College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China.
Institute of Nutritional and Metabolic Disorders in Domestic Animals and Fowls, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China.
Vet Res. 2018 Feb 13;49(1):15. doi: 10.1186/s13567-018-0508-z.
Porcine circovirus type 2 (PCV2) is recognized as the causative agent of porcine circovirus-associated diseases. PCV2 replication could be promoted by low doses of ochratoxin A (OTA) as in our previous study and selenium has been shown to attenuate PCV2 replication. However, the underlying mechanism remains unclear. The aim of the study was to investigate the effects of selenomethionine (SeMet), the major component of organic selenium, on OTA-induced PCV2 replication promotion and its potential mechanism. The present study demonstrates that OTA could promote PCV2 replication as measured by cap protein expression, viral titer, viral DNA copies and the number of infected cells. In addition, OTA could activate autophagy as indicated by up-regulated light chain 3 (LC3)-II and autophagy-related protein 5 expressions and autophagosome formation. Further, OTA could down-regulate p-AKT and p-mTOR expressions and OTA-induced autophagy was inhibited when insulin was applied. SeMet at 2, 4 and 6 μM had significant inhibiting effects against OTA-induced PCV2 replication promotion. Furthermore, SeMet could attenuate OTA-induced autophagy and up-regulate OTA-induced p-AKT and p-mTOR expression inhibition. Rapamycin, an inhibitor of AKT/mTOR, could reverse the effects of SeMet on OTA-induced autophagy and the PCV2 replication promotion. In conclusion, SeMet could block OTA-induced PCV2 replication promotion by inhibiting autophagy by activating the AKT/mTOR pathway. Therefore, SeMet supplementation could be an effective prophylactic strategy against PCV2 infections and autophagy may be a potential marker to develop novel anti-PCV2 drugs.
猪圆环病毒 2 型(PCV2)被认为是猪圆环病毒相关疾病的病原体。在我们之前的研究中发现,低剂量的赭曲霉毒素 A(OTA)可以促进 PCV2 的复制,而硒已被证明可以抑制 PCV2 的复制。然而,其潜在机制尚不清楚。本研究旨在探讨硒蛋氨酸(SeMet),即有机硒的主要成分,对 OTA 诱导的 PCV2 复制促进及其潜在机制的影响。本研究表明,OTA 可以通过衣壳蛋白表达、病毒滴度、病毒 DNA 拷贝数和感染细胞数量来促进 PCV2 的复制。此外,OTA 可以激活自噬,表现为 LC3-II 和自噬相关蛋白 5 的表达上调和自噬体形成。此外,OTA 可以下调 p-AKT 和 p-mTOR 的表达,当应用胰岛素时,OTA 诱导的自噬受到抑制。2、4 和 6 μM 的 SeMet 对 OTA 诱导的 PCV2 复制促进具有显著的抑制作用。此外,SeMet 可以减弱 OTA 诱导的自噬,并上调 OTA 诱导的 p-AKT 和 p-mTOR 表达抑制。雷帕霉素,AKT/mTOR 的抑制剂,可以逆转 SeMet 对 OTA 诱导的自噬和 PCV2 复制促进的作用。总之,SeMet 通过激活 AKT/mTOR 通路抑制自噬来阻断 OTA 诱导的 PCV2 复制促进。因此,补充 SeMet 可能是预防 PCV2 感染的有效策略,自噬可能是开发新型抗 PCV2 药物的潜在标志物。