OBJECTIVE: In vitro and in vivo PEDV infection models were used to evaluate the anti-PEDV potential of LZD extract.
METHODS: LC-MS was used for qualitative analysis of LZD. The antiviral effect of LZD against PEDV using flow cytometry (FC), Quantitative real-time polymerase chain reaction (QPCR), immunofluorescence assay (IFA) analysis in Vero and IPEC-J2 cells. Additionally, we measured the survival rate, clinical symptoms, body weights, fecal scores, temperature, histological analysis, and viral load in a model of newborn piglets infected with PEDV in order to assess the antiviral impact of LZD in vivo.
RESULTS: In total, 648 compounds were identified, including 144 Alkaloids, 128 Terpenoids, etc. LZD effectively suppressed PEDV replication in vitro. According to time of addition experiments, LZD mostly inhibited PEDV during the viral life cycle\'s replication stages. During PEDV infection, LZD can Significantly decrease the apoptotic rate of IPEC-J2 cells and Vero cells. In comparison to the model group, LZD was able to decrease the viral titers in the infected piglets\' intestinal and visceral tissues, ameliorate their intestinal pathology, cause a significant increase in body weight growth and increase the piglet survival rate.
CONCLUSIONS: Our findings indicate that the aqueous solution derived from LZD suppressed PEDV replication both in vitro and in vivo, indicating its potential as a candidate for pharmaceutical development.
目的:使用体外和体内PEDV感染模型来评估LZD提取物的抗PEDV潜力。
方法:LC-MS用于LZD的定性分析。使用流式细胞术(FC)研究LZD对PEDV的抗病毒作用,定量实时聚合酶链反应(RT-qPCR),Vero和IPEC-J2细胞中的免疫荧光测定(IFA)分析。此外,我们测量了存活率,临床症状,体重,粪便评分,温度,组织学分析,和感染PEDV的新生仔猪模型中的病毒载量,以评估LZD在体内的抗病毒作用。
结果:总计,鉴定出648种化合物,包括144种生物碱,128萜类化合物,等。LZD在体外有效抑制PEDV复制。根据添加实验的时间,LZD在病毒生命周期的复制阶段主要抑制PEDV。在PEDV感染期间,LZD可显著降低IPEC-J2细胞和Vero细胞的凋亡率。与模型组相比,LZD能够降低感染仔猪肠道和内脏组织中的病毒滴度,改善他们的肠道病理学,导致体重显著增加,增加仔猪成活率。
结论:我们的发现表明,源自LZD的水溶液在体外和体内均抑制了PEDV的复制,表明其作为药物开发候选人的潜力。