) of indicated lipoproteins from PXB mice. Significance was determined by one-way ANOVA with Tukey’s correction. n = 8 or 9 mice per group. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., not significant.

Article Snippet: Total liver cholesterol and triglycerides (Sekisui Medical Co., Tokyo, Japan) as well as serum enzymes (Fujifilm Corp., Tokyo, Japan) were measured by Skylight Biotech using commercial kits.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: A flavonoid-rich fraction of Euphorbia peplus attenuates hyperglycemia, insulin resistance, and oxidative stress in a type 2 diabetes rat model

doi: 10.3389/fphar.2023.1204641

Figure Lengend Snippet: EPE decreased liver TG (A) and cholesterol (B) , prevented lipid deposition (C) , and ameliorated serum ALT (D) and AST (E) in diabetic rats. Data are mean ± SD ( n = 6). * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. control. ### p < 0.001 vs. diabetic.

Article Snippet: Liver triglycerides (TGs) and cholesterol were assayed using Spinreact (Spain) kits after extracting the lipids using chloroform/MeOH mixture (2:1, v/v) as descried by Folch et al (1957) .

Techniques: Control

Journal: Foods

Article Title: Structural Characterization of Polysaccharide from Flammulina velutipes and Its Impact on Hyperlipidemia Through Modulation of Hepatic Cholesterol Metabolism and Gut Microbiota

doi: 10.3390/foods14193452

Figure Lengend Snippet: Effect of FVPB1 on the expression of cholesterol transport-related factors in mice induced by HFD at the mRNA and enzymes level. ( A – D ) Ldlr mRNA, Hmgcr mRNA, Lcat mRNA, and Cyp7a1 mRNA; ( E – G ) HMG-CoA reductase, LCAT enzyme, and CYP7A1 enzyme. Values are expressed as mean ± SD ( n = 10). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. # indicates a significant difference between ND and HFD ( p < 0.05), while ## indicates an extremely significant difference between ND and HFD ( p < 0.01); * denotes a significant difference compared with HFD ( p < 0.05), while ** represents an extremely significant difference compared with HFD ( p < 0.01). Six treatment groups were as follows: ND, treated with a natural diet group; HFD, treated with a high-fat diet group; SV, HFD treated with 2.5 mg/kg/d simvastatin; HDFL, HFD treated with 100 μg/mL FVPB1; HDFM, HFD treated with 200 μg/mL FVPB1; HDFH, HFD treated with 400 μg/mL FVPB1.

Article Snippet: The levels of TC (Cat No. A111-1-1), TG (Cat No. A110-1-1), LDL-C (Cat No. A113-1-1), HDL-C (Cat No.A112-1-1), serum apolipoprotein A-I (Cat No. E020-1-1), apolipoprotein B (Cat No. E021-1), liver HMGCR (Cat No. H236), liver human CYP7A1 (Cat No. H461-1-2), liver lecithin cholesterol acyltransferase (LCAT, Cat No. H237), and fecal coprostanol (Cat No. A111-1-1) and bile acids (Cat No. 003-2-1) were measured using assay kits (all Jiancheng Biological Engineering Institute, Nanjing, China) following the manufacturer’s instructions.

Techniques: Expressing