白藜芦醇抑制猪传染性脑脊髓炎病毒复制。

Suppression of porcine hemagglutinating encephalomyelitis virus replication by resveratrol.

机构信息

Key Laboratory of Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun, China.

Jilin Academy of Animal Husbandry and Veterinary Medicine, Changchun, Jilin, China.

出版信息

Virol J. 2022 Dec 28;19(1):226. doi: 10.1186/s12985-022-01953-5.

DOI:10.1186/s12985-022-01953-5

PMID:36578037

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9795454/

Abstract

BACKGROUND

Porcine hemagglutinating encephalomyelitis virus (PHEV), a member of the genus Betacoronavirus, is the causative agent of neurological disease in pigs. No effective therapeutics are currently available for PHEV infection. Resveratrol has been shown to exert neuroprotective and antiviral effects. Here resveratrol was investigated for its ability to inhibit PHEV replication in nerve cells and central nervous system tissues.

METHODS

Anti-PHEV effect of resveratrol was evaluated using an in vitro cell-based PHEV infection model and employing a mouse PHEV infection model. The collected cells or tissues were used for quantitative PCR analysis, western blot analysis, or indirect immunofluorescence assay. The supernatants were collected to quantify viral loads by TCID assay in vitro. EC50 and CC50 were determined by dose-response experiments, and the ratio (EC50/CC50) was used as a selectivity index (SI) to measure the antiviral versus cytotoxic activity.

RESULTS

Our results showed that resveratrol treatment reduced PHEV titer in a dose-dependent manner, with a 50% inhibition concentration of 6.24 μM. A reduction of > 70% of viral protein expression and mRNA copy number and a 19-fold reduction of virus titer were achieved when infected cells were treated with 10 µM resveratrol in a pre-treatment assay. Quantitative PCR analysis and TCID assay results revealed that the addition of 10 μM resveratrol to cells after adsorption of PHEV significantly reduced 56% PHEV mRNA copy number and eightfold virus titer. 10 µM resveratrol treatment reduced 46% PHEV mRNA copy number and fourfold virus titer in virus inactivation assay. Moreover, the in vivo data obtained in this work also demonstrated that resveratrol inhibited PHEV replication, and anti-PHEV activities of resveratrol treatment via intranasal installation displayed better than oral gavage.

CONCLUSION

These results indicated that resveratrol exerted antiviral effects under various drug treatment and virus infection conditions in vitro and holds promise as a treatment for PHEV infection in vivo.

摘要

背景

猪传染性脑脊髓炎病毒（PHEV）是冠状病毒属的一个成员，是引起猪神经疾病的病原体。目前尚无有效的治疗方法可用于 PHEV 感染。白藜芦醇已被证明具有神经保护和抗病毒作用。本研究旨在探讨白藜芦醇抑制神经细胞和中枢神经系统组织中 PHEV 复制的能力。

方法

采用体外细胞 PHEV 感染模型和小鼠 PHEV 感染模型评价白藜芦醇的抗 PHEV 作用。收集细胞或组织进行定量 PCR 分析、western blot 分析或间接免疫荧光检测。收集上清液通过 TCID 测定法在体外定量病毒载量。通过剂量反应实验确定 EC50 和 CC50，并使用 EC50/CC50 比值作为选择性指数（SI）来衡量抗病毒与细胞毒性活性。

结果

结果显示，白藜芦醇处理呈剂量依赖性降低 PHEV 滴度，50%抑制浓度为 6.24 μM。在预处理试验中，感染细胞用 10 μM 白藜芦醇处理时，病毒蛋白表达和 mRNA 拷贝数减少>70%，病毒滴度降低 19 倍。定量 PCR 分析和 TCID 测定结果显示，在吸附 PHEV 后向细胞中加入 10 μM 白藜芦醇可显著降低 56%的 PHEV mRNA 拷贝数和 8 倍的病毒滴度。病毒失活试验中，10 μM 白藜芦醇处理可降低 46%的 PHEV mRNA 拷贝数和 4 倍的病毒滴度。此外，本工作的体内数据还表明，白藜芦醇抑制 PHEV 复制，经鼻腔给药的白藜芦醇抗 PHEV 活性优于口服灌胃。