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3 protocols using taurohyocholic acid

1

Quantitative Bile Acid Analysis

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Optima grade acetonitrile and water were purchased from Fisher Scientific (Loughborough, UK). Ammonium acetate was purchased from Sigma-Aldrich (Dorset, UK). Formic acid was purchased from Fluka (Buchs, Switzerland). Bile acid standards, including tauro-alpha-muricholic aicd (T-α-MCA), tauro-beta-muricholic acid (T-β-MCA), tauro-omega-muricholic acid (T-ω-MCA), alpha-muricholic acid (α-MCA), beta-muricholic acid (β-MCA), taurohyocholic acid (THCA), glycoursodeoxycholic acid (GUDCA), tauroursodeoxycholic acid (TUDCA), hyocholic acid (HCA), taurohyoxydeoxycholic acid (THDCA), taurocholic acid (TCA), cholic acid (CA), ursodeoxycholic acid (UDCA), hyodeoxycholic acid (HDCA), glycodeoxycholic acid (GDCA), taurodeoxycholic acid (TDCA), chenodeoxycholic acid (CDCA), and deoxycholic acid (DCA) were purchased from Steraloids, Inc. (Newport, RI). Standard solutions of these 20 bile acids were prepared at concentrations of 0.5, 10, 50, 100, 500, 1500, 5000, and 10 000 nM in a solution of 20% (v/v) acetonitrile and water.
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2

Bile Acid Profiling with LC-MS

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Optima grade acetonitrile and water were purchased from Fisher Scientific (Loughborough, UK). Ammonium acetate was purchased from Sigma-Aldrich (Dorset, UK). Formic acid was purchased from Fluka (Buchs, Switzerland). Bile acid standards were purchased from Steraloids, Inc. (Newport, RI) and included tauro-alpha-muricholic acid (T-α-MCA), tauro-beta-muricholic acid (T-β-MCA), tauro-omega-muricholic acid (T-ω-MCA), alpha-muricholic acid (α-MCA), beta-muricholic acid (β-MCA), taurohyocholic acid (THCA), glycoursodeoxycholic acid (GUDCA), tauroursodeoxycholic acid (TUDCA), hyocholic acid (HCA), taurohyoxydeoxycholic acid (THDCA), taurocholic acid (TCA), cholic acid (CA), ursodeoxycholic acid (UDCA), hyodeoxycholic acid (HDCA), glycodeoxycholic acid (GDCA), taurodeoxycholic acid (TDCA), chenodeoxycholic acid (CDCA), and deoxycholic acid (DCA).
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3

Baicalin and Bile Acid Regulation

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Baicalin (>98%) was purchased from Dalian Meilun Biotechnology Co., Ltd (Meilunbio, Dalian, China). EE was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Standards for BAs for cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), β-muricholic acid (β-MCA), taurocholic acid (TCA), taurodeoxycholic acid (TDCA), tauromuricholic acid (T-MCA), taurochenodeoxycholic acid (TCDCA), glycocholic acid (GCA), glycoursodesoxycholic acid (GUDCA), glycohyocholic acid (GHDCA), glycochenodeoxycholic acid (GCDCA), taurohyocholic acid (THDCA), glycodeoxycholic acid (GDCA), and interior label d4-glycochenodeoxycholic acid (d4-GCDCA) were obtained from Steraloids (Newport, RI, USA).
Antibodies directed against FXR, CAR, PXR, VDR, cholesterol 7a-hydroxylase (CYP7A1), and sterol 27-hydroxylase (CYP27A1) were purchased from Absin Biochemical Company (Shanghai, China). Antibodies directed against Sirt1, HNF-1α, tumor necrosis factor α (TNF-α), interleukin 1 beta (IL-1β), IL-6, and NF-κB were purchased from Cell Signaling Technology (Beverly, MA, USA). An antibody against BSEP was obtained from Santa Cruz Biotechnology (CA, USA). And an antibody against MRP2 was obtained from Abcam (Cambridge, UK).
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