PMID- 33677370 OWN - NLM STAT- MEDLINE DCOM- 20210920 LR - 20211204 IS - 1873-2542 (Electronic) IS - 0378-1135 (Linking) VI - 255 DP - 2021 Apr TI - Small molecule screening identified cepharanthine as an inhibitor of porcine reproductive and respiratory syndrome virus infection in vitro by suppressing integrins/ILK/RACK1/PKCalpha/NF-kappaB signalling axis. PG - 109016 LID - S0378-1135(21)00039-0 [pii] LID - 10.1016/j.vetmic.2021.109016 [doi] AB - Porcine Reproductive and Respiratory Syndrome (PRRS) is a devastating disease among the most notorious threats to the swine industry worldwide and is characterized by respiratory distress and reproductive failure. Highly evolving porcine reproductive and respiratory syndrome virus (PRRSV) strains with complicated genetic diversity make the current vaccination strategy far from cost-effective and thus urge identification of potent lead candidates to provide prevention and treatment approaches. From an in vitro small molecule screening with the TargetMol Natural Compound Library comprising 623 small molecules, cytopathic effect (CPE) observations and RT-qPCR analysis of viral ORF7 gene expression identified cepharanthine (CEP) to be one of the most protent inhibitors of PRRSV infection in Marc-145 cells. When compared with tilmicosin, which is one of the most commonly used antibiotics in swine industry to inhibit infections, CEP more prominently inhibited PRRSV infection represented by both RNA and protein levels, further reduced the TCID(50) by 5.6 times, and thus more remarkably protected Marc-145 cells against PRRSV infection. Mechanistically, western blot analyses of the Marc-145 cells and the porcine alveolar macrophages (PAMs) with or without CEP treatment and PRRSV infection at various time points revealed that CEP can inhibit the expression of integrins beta1 and beta3, integrin-linked kinase (ILK), RACK1 and PKCalpha, leading to NF-kappaB suppression and consequent alleviation of PRRSV infection. Collectively, our small molecule screening identified cepharanthine as an inhibitor of PRRSV infection in vitro by suppressing Integrins/ILK/RACK1/PKCalpha/NF-kappaB signalling axis, which may enlighten the deeper understanding of the molecular pathogenesis of PRRSV infection and more importantly, suggested CEP as a potential promising drug for PRRS control in veterinary clinics. CI - Copyright (c) 2021 Elsevier B.V. All rights reserved. FAU - Yang, Chao AU - Yang C AD - College of Animal Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, Yunnan, China. FAU - Zuo, Qingwei AU - Zuo Q AD - College of Animal Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, Yunnan, China. FAU - Liu, Xiao AU - Liu X AD - College of Animal Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, Yunnan, China; Department of Oncology-Pathology, Karolinska Institute, 17176, Stockholm, Sweden. FAU - Zhao, Qian AU - Zhao Q AD - Center for Animal Disease Control and Prevention, Chuxiong, 675000, Yunnan, China. FAU - Pu, Haoyu AU - Pu H AD - College of Animal Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, Yunnan, China. FAU - Gao, Libo AU - Gao L AD - College of Animal Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, Yunnan, China. FAU - Zhao, Lianfeng AU - Zhao L AD - College of Life Sciences, Nanjing Normal University, Wenyuan Rd. No. 1, Nanjing, 210023, Jiangsu, China. FAU - Guo, Zhigang AU - Guo Z AD - College of Life Sciences, Nanjing Normal University, Wenyuan Rd. No. 1, Nanjing, 210023, Jiangsu, China. FAU - Lin, Yingbo AU - Lin Y AD - Department of Oncology-Pathology, Karolinska Institute, 17176, Stockholm, Sweden. FAU - Liu, Jianping AU - Liu J AD - College of Life Sciences, Nanjing Normal University, Wenyuan Rd. No. 1, Nanjing, 210023, Jiangsu, China. FAU - Bi, Junlong AU - Bi J AD - Institute of Science and Technology, Chuxiong Normal University, 546 Lucheng South Rd, Chuxiong, 675000, Yunnna, China. Electronic address: 419893334@qq.com. FAU - Yin, Gefen AU - Yin G AD - College of Animal Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, Yunnan, China. Electronic address: yingefen0616@163.com. LA - eng PT - Journal Article DEP - 20210217 PL - Netherlands TA - Vet Microbiol JT - Veterinary microbiology JID - 7705469 RN - 0 (Antiviral Agents) RN - 0 (Benzylisoquinolines) RN - 0 (Integrins) RN - 0 (NF-kappa B) RN - 0 (Receptors for Activated C Kinase) RN - 7592YJ0J6T (cepharanthine) RN - EC 2.7.1.- (integrin-linked kinase) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.13 (Protein Kinase C-alpha) SB - IM MH - Animals MH - Antiviral Agents/pharmacology MH - Benzylisoquinolines/*pharmacology MH - Cell Line MH - Chlorocebus aethiops MH - Gene Expression Regulation/drug effects MH - Integrins/genetics/*metabolism MH - NF-kappa B/genetics/metabolism MH - Porcine Reproductive and Respiratory Syndrome/drug therapy/virology MH - *Porcine respiratory and reproductive syndrome virus MH - Protein Kinase C-alpha/genetics/*metabolism MH - Protein Serine-Threonine Kinases/genetics/*metabolism MH - Receptors for Activated C Kinase/genetics/*metabolism MH - Signal Transduction MH - Swine OTO - NOTNLM OT - Cepharanthine OT - Integrin OT - Integrin-linked kinase OT - NF-kappaB OT - PKCalpha OT - Porcine reproductive and respiratory syndrome virus OT - RACK1 EDAT- 2021/03/08 06:00 MHDA- 2021/09/21 06:00 CRDT- 2021/03/07 20:37 PHST- 2020/12/02 00:00 [received] PHST- 2021/02/14 00:00 [accepted] PHST- 2021/03/08 06:00 [pubmed] PHST- 2021/09/21 06:00 [medline] PHST- 2021/03/07 20:37 [entrez] AID - S0378-1135(21)00039-0 [pii] AID - 10.1016/j.vetmic.2021.109016 [doi] PST - ppublish SO - Vet Microbiol. 2021 Apr;255:109016. doi: 10.1016/j.vetmic.2021.109016. Epub 2021 Feb 17.