Table 3. The effects of coumestrol exposure on mRNA expressions of Th- and Treg-specific transcription factors and typical cytokines in the spleens of female CBA/J mice after induction of EAT.

		Control group	L group	H group
Th1	T-bet	0.82 ± 0.19	0.60 ± 0.19	0.79 ± 0.20
	IFN-γ	4.86 ± 0.77	1.42 ± 1.19 *	3.54 ± 1.62
Th2	GATA3	0.76 ± 0.10	1.02 ± 0.75	0.77 ± 0.24
	IL-4	1.85 ± 0.41	2.47 ± 0.38	2.20 ± 1.33
Th17	ROR-γt	0.17 ± 0.03	0.20 ± 0.08	0.19 ± 0.11
	IL-17	0.12 ± 0.08	0.21 ± 0.08	0.14 ± 0.08
Treg	Foxp3	5.47 ± 1.82	3.91 ± 1.96	4.45 ± 1.22
	TGF-β	0.12 ± 0.04	0.07 ± 0.02	0.11 ± 0.06

Female adult CBA/J mice were exposed to drinking water containing 1% Tween80 only (Control group), 0.8 mg/l coumestrol in 1% Tween80 (L group), 8 mg/l coumestrol in 1% Tween80 (H group), respectively. All the three groups of mice were further immunized with mouse thyroglobulin and Freund's adjuvant for induction of EAT. The mRNA expressions of Th- and Treg-specific transcription factors and typical cytokines in the spleens were examined by quantitative RT-PCR. The transcript levels were assessed using the comparative Ct method, normalized to that of murine GAPDH, and expressed as mean ±SD of △Ct values (n=5-8/group). Comparisons between groups were performed using one-way ANOVA followed by Bonferroni's post hoc test. *P <0.05 vs the control group.