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Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy
Published on: May 4, 2016
Porcine epidemic diarrhea virus induces Vero cell mitophagy by regulating ROS-mediated PINK1/Parkin signal pathway
1College of Animal Science and Technology, Inner Mongolia Minzu University, Tongliao, China.
Introduction:
Porcine epidemic diarrhea (PED) is a highly contagious enteric disease caused by the porcine epidemic diarrhea virus (PEDV), with mortality rates exceeding 80% in piglets. While the pathogenesis of PEDV is well-documented, its relationship with mitophagy remains unclear.
Methods:
We used transmission electron microscopy (TEM) to observe ultrastructural changes in PEDV-infected Vero cells. Western blotting and biochemical assays were employed to analyze mitophagy-related markers (LC3-II, PINK1, Parkin, Beclin1) and oxidative stress indicators (ROS, MDA, antioxidant enzymes). The role of the PINK1/Parkin pathway was further validated through siRNA knockdown and overexpression.
Results:
PEDV infection was found to trigger PINK1/Parkin-dependent mitophagy, evidenced by the accumulation of damaged mitochondria and upregulation of mitophagy markers. This process was driven by ROS-induced oxidative stress, as ROS scavenging significantly attenuated mitophagy. Furthermore, we demonstrated that mitophagy facilitates PEDV replication, with Parkin expression acting as a critical regulator of this viral-induced autophagic pathway.
Discussion:
This study reveals that PEDV exploits PINK1/Parkin-mediated mitophagy via ROS accumulation to promote viral replication. These findings provide a scientific basis for understanding PEDV pathogenesis and offer a technical reference for optimizing high-titer virus production in vaccine development.
