Ochratoxin A-induced autophagy in vitro and in vivo promotes porcine circovirus type 2 replication.
Ochratoxin A-induced autophagy in vitro and in vivo promotes porcine circovirus type 2 replication.
复制标题
赭曲霉毒素 A 诱导的体外和体内自噬促进猪圆环病毒 2 型复制。
DOI：
10.1038/cddis.2017.303
复制
发表时间：
2017-06-29
影响因子：
9
通讯作者：
Huang K
中科院分区：
文献类型：
--
作者：
Qian G;Liu D;Hu J;Gan F;Hou L;Chen X;Huang K
赭曲霉毒素A（OTA）是由曲霉属和青霉属产生的一种真菌毒素。猪圆环病毒2型（Porcine Circovirus type 2，PCV 2）是引起猪圆环病毒相关性疾病的病原体。最近，我们报道了低剂量的OTA在体外和体内促进PCV 2的复制，但其潜在的机制需要进一步研究。本研究进一步证实OTA诱导的PCV 2复制促进，如通过cap蛋白表达、病毒滴度、病毒DNA拷贝和感染细胞的数量所测量的。我们的研究还表明，OTA诱导PK-15细胞中的自噬，如通过微管相关蛋白1轻链3（LC 3）-II、自噬相关蛋白5（ATG 5）和Beclin-1的表达显著增加以及绿色荧光蛋白（GFP）-LC 3点的积累所评估的。OTA诱导完全的自噬通量，这是通过监测p62降解和LC 3-II周转使用免疫印迹检测。3-甲基胺（3-MA）和氯喹（CQ）抑制自噬显著减弱OTA诱导的PCV 2复制促进。通过特异性siRNA敲低ATG 5或Beclin-1进一步证实了观察到的现象。进一步的研究表明，活性氧清除剂N-乙酰-L-半胱氨酸（NAC）可以阻断OTA诱导的自噬，表明活性氧可能参与了OTA诱导的自噬的调控。此外，我们观察到与对照组相比，在体内饲喂75和150 μg/kg OTA的猪的肺、脾、肾、肝和腹股沟淋巴结（ILN）和支气管淋巴结（BLN）中OTA浓度显著增加。施用75 μg/kg OTA显著增加猪肺、脾、肾和BLN中的PCV 2复制和自噬。综上所述，可以得出结论，OTA诱导的体外和体内自噬促进PCV 2复制。
Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium. Porcine circovirus type 2 (PCV2) is recognized as the causative agent of porcine circovirus-associated diseases. Recently, we reported that low doses of OTA promoted PCV2 replication in vitro and in vivo, but the underlying mechanism needed further investigation. The present studies further confirmed OTA-induced PCV2 replication promotion as measured by cap protein expression, viral titer, viral DNA copies and the number of infected cells. Our studies also showed that OTA induced autophagy in PK-15 cells, as assessed by the markedly increased expression of microtubule-associated protein 1 light chain 3 (LC3)-II, autophagy-related protein 5 (ATG5), and Beclin-1 and the accumulation of green fluorescent protein (GFP)-LC3 dots. OTA induced complete autophagic flux, which was detected by monitoring p62 degradation and LC3-II turnover using immunoblotting. Inhibition of autophagy by 3-methylademine (3-MA) and chloroquine (CQ) significantly attenuated OTA-induced PCV2 replication promotion. The observed phenomenon was further confirmed by the knock-down of ATG5 or Beclin-1 by specific siRNA. Further studies showed that N-acetyl-L-cysteine (NAC), an ROS scavenger could block autophagy induced by OTA, indicating that ROS may be involved in the regulation of OTA-induced autophagy. Furthermore, we observed significant increases in OTA concentrations in lung, spleen, kidney, liver and inguinal lymph nodes (ILN) and bronchial lymph nodes (BLN) of pigs fed 75 and 150 μg/kg OTA compared with controls in vivo. Administration of 75 μg/kg OTA significantly increased PCV2 replication and autophagy in the lung, spleen, kidney and BLN of pigs. Taken together, it could be concluded that OTA-induced autophagy in vitro and in vivo promotes PCV2 replication.
登录
查看更多内容
DOI：
10.1083/jcb.152.4.657
发表时间：
2001-02-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Mizushima N;Yamamoto A;Hatano M;Kobayashi Y;Kabeya Y;Suzuki K;Tokuhisa T;Ohsumi Y;Yoshimori T
通讯作者：
Yoshimori T
影响因子：
16
作者：
Kroemer G;Mariño G;Levine B
通讯作者：
Levine B
影响因子：
13.3
作者：
Lin WJ;Kuang HY
通讯作者：
Kuang HY
影响因子：
30.3
作者：
Heaton NS;Randall G
通讯作者：
Randall G
自噬维持猪生殖和呼吸道病毒在宿主细胞中的复制
影响因子：
3.7
作者：
Liu Q;Qin Y;Zhou L;Kou Q;Guo X;Ge X;Yang H;Hu H
通讯作者：
Hu H