Estrogenic activity of a naringinase-treated extract of Sophora japonica cultivated in Egypt

Figures & data

Table 1.  Induction of β-galactosidase in the yeast two-hybrid assay expressing ERβ by the methanol extract and fractions from S. japonica seeds before and after naringinase treatment (NT).

Extract	β-Galactosidase activity (U)
	ERβ
	100 μg/mL	10 μg/mL
MeOH extract	129.6 ± 1.6	127.2 ± 8.6
NT MeOH extract	1021.2 ± 40.1**	340.5 ± 6.8*
CHCl3 fraction	170.5 ± 11.1*	130.0 ± 9.6
25% MeOH extract (fr. A)	30.2 ± 7.4	37.9 ± 33.1
50% MeOH extract (fr. B)	24.1 ± 2.5	17.8 ± 3.4
100% MeOH extract (fr. C)	45.2 ± 7.8	25.3 ± 4.1
NT 25% MeOH extract (fr. A-NT)	871.2 ± 50.8**	638.1 ± 44.7**
NT 50% MeOH extract (fr. B-NT)	1192.1 ± 129.9**	871.2 ± 50.8**
NT 100% MeOH extract (fr. C-NT)	531.5 ± 10.6**	280.8 ± 66.7*

Each value represents the mean ± SE of three independent experiments (n =  3). Asterisks denote significant differences from the control at p < 0.05 (*), p < 0.01 (**). In the yeast expressing ERβ: dimethylsulfoxide (DMSO) was 40.1 ± 1.6 (U) and 17β-estradiol was 695.0 ± 20.6 at concentration of 10⁻⁷ M. In the yeast expressing ERα (methanol extracts): DMSO was 40.0 ± 7.0 and 17β-estradiol was 786.8 ± 32.5 at a concentration of 10⁻⁷ M.

Figure 1.  Chemical structures of the isolated compounds from S. japonica seeds.

Table 2.  Induction of β-galactosidase in the yeast two-hybrid assay expressing ERα and ERβ by the isolated compounds from S. japonica.

Compound	β-Galactosidase activity (U)
	ERα			ERβ
	10^−4 M	10^−5 M	10^−6 M	10^−4 M	10^−5 M	10^−6 M
1	54.1 ± 16.5	71.6 ± 3.3	77.3 ± 4.1	56.2 ± 1.0	4.9 ± 5.9	39.2 ± 1.4
2	83.1 ± 5.9	80.5 ± 1.7	74.7 ± 5.9	184.2 ± 4.7*	58.8 ± 2.6	54.8 ± 4.9
3	73.3 ± 5.6	85.5 ± 9.0	83.4 ± 8.4	38.4 ± 5.8	37.9 ± 1.5	36.1 ± 1.5
4	99.7 ± 13.1	105.6 ± 1.9	109.9 ± 2.6	71.3 ± 24.5	90.5 ± 8.4	41.8 ± 6.21
5	66.1 ± 20.1	65.5 ± 4.8	55.6 ± 3.5	35.8 ± 2.5	31.2 ± 1.4	31.7 ± 1.3
6	117.2 ± 6.2	89.6 ± 3.7	71.5 ± 6.2	87.7 ± 3.4	88.9 ± 5.8	83.5 ± 4.1
7	79.2 ± 12.4	72.3 ± 5.5	56.2 ± 6.1	69.8 ± 6.1	75.2 ± 8.4	70.9 ± 7.4
8	76.8 ± 10.5	70.5 ± 9.5	110.3 ± 5.4	81.5 ± 9.1	58.1 ± 10.1	65.7 ± 4.1
9	55.1 ± 1.9	50.1 ± 1.7	48.5 ± 1.7	30.4 ± 1.7	29.2 ± 4.3	27.6 ± 3.9
10	132.4 ± 27.2	153.4 ± 29.8	152.1 ± 4.6	89.3 ± 23.3	57.6 ± 4.4	42.8 ± 2.2
11	228.6 ± 14.7**	70.3 ± 17.6	40.4 ± 23.4	2685.0 ± 145.4**	698.2 ± 38.8**	106.6 ± 11.2
12	390.3 ± 39.8**	73.4 ± 11.6	41.1 ± 12.5	3378.5 ± 284.3**	1956.2 ± 281.3**	526.2 ± 130.8**

Each value represents the mean ± SE of three independent experiments (n = 3). Asterisks denote significant differences from the control at p < 0.05 (*), p < 0.01 (**). In the yeast expressing ERβ: the negative control (DMSO) was 42.3 ± 3.0 and 17β-estradiol was 3121.7 ± 219.9 at a concentration of 10⁻⁷ M. In the yeast expressing ERα: DMSO was 63.6 ± 3.5 and 17β-estradiol was 1538.7 ± 44.4 at a concentration of 10⁻⁷ M.

Table 3.  Inhibitory effects of the isolated compounds from S. japonica on induction of β-galactosidase activity by 17β-estradiol in yeast two-hybrid assay (ERα and ERβ).

Compound	β-Galactosidase activity (% of control)
	ERα		ERβ
	10^−4 M	10^−5 M	10^−4 M	10^−5 M
1	93.5 ± 7.3	109.6 ± 3.1	94.9 ± 11.1	102.4 ± 1.1
2	81.8 ± 7.3*	112.5 ± 6.2	106.9 ± 6.8	94.8 ± 7.3
3	116.4 ± 10.2	117.2 ± 17.1	116.7 ± 7.1	102.2 ± 8.5
4	100.7 ± 2.2	91.6 ± 11.5	114.1 ± 5.4	121.4 ± 6.7
5	109.8 ± 2.8	122.5 ± 2.5	98.8 ± 6.5	105.2 ± 4.3
6	116.2 ± 6.8	97.8 ± 8.9	112.3 ± 6.3	120.8 ± 4.6
7	125.5 ± 2.2	117.6 ± 7.1	108.4 ± 5.5	99.6 ± 6.5
8	103.2 ± 1.5	111.1 ± 3.2	106.5 ± 4.3	100.5 ± 1.2
9	102.5 ± 10.2	103.1 ± 2.8	96.4 ± 0.8	96.1 ± 6.8
10	85.7 ± 3.5*	114.9 ± 3.1	108.6 ± 5.6	94.3 ± 3.2
11	85.7 ± 3.2*	114.9 ± 9.7	116.2 ± 4.5	111.8 ± 9.7
12	81.0 ± 7.6*	93.1 ± 2.2	93.4 ± 1.5	106.4 ± 7.6
Tamoxifen	28.2 ± 1.5**	91.9 ± 3.2	22.4 ± 2.5**	98.8 ± 10.9

β-Galactosidase activity (U) of 17β-estradiol was 483.1 ± 22.5 and 2333.4 ± 112.5 in yeast expressing ERα and ERβ, respectively, at a concentration of 10⁻⁷ M (considered as 100% activity). β-Galactosidase activity (U) of the tested compounds was calculated as a percentage of the 17β-estradiol activity (control). Asterisks indicate significant differences from the control at p < 0.05 (*), p < 0.01 (**) (n = 3).