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Background <p>Infectious bovine rhinotracheitis virus (IBRV), a member of the Herpesviridae family, causes infectious bovine rhinotracheitis (IBR) and induces mitochondrial dysfunction in host cells. Circular RNAs (circRNAs)—a novel class of non-coding RNAs—have been implicated in various biological processes and pathologies related to mitochondrial damage. However, their role in IBRV-induced mitochondrial damage in Madin-Darby bovine kidney (MDBK) cells remains unclear.</p> Results <p>Transmission electron microscopy(TEM), laser confocal microscopy, and flow cytometry confirmed that IBRV infection causes mitochondrial damage in MDBK cells. High-throughput sequencing revealed 144 differentially expressed (DE) circRNAs, 725 messenger RNAs (mRNAs), and 160 microRNAs (miRNAs) in IBRV-infected cells. We predicted that DE circRNAs regulate mitochondrial damage via source genes of circRNA, circRNA-miRNA-mRNA networks, and RNA-binding proteins (RBPs). Source genes of circRNA were enriched in mitochondria-related pathways, such as the mammalian target of rapamycin (mTOR), thyroid hormone, and Hippo signalling; 11 genes were localized to mitochondria. CircRNA-miRNA-mRNA network target genes were associated with cellular senescence, mitophagy, and ubiquitin-mediated proteolysis; 471 genes were linked to mitochondria. Additionally, 961 RBPs were enriched in pathways, such as nucleocytoplasmic transport and RNA degradation; 107 RBPs were localized to mitochondria. Functional validation revealed knockdown of circ_002584 reduced reactive oxygen species (ROS) accumulation (<i>p</i> &lt; 0.05) and mitochondrial membrane potential depolarization (<i>p</i> &lt; 0.05). Knockdown of circ_004326 increased both (<i>p</i> &lt; 0.01).</p> Conclusions <p>CircRNAs play a regulatory role in IBRV-induced mitochondrial damage within MDBK cells. This finding is significant for virus-associated mitochondrial damage research, forming a theoretical foundation for utilizing circRNAs as diagnostic biomarkers and potential therapeutic targets for IBR.</p>

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Identification and functional analysis of circular RNAs during mitochondrial damage induced by infectious bovine rhinotracheitis virus infection in Madin–Darby bovine kidney cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月24日

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  • Yingcai Ma,
  • Jiaxin Liu,
  • Lianping Xu,
  • Qin He,
  • Heqi Lv,
  • Zelong Li,
  • Na Li,
  • Yawei Sun,
  • Pengfei Yi,
  • Mengli Xu,
  • Xiaomin Zhao,
  • Qi Zhong,
  • Gang Yao,
  • Xuelian Ma

摘要

Background

Infectious bovine rhinotracheitis virus (IBRV), a member of the Herpesviridae family, causes infectious bovine rhinotracheitis (IBR) and induces mitochondrial dysfunction in host cells. Circular RNAs (circRNAs)—a novel class of non-coding RNAs—have been implicated in various biological processes and pathologies related to mitochondrial damage. However, their role in IBRV-induced mitochondrial damage in Madin-Darby bovine kidney (MDBK) cells remains unclear.

Results

Transmission electron microscopy(TEM), laser confocal microscopy, and flow cytometry confirmed that IBRV infection causes mitochondrial damage in MDBK cells. High-throughput sequencing revealed 144 differentially expressed (DE) circRNAs, 725 messenger RNAs (mRNAs), and 160 microRNAs (miRNAs) in IBRV-infected cells. We predicted that DE circRNAs regulate mitochondrial damage via source genes of circRNA, circRNA-miRNA-mRNA networks, and RNA-binding proteins (RBPs). Source genes of circRNA were enriched in mitochondria-related pathways, such as the mammalian target of rapamycin (mTOR), thyroid hormone, and Hippo signalling; 11 genes were localized to mitochondria. CircRNA-miRNA-mRNA network target genes were associated with cellular senescence, mitophagy, and ubiquitin-mediated proteolysis; 471 genes were linked to mitochondria. Additionally, 961 RBPs were enriched in pathways, such as nucleocytoplasmic transport and RNA degradation; 107 RBPs were localized to mitochondria. Functional validation revealed knockdown of circ_002584 reduced reactive oxygen species (ROS) accumulation (p < 0.05) and mitochondrial membrane potential depolarization (p < 0.05). Knockdown of circ_004326 increased both (p < 0.01).

Conclusions

CircRNAs play a regulatory role in IBRV-induced mitochondrial damage within MDBK cells. This finding is significant for virus-associated mitochondrial damage research, forming a theoretical foundation for utilizing circRNAs as diagnostic biomarkers and potential therapeutic targets for IBR.