Aberrant TGF-β1 signaling contributes to the development of primary biliary cirrhosis in murine model

Table 1 Primers for real-time polymerase chain reaction

Gene	Genbank no	Forward primer (5' to 3')	Reverse primer (5' to 3')
TGFβ1	NM_011577	TGCTAATGGTGGACCGCAA	CACTGCTTCCCGAATGTCTGA
TβRI	NM_009370	ATGGTTCCGAGAGGCAGAGAT	CCATGTCCCATTGTCTTTGTTG
TβRII	NM_009371	CCAGAAGTCCTGCATGAGCAA	TGGCAAACCGTCTCCAGAGTA
Smad2	NM_010754	TCTCCGGCTGAACTGTCTCCTA	GCGATTGAACACCAGAATGCA
Smad3	NM_016769	ATGGAGCTCTGTGAGTTTGCCT	TGGAGGTAGAACTGGCGTCTCT
α-SMA	NM_007392	CTATTCAGGCTGTGCTGTCCCT	GCCCTCATAGATAGGCACGTTG
α1(I) collagen	NM_007742	CCCAAGGAAAAGAAGCACGTC	AGGTCAGCTGGATAGCGACATC
GAPDH	NM_008084	AGCCTCGTCCCGTAGACAAAA	TGGCAACAATCTCCACTTTGC

TGF: Transforming growth factor; TβR: TGF-β receptor; SMA: Smooth muscle actin; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Table 2 mRNA levels of transforming growth factor-β1 transforming growth factor-β receptorI, transforming growth factor-β receptorII, Smad2, Smad3, α-smooth muscle actin and α1 (I) collagen in mouse model and control mice

	Control mice	Mouse model
		week 8	week 24
TGF-β1	1.7 ± 0.4	7.0 ± 1.8b	8.9 ± 1.8b
TβRI	0.8 ± 0.2	2.8 ± 0.7b	5.1 ± 1.5b
TβRII	0.6 ± 0.01	1.9 ± 0.9b	5.1 ± 0.1b
Smad2	0.6 ± 0.3	3.8 ± 1.1b	2.0 ± 0.3b
Smad3	0.9 ± 0.4	1.7 ± 0.8a	3.4 ± 0.6b
α-SMA	0.8 ± 0.4	1.8 ± 0.1a	1.7 ± 0.3b
α1 (I) collagen	1.1 ± 1.2	11.0 ± 1.5b	11.8 ± 0.6b

The mRNA fold changes were calculated using glyceraldehyde-3-phosphate dehydrogenase as a control. Values were expressed as mean ± SD from 3 independent experiments.

^aP < 0.05,

^bP < 0.01 vs control mice. TGF: Transforming growth factor; TβR: TGF-β receptor; SMA: Smooth muscle actin.

Table 3 Phenotype of mononuclear cells in the liver

	Mouse model	Control mice
Total cell number (× 106)	1.6 ± 0.47a	0.48 ± 0.32
CD4+ (× 106)	0.06 ± 0.01	0.04 ± 0.01
CD8+ (× 106)	0.58 ± 0.11a	0.08 ± 0.03
CD4+/CD8+	0.12 ± 0.04a	0.52 ± 0.23
CD4+ CD25+ FOXP3+ (× 106)	0.01 ± 0.001b	0.004 ± 0.001

^aP < 0.05,

^bP < 0.01 vs control mice.