FXR/Farnesoid X receptor (IC50 = 28μM); Microbial Metabolite

Tauro-α-muricholic acid (T-α-MCA) (IC50 = 28 μM) and tauro-β-muricholic acid (T-α-MCA) (IC50 = 28 μM) and tauro-β-muricholic acid (T-β-MCA)) (IC50 = 40 μM) are both known as efficient natural antagonists of FXR (Chiang and Ferrell, 2018). They can competitively inhibit the activation of FXR by other bile acids (Li et al., 2013). Next, we found that the sum of cecal T-α-MCA and T-β-MCA levels were significantly upregulated in the LD group compared with the NC group, and their levels were downregulated by NaB treatment. The change in sum of T-α-MCA and T-β-MCA levels in serum was contrary to change observed in the cecal contents (Fig. 4H) [2].

NaB administration changed bile acid and cholesterol transporters in the liver and ileum in LD-fed mice [2]
To determine the effect of the FXR signaling pathway on the CGS mouse model (Li et al., 2013), we detected the signaling molecules related to this pathway. In the current study, we examined the levels of liver Fxr (Nr1h4), Shp (Nr0b2) and Fgfr4 mRNA. We found that Fxr and Shp were downregulated in LD-fed mice and upregulated by NaB treatment, but the change was not significant. The mRNA expression of Fgfr4 was significantly reduced in the LD group and upregulated by NaB treatment. Next, we assessed the expression of several genes encoding catalytic enzymes for the synthesis of bile acids and regulated by the FXR-FGF-15/SHP-FGFR4 signaling pathway: cholesterol 7α-hydroxylase (Cyp7a1), sterol 12α-hydroxylase (Cyp8b1), sterol 27-hydroxylase (Cyp27a1) and oxysterol 7α-hydroxylase (Cyp7b1). We found that these genes were markedly decreased in LD-fed mice compared with NC mice and were further decreased by NaB treatment (Fig. 5A). It was suggested that the synthesis of primary bile acids was inhibited in LD-fed mice and that NaB treatment exacerbated this inhibition. Furthermore, we found that the mRNA expression of Fxr in the ileum was markedly decreased in LD-fed mice compared with NC mice and was significantly upregulated by NaB treatment. The mRNA expression of Fgf-15 and Shp was upregulated in LD-fed mice and significantly further upregulated with NaB treatment (Fig. 5B). The protein expression of FGF-15 in the ileum was assessed by immunofluorescence, and the results were consistent with the mRNA level (Fig. 5C). These results suggested that a high dosage of CA administered in the ileum could have led to ileum FXR activation in LD-fed mice (Chiang and Ferrell, 2018). However, the high contents of T-α-MCA and T-β-MCA in the LD group partially inhibited the activity of FXR. The inhibition of FXR was reduced, and the expression of ileum FXR was upregulated by NaB treatment, which may lead to full FXR activation.

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After sacrifice, liver and ileum samples were collected at postoperative week 20. The BA profiles are showed in Figure 4. The amount of GUDCA (1687.7 ± 2352.3 ng/ml vs. 16.8 ± 17.2 ng/ml, P = 0.049), T-α-MCA and T-β-MCA (1800.0 ± 1857.5 ng/ml vs. 15.5 ± 14.0 ng/ml, P = 0.011) in liver tissues in the IT group was more than that in the SH group, and the amount of α-MCA (26.8 ± 29.9 ng/ml vs. 535.1 ± 314.3 ng/ml, P < 0.001) was less than that in the SH group. In ileal tissue samples, the amount of GUDCA (12673.2 ± 7011.8 ng/ml vs. 62.2 ± 88.9 ng/ml, P < 0.001), T-α-MCA and T-β-MCA (15645.0 ± 17267.4 ng/ml vs. 111.0 ± 179.7 ng/ml, P = 0.016) in the IT group was more than that in the SH group (Figure 4) [1].

Bile acid profiles [1]
To identify changes of BA profiles after IT, 17 kinds of BAs, such as α-muricholic acid (α-MCA), β-muricholic acid (β-MCA), cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), hyodeoxycholic acid (HDCA), glycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), glycodeoxycholic acid (GDCA), glycoursodeoxycholic acid (GUDCA), lithocholic acid (LCA), tauro-α-muricholic acid (T-α-MCA), tauro-β-muricholic acid (T-β-MCA), taurocholic acid (TCA), turoursodeoxycholic acid (TDCA), taurohyodeoxycholic acid (THDCA) and tauroursodeoxycholic acid (TUDCA) were measured in samples.

[1]. Farnesoid X receptor is inhibited after ileum transposition in diabetic rats: its hypoglycemic effect. Int J Med Sci. 2023 Apr 2;20(5):595-605.

[2]. Sodium butyrate alleviates cholesterol gallstones by regulating bile acid metabolism. Eur J Pharmacol. 2021 Oct 5;908:174341.

Background: This study aimed to investigate changes in bile acid profiles and farnesol X receptor (FXR) status after ileal transposition (IT) and to elucidate its potential hypoglycemic mechanism. Methods: Twenty male diabetic rats were randomly divided into an IT group and a sham-operated group (SH group). Bile acid profiles were determined using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Glucose metabolism was monitored after oral administration of FXR inhibitors and agonists. The expression of key FXR target genes was also detected. Results: Plasma levels of β-mouse cholic acid (P = 0.047), taurine-α-mouse cholic acid, and taurine-β-mouse cholic acid (P < 0.001) were higher in the IT group than in the SH group, while taurine cholic acid (P = 0.049) and ursodeoxycholic acid (P = 0.030) levels were lower in the IT group than in the SH group. Inhibition of intestinal FXR improved glucose metabolism in the SH group. Administration of FXR agonists increased blood glucose levels in both groups. After sacrifice, the levels of glycosylursodeoxycholic acid, taurine-α-mouse cholic acid and taurine-β-mouse cholic acid in the liver and ileum were higher than those in the SH group (P < 0.05), while the level of α-mouse cholic acid in the liver tissue was lower than that in the SH group (P < 0.001). In addition, the expression of CYP7A1 mRNA (P < 0.001) and FGF15 mRNA (P = 0.001) in the IT group was significantly higher than that in the SH group, while the expression of PEPCK mRNA (P = 0.004), SREPB1c mRNA (P = 0.005) and SRB1 mRNA (P = 0.001) was significantly lower than that in the SH group. Conclusion: We found that there was significant heterogeneity in the bile acid profile after IT, and FXR activation may have an adverse effect on glucose metabolism. [1] Cholesterol overload and bile acid metabolism disorder play an important role in the occurrence of cholesterol stones (CGS). Short-chain fatty acids (SCFAs) regulate bile acid metabolism by modulating gut microbiota. However, the role of sodium butyrate (NaB) in targeting bile acids to alleviate cholelithiasis (CGS) and its mechanism remain unclear. This study showed that daily administration of 12 mg NaB for 8 consecutive weeks reduced the incidence of litholytic diet (LD)-induced gallstones from 100% to 25%. NaB modulates SCFA levels and improves gut microbiota. Gut microbiota remodeling alters bile acid composition and reduces the levels of taurine-α-mouse bile acid (T-α-MCA) and taurine-β-mouse bile acid (T-β-MCA) in the cecum, both of which are potent farnesol X receptor (FXR) antagonists. Quantitative real-time PCR analysis showed that NaB significantly increased the expression levels of FXR, fibroblast growth factor-15 (Fgf-15), and small heterodimer chaperone (Shp) mRNA in the ileum, thereby inhibiting bile acid synthesis. In addition, NaB enhances bile acid excretion by increasing the levels of hepatic multidrug resistance protein 2 (Mdr2) and bile salt export pump (Bsep) mRNA, and enhances intestinal reabsorption of bile acids by increasing the levels of ileal bile acid transporter (Ibat) mRNA. Furthermore, NaB reduces intestinal cholesterol absorption and inhibits hepatic cholesterol excretion, thereby reducing serum and bile cholesterol concentrations. Moreover, FXR antagonists can eliminate the protective effect of NaB. In summary, our results indicate that NaB alleviates CGS by regulating the FXR-FGF-15/SHP signaling pathway through the regulation of gut microbiota. [2]

C26H44NNAO7S

537.68

537.273

2260905-08-4

25613-05-2

137700104

White to off-white solid powder

4

7

7

36

897

11

C[C@H](CCC(=O)NCCS(=O)(=O)[O-])[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2[C@@H]([C@H]([C@H]4[C@@]3(CC[C@H](C4)O)C)O)O)C.[Na+]

NYXROOLWUZIWRB-BAMGEBLESA-M

InChI=1S/C26H45NO7S.Na/c1-15(4-7-21(29)27-12-13-35(32,33)34)17-5-6-18-22-19(9-11-25(17,18)2)26(3)10-8-16(28)14-20(26)23(30)24(22)31;/h15-20,22-24,28,30-31H,4-14H2,1-3H3,(H,27,29)(H,32,33,34);/q;+1/p-1/t15-,16-,17-,18+,19+,20+,22+,23+,24+,25-,26-;/m1./s1

sodium;2-[[(4R)-4-[(3R,5R,6S,7S,8S,9S,10R,13R,14S,17R)-3,6,7-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]ethanesulfonate

2260905-08-4; Tauro-alpha-muricholic acid sodium salt; 2-[[(3alpha,5beta,6beta,7alpha)-3,6,7-trihydroxy-24-oxocholan-24-yl]amino]-ethanesulfonicacid,monosodiumsalt; sodium;2-[[(4R)-4-[(3R,5R,6S,7S,8S,9S,10R,13R,14S,17R)-3,6,7-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]ethanesulfonate; Tauro-; A-muricholic acid (sodium); Tauro-a-muricholic Acid Sodium Salt; Tauro-?-muricholic acid sodium;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)