Mechanism Analysis of UCP2 During the Oxidative Stress Injury of Intestinal Porcine Epithelial Cell Line-J2

TL;DR

This study shows that UCP2 protects pig gut cells from oxidative stress, and genipin can worsen damage when stress is present.

Contribution

The study reveals UCP2's protective role in pig intestinal cells and how genipin modulates its effects under oxidative stress.

Findings

UCP2 overexpression improved cell survival and reduced oxidative damage in pig intestinal cells.

Genipin worsened oxidative stress effects by inhibiting UCP2 and increasing cell death markers.

UCP2 modulates antioxidant enzymes and apoptosis-related genes to protect cells.

Abstract

Modern pig farming often faces challenges caused by oxidative stress, which happens when harmful molecules called reactive oxygen species build up in the body. This stress can damage the gut, reduce nutrient absorption, and harm overall pig health. In this study, we explored how a natural protein found in cells, called uncoupling protein 2, helps protect pig intestinal cells from this damage. We increased the levels of this protein in pig gut cells grown in the lab and found that the cells were better able to survive oxidative stress. They produced fewer harmful molecules and were less likely to activate genes that cause cell death. We also tested genipin, a plant-derived compound that can reduce the levels of this protective protein. We discovered that while genipin had little effect under normal conditions, it worsened the damage when the cells were under oxidative stress. These results show that uncoupling protein 2 plays an important role in defending pig gut cells from stress and that targeting this protein may help develop new strategies to improve gut health, support growth, and increase disease resistance in pigs—contributing to more sustainable and healthy livestock farming.

Oxidative stress poses a significant challenge in livestock production, impairing intestinal function, nutrient absorption, and overall animal performance. Uncoupling protein 2 (UCP2) is a mitochondrial regulator known for its protective effects against oxidative damage, but its specific function in porcine intestinal epithelial cells and its regulation by genipin—a natural UCP2 inhibitor with potential therapeutic properties—remains unclear. In this study, we cloned and overexpressed the porcine UCP2 gene in intestinal porcine epithelial cells (IPEC-J2), generating a stable UCP2-overexpressing cell line (IPEC-J2-UCP2). Under hydrogen peroxide-induced oxidative stress, UCP2 overexpression significantly improved cell viability, reduced reactive oxygen species (ROS) levels, and enhanced antioxidant enzyme activities (SOD, GPx, and CAT). Additionally, UCP2 upregulated the anti-apoptotic gene Bcl-2 and downregulated pro-apoptotic genes (Fas, Caspase-3, and Bax), indicating a protective role against oxidative stress-induced apoptosis. We also investigated the regulatory effects of genipin on UCP2. Under non-stress conditions, genipin mildly promoted anti-apoptotic gene expression. However, under oxidative stress, genipin strongly inhibited UCP2 expression, exacerbated ROS accumulation, reduced cell viability, and increased expression of pro-apoptotic markers, particularly Caspase-3 and Bax. These findings reveal that UCP2 plays a critical role in protecting porcine intestinal epithelial cells from oxidative injury and that genipin exerts context-dependent effects on cell fate by modulating UCP2. This study provides a mechanistic basis for targeting UCP2 to manage oxidative stress and improve intestinal health and performance in pigs.

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