Effect of Jinzhen granule on two coronaviruses: The novel SARS-CoV-2 and the HCoV-229E and the evidences for their mechanisms of action

Autor: Qinhai, Ma, Zhoulang, Wang, Ruihan, Chen, Biao, Lei, Bin, Liu, Haiming, Jiang, Zexing, Chen, Xuejun, Cai, Xiaowen, Guo, Meihua, Zhou, Jicheng, Huang, Xiaobo, Li, Jun, Dai, Zifeng, Yang
Rok vydání: 2022
Předmět:
CCCP
carbonyl cyanide m-chlorophenylhydrazone
VGM
virus growth medium
Coronaviruses
G-CSF
Granulocyte colony-stimulating factor
Pharmaceutical Science
NHP
nonhuman primates animal model
Mice
MIP-1a
Recombinant Macrophage Inflammatory Protein 1 alpha
Coronavirus 229E
Human
cytC
cytochrome c
Drug Discovery
IP-10
Interferon-inducible protein-10
TNF-α
Tumor necrosis factor-α
MOI
multiplicity of infection
COVID-19
coronavirus disease 2019
IC50
50% inhibition concentrations
DMEM
Dulbecco's modified Eagle's medium
NF-kappa B
virus diseases
HCoV-229E
MERS-CoV
Middle East respiratory syndrome coronavirus
TCID50
50% tissue culture infective dose
PBS
Phosphate-buffered saline
EC50
half maximal effective concentration
Molecular Medicine
HCoV-229E
human coronavirus 229E
TCM
Traditional Chinese medicine
Jinzhen granule
PARP
poly (ADP-ribose) polymerase
RT-qPCR
Real-time Fluorescent Quantitative Polymerase Chain Reaction
MAP Kinase Signaling System
SARS-CoV
severe acute respiratory syndrome coronavirus
SI
selective index
PFU
plaque forming-unit
Antiviral Agents
Article
CPE
Cytopathic effect
FBS
fetal bovine serum
Animals
Humans
JZ
Jinzhen granule
Antiviral
IL-1β
Interleukin-1beta
IL-6
Interleukin-6
GAPDH
Glyceraldehyde 3-phosphate dehydrogenase
Pharmacology
Vero E6
African green monkey kidney epithelial cell line
NF-κB
nuclear transcription factor-kappa B
SARS-CoV-2
COVID-19
DMSO
Dimethyl sulfoxide
MCP-1
Monocyte chemoattracctant protein-1
CC50
50% cytotoxicity concentration
Complementary and alternative medicine
Huh-7
hepatocellular carcinoma cell line
Anti-inflammatory
Drugs
Chinese Herbal
MAPK
mitogen activated protein kinase
Zdroj: Phytomedicine
ISSN: 0944-7113
DOI: 10.1016/j.phymed.2021.153874
Popis: Background Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human coronavirus 229E (HCoV-229E) pose a huge threat to human public health, no specific treatment is available. Jinzhen granule (JZ) is a traditional eight ingredients-Chinese medicine with prominent efficacy for treating viral-induced diseases. However, little is known about the antiviral effect and mechanism of JZ against SARS-CoV-2 and HCoV-229E. Purpose This study aimed to reveal the antiviral effects of JZ against SARS-CoV-2 and HCoV-229E, and to further explore the underlying mechanisms regulating the host immune response. Methods The chromatographic separation of JZ was performed using a Shimadzu analytical high-performance liquid chromatograph with UV detection and Alltech ELSD 2000ES. We conducted cytopathic effect (CPE) and plaque reduction assays to evaluate the antiviral effect of JZ. A lethal human angiotensin converting enzyme 2 (hACE2) transgenic mouse model of SARS-CoV-2 was established to determine the protective effect of JZ on mortality and lung virus titers. Real-time quantitative PCR assays were used to analyze the expression of proinflammatory cytokines in vitro and in vivo. Western blotting was further performed to determine the activities on regulating the nuclear factor kappa B (NF-κB)/MAPK pathway. Finally, mitochondrial membrane potential assays, flow cytometry analysis and western blotting were used to assess the anti-apoptotic potency toward HCoV-229E infection. Results The results showed that 13 chemical components were identified and five peaks were determined and quantitated (gallic acid 1.97 mg/g, baicalin 20.69 mg/g, glycyrrhizic acid 4.92 mg/g, hyodeoxycholic acid 4.86 mg/g, cholic acid 4.07 mg/g). We found that JZ exerted inhibitory potency against SARS-CoV-2 and HCoV-229E in vitro by using CPE and plaque reduction assays, and it was further found that JZ protected mice infected by SARS-CoV-2 from death and inhibited lung virus titers. JZ also significantly decreased the induction of inflammatory cytokines (IL-1α, IL-6, CCL-5 and MIP-1β), similar to the observed in vitro effect. Moreover, JZ suppressed the release of inflammatory cytokines in vitro and it decreased the protein expression of p-p38 MAPK, p-JNK, p-NF-κB p65 and p-IκBα induced by HCoV-229E and increased the expression of IκBα. Notably, JZ significantly protected HCoV-229E-infected Huh-7 cells from mitochondrial damage and decreased apoptotic cells. The activation of the mitochondria-mediated apoptotic pathway was inhibited by JZ, as shown by the reduced expression of cleaved caspase-9, caspase-3 and p-PARP. Conclusions In conclusion, JZ (gallic acid 1.97 mg/g, baicalin 20.69 mg/g, glycyrrhizic acid 4.92 mg/g, hyodeoxycholic acid 4.86 mg/g, cholic acid 4.07 mg/g) exhibited antiviral activities against SARS-CoV-2 and HCoV-229E by regulating the NF-κB/MAPK pathway and the mitochondria-mediated apoptotic pathway. These findings demonstrated the efficacy of JZ against CoVs and suggested JZ treatment as a novel clinical therapeutic strategy for COVID-19.
Databáze: OpenAIRE
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