AMPK-PINK1/Parkin Mediated Mitophagy Is Necessary for Alleviating Oxidative Stress-Induced Intestinal Epithelial Barrier Damage and Mitochondrial Energy Metabolism Dysfunction in IPEC-J2.

Shuting Cao,Li Wang,Jingchun Gao,Jiang Zhu,Caihong Hu,Xin Li,Hao Xiao

doi:10.3390/antiox10122010

Abstract

The imbalance of redox biology and oxidative stress leads to intestinal barrier injury and mitophagy. However, much uncertainty still exists about the role of mitophagy in oxidative stress and intestinal function. Here, we showed the effects of hydrogen peroxide (H2O2)-induced oxidative stress on intestinal epithelial cell oxidation balance, intestinal barrier function and mitochondrial energy metabolism and its underlying mechanism. In this study, we found that H2O2-induced oxidative stress activated adenosine monophosphate-activated protein kinase (AMPK) and enhanced mitophagy in intestinal porcine epithelial cells (IPEC-J2). While compound C (AMPK inhibitor) and mdivi-1 (mitophagy inhibitor) significantly reduced the activity of superoxide dismutase (SOD) and increased mitochondrial reactive oxygen species (ROS) levels in H2O2 treated cells. Moreover, compound C and mdivi-1 significantly reduced the trans-epithelium electrical resistant (TER) and increased the fluorescein isothiocyanate-dextran (FD4) flux in H2O2 treated IPEC-J2. Furthermore, compound C and mdivi-1 significantly reduced the activity of mitochondrial complex II. Seahorse XF96 data showed that compound C + mdivi-1+ H2O2 treatment significantly reduced maximum respiratory oxygen consumption and spare respiratory capacity. Additionally, compound C or mdivi-1 treatment reduced the formation of mitochondrial autophagosomes. These results unveiled that AMPK and PINK1/Parkin mediated mitophagy is necessary for alleviating oxidative stress induced intestinal epithelial barrier damage and mitochondrial energy metabolism dysfunction in IPEC-J2.

Highlights

Weaning stress is extremely serious during the initial post-weaning period, which is accompanied by intestinal injury, diarrhea and even death, impairing the growth performance and anti-disease ability of piglets and causing great economic losses to the pig industry [1,2,3]
Several studies reported that imbalance redox biology and oxidative stress led to intestinal function injury [8,9]
The results showed that treated IPEC-J2 with 600 μM, 800 μM and 1000 μM H2 O2 for 8 h significantly reduced cell viability and 600 μM H2 O2 was used in this study to set an oxidative stress model

Summary

Introduction

Weaning stress is extremely serious during the initial post-weaning period, which is accompanied by intestinal injury, diarrhea and even death, impairing the growth performance and anti-disease ability of piglets and causing great economic losses to the pig industry [1,2,3]. Our previous studies and others reported that weaning impaired free-radical metabolism and the antioxidative system and resulted in serious oxidative stress [4,5,6]. Oxidative stress was interpreted as an imbalance between oxidation and anti-oxidation, resulting in the accumulation of reactive oxygen species (ROS) [7]. Several studies reported that imbalance redox biology and oxidative stress led to intestinal function injury [8,9]. Much uncertainty still exists about the underlying mechanism between oxidative stress and intestinal function.

Methods

Results

Conclusion