Dermcidin促进猪流行性腹泻病毒复制的研究

Dermcidin promotes replication of porcine epidemic diarrhea virus

猪流行性腹泻病(PED)是由猪流行性腹泻病毒(PEDV)引起的一种高度传染性的猪胃肠道疾病,可导致哺乳仔猪产生急性水样腹泻,造成严重的经济损失。为调查抗菌肽Dermcidin(DCD)对PEDV复制的影响,本研究利用Western blot、荧光定量PCR(RT-qPCR)以及测定病毒半数组织细胞感染量TCID_(50)等方法,通过检测在DCD的作用下PEDV在细胞中的基因转录、蛋白表达和病毒滴度的水平,并利用Western blot检测信号通路中蛋白表达水平,探究其作用的分子机制。研究表明,DCD能够参与PEDV感染;DCD过表达促进了PEDV在Vero细胞和LLC-PK1细胞中的复制,而敲低DCD抑制了PEDV复制;DCD通过诱导PTEN表达来抑制Akt-mTOR-p70S6K通路相关蛋白位点的磷酸化,从而介导自噬促进PEDV复制。研究DCD对PEDV复制的作用机制,对今后PEDV防治工作有重要参考意义。

Porcine epidemic diarrhoea(PED) is a highly infectious porcine gastrointestinal disease caused by porcine epidemic diarrhoea virus(PEDV), which can lead to acute watery diarrhoea in lactating piglets and cause severe economic losses. To investigate the effect of the antimicrobial peptide Dermcidin(DCD) on PEDV replication, this study used Western blot, fluorescent quantitative PCR(RT-qPCR) and the determination of the viral half tissue cell infection TCID_(50)to detect the levels of PEDV gene transcription, protein expression and viral titer in cells in the presence of DCD, and The molecular mechanism of its action was investigated by detecting protein expression levels in the signaling pathway using Western blot. The study showed that DCD was involved in PEDV infection;DCD overexpression promoted PEDV replication in Vero and LLC-PK1 cells, while knockdown of DCD inhibited PEDV replication;DCD inhibited phosphorylation of Akt-mTOR-p70S6K pathway-associated protein sites by inducing PTEN expression, thereby mediating autophagy to promote PEDV replication. The study of the mechanism of action of DCD on PEDV replication will be an important reference for future PEDV preve ntion and treatment.