RESUMO
In this study, we demonstrate the synthesis of carbonized nanogels (CNGs) from an amino acid (lysine hydrochloride) using a simple pyrolysis method, resulting in effective viral inhibition properties against infectious bronchitis virus (IBV). The viral inhibition of CNGs was studied using both in vitro (bovine ephemeral fever virus (BEFV) and pseudorabies virus (PRV)) and in ovo (IBV) models, which indicated that the CNGs were able to prevent virus attachment on the cell membrane and penetration into the cell. A very low concentration of 30 µg mL-1 was found to be effective (>98% inhibition) in IBV-infected chicken embryos. The hatching rate and pathology of IBV-infected chicken embryos were greatly improved in the presence of CNGs. CNGs with distinctive virus-neutralizing activities show great potential as a virostatic agent to prevent the spread of avian viruses and to alleviate the pathology of infected avian species.
Assuntos
Antivirais/farmacologia , Infecções por Coronavirus/tratamento farmacológico , Vírus da Bronquite Infecciosa/efeitos dos fármacos , Lisina/farmacologia , Nanogéis/administração & dosagem , Substâncias Protetoras/farmacologia , Animais , Linhagem Celular , Galinhas/virologia , Chlorocebus aethiops , Infecções por Coronavirus/virologia , Cricetinae , Vírus da Febre Efêmera Bovina/efeitos dos fármacos , Feminino , Herpesvirus Suídeo 1/efeitos dos fármacos , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/virologia , Ratos , Ratos Sprague-Dawley , Células Vero , Internalização do Vírus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
The non-coding microRNA (miRNA) is involved in the regulation of hepatitis C virus (HCV) infection and offers an alternative target for developing anti-HCV agent. In this study, we aim to identify novel cellular miRNAs that directly target the HCV genome with anti-HCV therapeutic potential. Bioinformatic analyses were performed to unveil liver-abundant miRNAs with predicted target sequences on HCV genome. Various cell-based systems confirmed that let-7b plays a negative role in HCV expression. In particular, let-7b suppressed HCV replicon activity and down-regulated HCV accumulation leading to reduced infectivity of HCVcc. Mutational analysis identified let-7b binding sites at the coding sequences of NS5B and 5'-UTR of HCV genome that were conserved among various HCV genotypes. We further demonstrated that the underlying mechanism for let-7b-mediated suppression of HCV RNA accumulation was not dependent on inhibition of HCV translation. Let-7b and IFNα-2a also elicited a synergistic inhibitory effect on HCV infection. Together, let-7b represents a novel cellular miRNA that targets the HCV genome and elicits anti-HCV activity. This study thereby sheds new insight into understanding the role of host miRNAs in HCV pathogenesis and to developing a potential anti-HCV therapeutic strategy.
Assuntos
Hepacivirus/fisiologia , MicroRNAs/genética , Regiões 5' não Traduzidas , Antivirais/farmacologia , Sequência de Bases , Linhagem Celular , Biologia Computacional , Primers do DNA/genética , Genoma Viral , Hepacivirus/efeitos dos fármacos , Hepacivirus/genética , Humanos , Interferon-alfa/farmacologia , Fígado/metabolismo , Fígado/virologia , MicroRNAs/metabolismo , Mutagênese , Polietilenoglicóis/farmacologia , RNA Viral/genética , RNA Viral/metabolismo , Proteínas Recombinantes/farmacologia , Replicon , Proteínas não Estruturais Virais/genética , Replicação Viral/efeitos dos fármacos , Replicação Viral/genéticaRESUMO
The filamentous fungus Monascus pilosus was genetically transformed with a reporter plasmid, pMS-1.5hp, by aurintricarboxylic acid (ATA) treatment to obtain an efficient red-pigment producing mutant. The transformation efficiency of Monascus pilosus was higher with the ATA-treatment than with either a non-restriction-enzyme-mediated integration (REMI) or a REMI method. This valid and convenient random mutagenesis method shows that ATA can be applied in fungi for efficient genetic transformation.