背景:葡萄糖调节蛋白94(Grp94)是病毒后生命周期所必需的,并且在病毒蛋白中起质量控制作用,但是Grp94在宿主细胞中调节病毒复制的作用尚不清楚。因此,找到一种能调控Grp94的化合物将有助于我们研究病毒复制的机制。以前,我们合成了香豆素吡唑啉衍生物HCP1,它是一种有效的Grp94抑制剂。我们推测HCP1可能抑制病毒复制。
目的:本研究旨在研究HCP1对SenecavirusA(SVA)复制能力的影响,从而为揭示病毒的致病机理和开发抗病毒药物提供靶标和先导化合物。
方法:用SVA感染大鼠细胞系BHK-21和猪细胞系PK-15,用不同浓度的HCP1处理感染的细胞。细胞活力(CCK-8),病毒滴度(TCID50),自噬水平,测量Grp94表达。
结果:结果表明,低浓度的HCP1降低了BHK-21和PK-15细胞中的病毒滴度和病毒载量,5μMHCP1显著降低了SVAVP2蛋白的表达。此外,SVA感染可导致自噬水平升高,HCP1能抑制SVA感染引起的宿主细胞自噬,从而抑制病毒复制和感染。
结论:这些发现表明,Grp94是控制SVA复制的关键因素,其抑制剂HCP1通过抑制Grp94蛋白水平的升高和SVA诱导的自噬来抑制SVA的复制。这项研究将有助于开发新一类小分子抗病毒药物。
BACKGROUND: Glucoregulatory protein 94 (
Grp94) is necessary for the post-viral life cycle and plays a quality control role in viral proteins, but the role of
Grp94 in regulating viral replication in host cells is not well known. Therefore, finding a compound that can regulate Grp94 will help us to study the mechanism of viral replication. Previously, we synthesized a coumarin pyrazoline derivative HCP1 that is an effective inhibitor of
Grp94. We suppose that HCP1 may inhibit viral replication.
OBJECTIVE: This study aimed to investigate the effect of HCP1 on the replication ability of Senecavirus A (SVA), so as to provide a target and a leading compound for revealing the pathogenic mechanism of the virus and developing antiviral drugs.
METHODS: Rat cell lines BHK-21 and porcine cell lines PK-15 were infected with SVA, and the infected cells were treated with different concentrations of HCP1. The cell viability (CCK-8), virus titer (TCID50), autophagy level, and
Grp94 expression were measured.
RESULTS: The results showed that a low concentration of HCP1 decreased viral titer and viral load in BHK-21 and PK-15 cells, and 5μM HCP1 significantly decreased the expression of SVA VP2 protein. In addition, SVA infection can lead to an increased level of autophagy, and HCP1 can inhibit host cell autophagy caused by SVA infection, thereby inhibiting viral replication and infection.
CONCLUSIONS: These findings reveal that Grp94 is a key factor in controlling SVA replication, and its inhibitor HCP1 suppresses SVA replication by inhibiting the increase of
Grp94 protein level and autophagy induced by SVA. This study will contribute to the development of a new class of small-molecule antiviral drugs.