The effects of resveratrol on the expression of VEGF, TGF-β, and MMP-9 in endometrial stromal cells of women with endometriosis

Abstract

Resveratrol is a phytochemical with anti-angiogenic, anti-inflammatory, and antioxidant properties. The present study has evaluated the effect of resveratrol on the expression of vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β) and matrix metalloproteinase-9 (MMP-9) as factors related to endometriosis progression. Thirteen eutopic (EuESCs) and 8 ectopic (EESCs) endometrial stromal cells from women with endometriosis and 11 control endometrial stromal cells (CESCs) were treated with resveratrol (100 µM) for 6, 24 and 48 h. The gene and protein expression levels of VEGF, TGF-β, and MMP-9 were measured using real-time PCR and ELISA methods, respectively. Results showed that the basal gene and protein expression of VEGF and MMP-9 were higher in EESCs compared to EuESCs and CESCs (P < 0.01 to < 0.001 and P < 0.05 to < 0.01 respectively). Also, resveratrol treatment decreased the gene and protein expression of VEGF and MMP-9 in EuESCs, EESCs and CESCs (P < 0.05 to < 0.01 and P < 0.05 to < 0.01 respectively) and gene and protein expression of TGF-β in EESCs and EuESCs (P < 0.05 to < 0.01). The effect of resveratrol in reduction of VEGF gene expression was statistically more noticeable in EESCs compared to EuESCs and CESCs (P < 0.05). According to the findings, resveratrol may ameliorate endometriosis progression through reducing the expression of VEGF, TGF-β, and MMP-9 in endometrial stromal cells (ESCs).

Figure 1

The basal expression levels of VEGF, TGF-β and MMP-9 genes and proteins in ESCs. The basal expression of VEGF, TGF-β and MMP-9 genes and proteins were measured in EESCs (n = 8) and EuESCs (n = 13) from endometriotic women and CESCs from non-endometriotic controls (n = 11) by real-time PCR and ELISA. (a) The basal expression of VEGF gene, (b) The basal expression of VEGF protein, (c) The basal expression of TGF-β gene, (d) The basal expression of TGF-β protein, (e) The basal expression of MMP-9 gene, (f) The basal expression of MMP-9 protein. *P-value < 0.05, ** P-value < 0.01 and *** P-value < 0.001 by non-parametric tests. VEGF: Vascular endothelial growth factor, TGF-β: Transforming growth factor-β, MMP-9: Matrix metalloproteinase-9, ESCs: Endometrial stromal cells, EuESCs: Eutopic endometrial stromal cells, EESCs: Ectopic endometrial stromal cells, CESCs: Control endometrial stromal cells.

Figure 2

Resveratrol Decreased the Expression of VEGF Protein in EuESCs and EESCs. ESCs from endometriotic women (8 EESCs and 13 EuESCs) and non-endometriotic controls (11 CESCs) were cultured with or without of 100 µM resveratrol. After 6, 24, and 48 h, the protein expression of VEGF was examined using ELISA. (a) 6 hr (Res + vs Res-), (b) 24 hr (Res + vs Res-), (c) 48 hr (Res + vs Res-). *P-value < 0.05, ** P-value < 0.01 by non-parametric tests. VEGF: Vascular endothelial growth factor, EuESCs: Eutopic endometrial stromal cells, EESCs: Ectopic endometrial stromal cells, CESCs: Control endometrial stromal cells.

Figure 3

Resveratrol Decreased the Expression of TGF-β Protein in EuESCs and EESCs. ESCs from endometriotic women (8 EESCs and 13 EuESCs) and non-endometriotic controls (11 CESCs) were cultured with or without of 100 µM resveratrol. After 6, 24, and 48 h, the protein expression of TGF-β was examined using ELISA. (a) 6 hr (Res + vs Res-), (b) 24 hr (Res + vs Res-), (c) 48 hr (Res + vs Res-). *P-value < 0.05 by non-parametric tests. TGF-β: Transforming growth factor-β, EuESCs: Eutopic endometrial stromal cells, EESCs: Ectopic endometrial stromal cells, CESCs: Control endometrial stromal cells.

Figure 4

Resveratrol Decreased the Expression of MMP-9 Protein in all ESCs. ESCs from endometriotic women (8 EESCs and 13 EuESCs) and non-endometriotic controls (11 CESCs) were cultured with or without of 100 µM resveratrol. After 6, 24, and 48 h, the protein expression of MMP-9 was examined using ELISA. (a) 6 hr (Res + vs Res-), (b) 24 hr (Res + vs Res-), (c) 48 hr (Res + vs Res-). *P-value < 0.05 by non-parametric tests. MMP-9: Matrix metalloproteinase-9, EuESCs: Eutopic endometrial stromal cells, EESCs: Ectopic endometrial stromal cells, CESCs: Control endometrial stromal cells.