Anti‑inflammatory and anti‑migratory properties of wild‑grape‑derived ε‑viniferin in human endometrial cells: A potential therapy for endometriosis

Abstract

Endometriosis is a chronic gynecological disorder characterized by the ectopic growth and proliferation of endometrial tissue outside the uterine cavity, leading to inflammation. Although low‑dose contraceptive pills are widely used for its treatment, these therapies are associated with side effects and are contraindicated in women trying to conceive. ε‑viniferin is a resveratrol dimer found in wild grapes (Ampelopsis brevipedunculata) with demonstrated antioxidant properties and an anti‑migratory effect in lung cancer cells. Because abnormal cell migration is a key process in endometriosis, we hypothesized that ε‑viniferin could also exhibit anti‑migratory effects in endometriotic cells, which remain to be elucidated. To test this hypothesis, the present study investigated the anti‑inflammatory and anti‑endometriotic effects of ε‑viniferin and wild‑grape extract. RAW264.7 and THP‑1 were used to evaluate the anti‑inflammatory effects of wild‑grape extract and ε‑viniferin by assessing cytotoxicity; nitric oxide (NO), reactive oxygen species and interleukin‑6 (IL‑6) production; and NF‑κB activity. Additionally, human endometrial stromal cells (HESCs) were used to investigate the anti‑endometriotic effects of ε‑viniferin, including its impact on cell migration, invasion and gene expression, using PCR array analysis. Both ε‑viniferin and wild‑grape extract significantly reduced lipopolysaccharide‑induced NO and IL‑6 production, indicating an anti‑inflammatory activity. Inhibition of NF‑κB activity at the cellular level further supported these findings. Moreover, both ε‑viniferin and wild‑grape extract effectively suppressed the migration and invasion of HESCs, indicating their potential to alleviate endometriosis symptoms. These findings suggest that ε‑viniferin is a promising therapeutic candidate for endometriosis, exhibiting both anti‑inflammatory and anti‑migratory effects. Our results present a novel approach for developing effective anti‑endometriotic therapies.

Keywords: endometriosis; inflammation; invasion; migration; wild‑grape exact; ε‑viniferin.

Figure 1.

Chemical structure of ε-viniferin.

Figure 2.

Inhibition of inflammatory regulators and ROS production by wild-grape extract in RAW264.7 cells. (A) Effect of wild-grape extract on cell viability after 24 h of exposure, assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay. (B) Inhibition of LPS-induced NO production. (C) Reduction of LPS-induced ROS production, measured based on fluorescent intensity. (D) In vitro antioxidant activity determined using the DPPH radical-scavenging assay. (E) Effect on LPS-induced expression of iNOS, IL-6 and IL-1β. (F) Inhibition of LPS-induced IL-6 expression. (G) Inhibition of LPS-induced IL-6 secretion. RAW264.7 cells were stimulated with 100 ng/ml LPS in the presence or absence of wild-grape extract. n=3. *P<0.05, **P<0.01, ***P<0.001 vs. control without the extract. LPS, lipopolysaccharide; NO, nitric oxide; ROS, reactive oxygen species; DPPH, 2,2-diphenyl-1-picrylhydrazyl; iNOS, inducible NOS; IL, interleukin; mGapdh, mouse glyceraldehyde-3-phosphate dehydrogenase.

Figure 3.

Inhibition of inflammatory regulator production and NF-κB activity by ε-viniferin in RAW264.7 cells. (A) Effect of ε-viniferin on cell viability after 24 h of exposure. (B) In vitro antioxidant activity determined using the DPPH radical-scavenging assay. (C) Inhibition of LPS-induced NO production. (D) Inhibition of LPS-induced ROS production, measured based on fluorescent intensity. (E) Effect of ε-viniferin on cell viability after 4 h of exposure. (F) Inhibition of LPS-induced IL-6 expression. (G) Inhibition of LPS-induced IL-6 secretion. (H) Inhibition of LPS-induced NF-κB activity. RAW264.7 cells were stimulated with 100 ng/ml LPS in the presence or absence of ε-viniferin. N=3. *P<0.05, **P<0.01, ***P<0.001 vs. control without ε-viniferin and LPS. LPS, lipopolysaccharide; NO, nitric oxide; ROS, reactive oxygen species; IL, interleukin; DPPH, 2,2-diphenyl-1-picrylhydrazyl; mGapdh, mouse glyceraldehyde-3-phosphate dehydrogenase.

Figure 4.

Inhibition of IL-6 expression and secretion by ε-viniferin in THP-1 cells. (A) Effect of ε-viniferin on cell viability after 4 h of exposure. (B) Inhibition of LPS-induced IL-6 expression. (C) Inhibition of LPS-induced IL-6 secretion. THP-1 cells were stimulated with 100 ng/ml LPS in the presence or absence of ε-viniferin. N=3. *P<0.05, **P<0.01, ***P<0.001 vs. control without ε-viniferin and LPS. IL, interleukin; LPS, lipopolysaccharide. ACTB, β-actin.

Figure 5.

Inhibition of migration and invasion of human endometrial stromal cells by ε-viniferin. (A) Effect of ε-viniferin on cell viability after 24 h of exposure. (B) Inhibition of cell migration assessed using the wound healing assay after 8 h of treatment. (C) Suppression of cell invasion measured using the Matrigel invasion chamber assay after 16 h of treatment. Magnification, ×100. N=3. **P<0.01, ***P<0.001 vs. control without ε-viniferin.