Resveratrol and Its Natural Analogs Mitigate Immune Dysregulation and Oxidative Imbalance in the Endometriosis Niche Simulated in a Co-Culture System of Endometriotic Cells and Macrophages

<b>Background:</b> Inflammation and immune cell dysfunction are critical facilitators of endometriosis pathophysiology. Macrophages are renowned for stimulating lesion growth, vascularization, innervation, and pain generation. By combining macrophages and endometriotic cells, we determined if resveratrol and its natural analogs can target the immune dysregulation and oxidative imbalance in endometriosis. <b>Methods:</b> After treatment with compounds (5, 10, 25 µM), we evaluated the expression of key inflammatory and oxidative stress markers, cytokines release, and ROS production by applying q-PCR, ELISA, Cytometric Beads Array, and multiplexed fluorogenic staining and flow cytometry analysis with bioimaging. <b>Results:</b> The results showed that endometriosis-related macrophages treated with stilbenes have impaired expression of pro-inflammatory markers (<i>IL6</i>, <i>IL8</i>, <i>IL1B</i>, <i>TNF</i>, <i>CCL2</i>, <i>CXCL10</i>, <i>PTGS2</i>). The effect of resveratrol, pterostilbene, and piceatannol was observed, especially in reducing <i>IL1B</i>, <i>CCL2</i>, and <i>CXCL10</i> genes up to 3.5-, 5-, and 7.7-fold at 25 µM, respectively. Also, with piceatannol or polydatin exposure, the IL-6 decrease was noticeable. This study reported an antioxidant effect by reducing ROS-positive cells from 96% to 48% by pterostilbene. Results from flow cytometry correlated with the transcript activation of detoxification enzymes (<i>SOD</i>, <i>GPX</i>). <b>Conclusions:</b> Prospects for potential therapy based on regulating the immune microenvironment and reducing the accumulation of free radicals with stilbenes application were described in the article.