fxr agonist 3  (MedChemExpress)

Journal: Gut Microbes

Article Title: Antibiotic cocktail-induced changes in gut microbiota drive alteration of bile acid metabolism to restrain Th17 differentiation through the FXR–NLRP3 axis

doi: 10.1080/19490976.2025.2582944

Figure Lengend Snippet: DCA inhibits FXR activation of the NLRP3-IL17A signaling pathway to promote Th17 cell differentiation (A) Feeding schedule for EAP mice treated with ABX + DCA, ABX + DCA + GW4064, ABX + DCA + MCC950, or ABX + DCA + GW4064 + MCC950. (B) Assessment of pelvic pain using von Frey filaments. (C) H&E staining revealing alterations in prostate tissue. The red arrow indicates the invasion of inflammatory cells. Inflammation score assessment in ABX + DCA, ABX + DCA + GW4064, ABX + DCA + MCC950 and ABX + DCA + GW4064 + MCC950 mice. (D, E) Relative levels of RORγt and FXR mRNA in prostate from the four groups. (F, G) The percentages of CD4+ IL-17A + and CD4+ RORγt + cells in the spleens of ABX + DCA, ABX + DCA + GW4064, ABX + DCA + MCC950 and ABX + DCA + GW4064 + MCC950 mice. (H) Serum concentrations of IL-17A, GM-CSF, and IFN- γ in the four groups detected by ELISA.Sorted naïve CD4+ T cells were activated for 5 days under Th17 cell differentiation conditions without AAV-FXR (media group), with AAV-FXR (media + AAV-FXR group), with AAV-FXR + GW4064 (media + AAV-FXR + GW4064 group), or with AAV-FXR + MCC950 (media + AAV-FXR + MCC950 group). (I) Medium concentrations of IL-17A, GM-CSF, and IFN- γ in the media, media + AAV-FXR, media + AAV-FXR + GW4064 and media + AAV-FXR + MCC950 groups detected by ELISA. (J, K) Flow cytometric analysis of the proportions of CD4+ IL−17 + and CD4+ RORγt + cells in the four groups. (L) Relative mRNA expression of IL-17A, NLRP3, FXR and ASC in the four groups determined via RT‒qPCR. (M) Western blot analysis of NLRP3-IL17A-related proteins and FXR in the media, media + AAV-FXR, media + AAV-FXR + GW4064 and media + AAV-FXR + MCC950 groups. ( N ) IHC of NLRP3-IL17A-related proteins and FXR in prostate tissue from ABX + DCA, ABX + DCA + GW4064, ABX + DCA + MCC950 and ABX + DCA + GW4064 + MCC950 mice. (O) IHC of NLRP3-IL17A-related proteins and FXR in prostate tissue from EAP and EAP + ABX mice. ( P ) IHC of NLRP3-IL17A-related proteins and FXR in the prostate tissue of recipient EAP mice treated with FMT from EAP or EAP + ABX mice ( n = 3–5; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).

Article Snippet: The mice were randomly divided into 5 groups: the antibiotic mixture treatment group (ABX), the antibiotic treatment with diet group (ABX + DCA, 0.2% DCA, MCE, HY-N0593), the antibiotic treatment with DCA and the NLRP3 inhibitor MCC950 group (ABX + DCA + MCC950, MCE, HY−12815), the antibiotic treatment with DCA and FXR agonist GW4064 group (ABX + DCA + GW4064, MCE, HY−50108), and the ABX + DCA + MCC950 + GW4064 group., , The therapeutic dosage of the medication was administered according to the manufacturer's product instructions.

Techniques: Activation Assay, Cell Differentiation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: Environmental Health Perspectives

Article Title: Evaluation of FXR Activity in Pollutants Identified in Sewage Sludge and Subsequent in Vitro and in Vivo Characterization of Metabolic Effects of Triphenyl Phosphate

doi: 10.1289/EHP15435

Figure Lengend Snippet: Identification of potential FXR-active pollutants in sewage sludge samples using an FXR pull-down assay. (A) Schematic diagram of the FXR pull-down assay. (B) The recoveries of the FXR agonist OCA “pulled down” by the FXR-LBD protein or control protein ( n = 3 ). (C) Volcano plot of the peak features identified by nontargeted analysis in pulled-down sewage sludge samples ( n = 8 ). The orange dots represent the features with fold change > 1 and p < 0.05 , mainly consisting of endogenous metabolites and natural products. The red dots represent the environmental pollutants identified at level 1 confidence, and detailed information of these pollutants are provided in Table S6. (D) The MS 2 spectrum of the identified representative pollutant (TPHP) in comparison with its standard. The MS 2 spectra of the other identified pollutants and their respective standards are provided in Figure S2. Data are represented as mean ± SEM . Comparisons of two groups were determined using Student’s t -test. The numeric data are provided in Excel Table S1 and S2. Note: AHTN, tonalid; ATBC, acetyl tributyl citrate; BHT-C, 3,5-Ditert-butyl-4-hydroxybenzaldehyde; BPF, 2,2′-bisphenol F; CAP, chloramphenicol; DEHP, bis(2-ethylhexyl) phthalate; EFV, efavirenz; FIP-S, fipronil sulfone; FLU, fludioxonil; FXR, farnesoid X receptor; FXR-LBD, ligand binding domain of FXR; GAX, galaxolidone; LTD, loratadine; MCZ, miconazole; OCA, obeticholic acid; OEA, oleoyl ethanolamide; PEA, palmitoyl ethanolamide; SEM, standard error of the mean; TCC, triclocarban; TEHP, tris(2-ethylhexyl) phosphate; TIBP, triisobutyl phosphate; TLS, triclosan; TPHP, triphenyl phosphate; UV329, octrizole. * p < 0.05 , ** p < 0.01 , *** p < 0.001 .

Article Snippet: The performance of the FXR protein affinity pull-down assay was evaluated by incubating 100 nM of the FXR agonist obeticholic acid (OCA) (T1789; TargetMol) with the 6His-FXR-LBD protein and HIS-select nickel magnetic agarose beads following the same procedure as mentioned above.

Techniques: Pull Down Assay, Control, Comparison, Ligand Binding Assay