Abstract：
Porcine epidemic diarrhea virus (PEDV) belongs to the Alphacoronavirus genus within the Coronavirus family, causing severe watery diarrhea in piglets and resulting in significant economic losses. Medium-chain acyl-CoA dehydrogenase (ACADM) is an enzyme participating in lipid metabolism associated with metabolic diseases and pathogen infections. Nonetheless, the precise role of ACADM in regulating PEDV replication remains uncertain. In this study, we identified ACADM as the host binding partner of NSP4 via immunoprecipitation-mass spectrometry analysis. The interaction between ACADM and NSP4 was subsequently corroborated through coimmunoprecipitation and laser confocal microscopy. Following this, a notable upsurge in ACADM expression was observed during PEDV infection. ACADM overexpression effectively inhibited virus replication, whereas ACADM knockdown facilitated virus replication, suggesting ACADM has negative regulation effect on PEDV infection. Furthermore, we demonstrated fatty acid β-oxidation affected PEDV replication for the first time, inhibition of fatty acid β-oxidation reduced PEDV replication. ACADM decreased PEDV-induced β-oxidation to suppress PEDV replication. Mechanistically, ACADM reduced cellular free fatty acid levels and subsequent β-oxidation by hindering AMPK-mediated lipophagy. In summary, our results reveal that ACADM plays a negative regulatory role in PEDV replication by regulating lipid metabolism. The present study introduces a novel approach for the prevention and control of PEDV infection.
摘要：
猪流行性腹泻病毒（PEDV）属于冠状病毒家族中的Alphacoronavirus属，引起仔猪严重的水样腹泻，造成重大的经济损失。中链酰基辅酶A脱氢酶（ACADM）是一种参与与代谢性疾病和病原体感染相关的脂质代谢的酶。尽管如此，ACADM在调节PEDV复制中的确切作用仍不确定。在这项研究中,我们通过免疫沉淀-质谱(IP-MS)分析鉴定了ACADM为NSP4的宿主结合伴侣.随后通过共免疫沉淀和激光共聚焦显微镜证实了ACADM和NSP4之间的相互作用。在此之后,在PEDV感染期间观察到ACADM表达显著升高。ACADM过表达有效抑制病毒复制,而ACADM敲除促进病毒复制，提示ACADM对PEDV感染有负调节作用。此外，我们首次证明脂肪酸β-氧化影响PEDV的复制，抑制脂肪酸β-氧化减少PEDV复制。ACADM降低PEDV诱导的β-氧化以抑制PEDV复制。机械上,ACADM通过阻碍AMPK介导的吸脂性降低了细胞游离脂肪酸（FFA）水平和随后的β-氧化。总之,我们的结果表明，ACADM通过调节脂质代谢在PEDV复制中起负调节作用。本研究提出了一种预防和控制PEDV感染的新方法。
