Constitutive androstane receptor agonist, TCPOBOP, attenuates steatohepatitis in the methionine choline-deficient diet-fed mouse

Table 1 Primer sequences used for RT-PCR

Target	Primer sequence	Product size (bp)
ACO	F 5'-GAACTCCAGATAATTGGCACCTA-3'	75
	R 5'-AGTGGTTTCCAAGCCTCGAA-3'
CPT-1b	F 5'-ATCATGTATCGCCGCAAACT-3'	85
	R 5'-CCATCTGGTAGGAGCACATGG-3'
CYP2B10	F 5'-CAA TGGGGA ACG TTG GAA GA-3'	176
	R 5'-TGATGCACTGGAAGAGGA AC-3'
CYP3A11	F 5'-CTCAATGGTGTGTATATCCCC-3'	423
	R 5'-CCGATGTTCTTAGACACTGCC-3'
CYP4A10	F 5'-AGTGTCTCTGCTCTAAGCC-3'	180
	R 5'-CCCAAAGAACCAGTGAAA-3'
Ketoacyl thiolase	F 5'-GCATCCCAGAGACTGTACCTTT-3'	202
	R 5'-GTCTCTGGCCTTCTCGTTCT-3'
L-PBE	F 5'-TGGCTCTTGGAGGAGGACTAG A-3'	125
	R 5'-AAGCTGCGTTCCTCTTGCA-3'
MPT-α	F 5'-GGCAGTCTCAGTCGCTTCTC-3'	240
	R 5'-GCACTCCTGATTTGGTCGTT-3'
MPT-β	F 5'-AGAGCTGCACTTTCGGGTTT-3'	202
	R 5'-CTGTGGTCATGGCTTGGTTT-3'
PPAR-α	F 5'-GTACGGTGT GTATGAAGCCATCTT-3'	76
	R 5'-GCCGTACGCGATCAGCAT-3'
Urate oxidase	F 5'-ACCTCCCGTCATTCACTCT-3'	438
	R 5'-ACTGTCCCTGTTATTTTGCC-3'

ACO: Acetyl-CoA oxidase; CPT1: Carnitine palmitoyltransferase; Ketoacyl thiolase: Peroxisomal 3-ketoacyl-CoA thiolase; L-PBE: L-peroxisomal bifunctional enzyme; MTP: Mitochondrial trifunctional protein subunits alpha and beta; PPAR-α: Peroxisome proliferators-activated receptor-alpha.

Table 2 Effect of TCPOBOP on genes involved in fatty acid oxidation

Type of fatty acid oxidation	Gene	MCD	MCD-TCPOBOPa	Fold-change	P value
ω-oxidation	CYP4A10	1.40 ± 1.69	6.07 ± 1.73	4.3	0.001
(ER)	CYP4A14	0.81 ± 0.04	0.07 ± 0.02	-11.6	0.008
β-oxidation	L-PBE	0.07 ± 0.01	0.39 ± 0.01	5.6	0.0004
(peroxisome)	Ketoacyl thiolase	0.12 ± 0.03	0.28 ± 0.06	2.3	0.04
	ACO	0.25 ± 0.14	0.32 ± 0.08	1.3	NS
	Urate oxidase	0.29 ± 0.02	0.16 ± 0.02	-1.8	NS
β-oxidation	CPT1	0.79 ± 0.21	0.77 ± 0.19	-1.0	NS
(mitochondria)	MTP-α	0.60 ± 0.30	0.88 ± 0.71	1.5	NS
	MTP-β	0.50 ± 0.40	0.59 ± 0.07	1.2	NS

Values are expressed as mRNA expression/18S ratio ± SD.

^aAnimals fed methionine choline-deficient (MCD) diet for 2 wk and pre-treated with the CAR agonist, TCPOBOP (3 mg/kg ip for 3 d at starting of diet). NS: Not significant; ER: Endoplasmic reticulum; L-PBE: L-peroxisomal bifunctional enzyme; ACO; Acetyl-CoA oxidase; Ketoacyl thiolase: Peroxisomal 3-ketoacyl-CoA thiolase; CPT1: Carnitine palmitoyltransferase; MTP: Mitochondrial trifunctional protein subunits alpha and beta.