Background Japanese encephalitis virus(JEV)is a leading cause of viral encephalitis worldwide.JEV exhibits significant neuroinvasiveness and neurotoxicity,resulting in considerable damage to the nervous system.Japanes...Background Japanese encephalitis virus(JEV)is a leading cause of viral encephalitis worldwide.JEV exhibits significant neuroinvasiveness and neurotoxicity,resulting in considerable damage to the nervous system.Japanese encephalitis is associated with high morbidity and mortality rate,seriously harming both human health and livestock production.The current lack of specific antiviral drugs means that the development of new therapeutic agents for JEV has become urgent.Methods Anti-JEV drugs were screened from 111 inhibitors of neurotransmitter receptor-related molecules by high content technology.The antiviral effects of clomipramine HCl were evaluated through plaque assay,real-time quantitative PCR,immunofluorescence assay and western blotting assay.Bioinformatic tools were utilized to cluster the altered signaling pathway members after clomipramine HCl treatment.Finally,the anti-JEV mechanism was deeply resolved in vivo via such molecular biology and virological detection techniques.Results In this study,we screened nine compounds with significant anti-JEV activity,of which clomipramine HCl demonstrated the most potent antiviral effect and exhibited dose-dependent activity.Mechanistically,clomipramine HCl may activate endoplasmic reticulum stress and modulate the unfolded protein response,thus inhibiting the assembly stage of JEV infection.Conclusion This study highlights the importance of clomipramine HCl as a promising approach for JEV infection protection,which may lead to new host-directed antiviral approaches to such mosquito-borne viruses.展开更多
High-throughput measurements of ciprofloxacin, clomipramine and fexofenadine hydrochlorides were performed by employing an automatic 8-channel electrical titrator. Silver nitrate （AgNO3） and sodium tetraphenylborate...High-throughput measurements of ciprofloxacin, clomipramine and fexofenadine hydrochlorides were performed by employing an automatic 8-channel electrical titrator. Silver nitrate （AgNO3） and sodium tetraphenylborate （NaTPB） were used as titrants. When AgNO3 was used for measuring the drugs in pure form, recoveries were 97.6%-102.0% with RSD values ≤1.0%; for measuring them in pharmaceutical formulations, recoveries were 96.6%-99.1% with RSD values ≤1.0%. Batch samples of eight could be measured simultaneously and maximally 30 measurements per minute could be completed. When NaTPB was used for measuring the drugs in pure form, the recoveries were 96.8%-102.6% with RSD values 〈0.8%; for measuring them in pharmaceutical formulations, the recoveries were 97.5%-102.7% with RSD values ≤0.9%. For all analyses, no auxiliary devices or chemicals were needed and there was no requirement for changing or cleaning working electrodes between measurements. The efficiency, accuracy and precision of the proposed method make it an alternative for routine quality control analyses.展开更多
基金National Key Research and Development Program of China(2022YFD1801500,2022YFD1800105,2021YFC2600204)National Natural Science Foundation of China(32022082,31825025,32030107).
文摘Background Japanese encephalitis virus(JEV)is a leading cause of viral encephalitis worldwide.JEV exhibits significant neuroinvasiveness and neurotoxicity,resulting in considerable damage to the nervous system.Japanese encephalitis is associated with high morbidity and mortality rate,seriously harming both human health and livestock production.The current lack of specific antiviral drugs means that the development of new therapeutic agents for JEV has become urgent.Methods Anti-JEV drugs were screened from 111 inhibitors of neurotransmitter receptor-related molecules by high content technology.The antiviral effects of clomipramine HCl were evaluated through plaque assay,real-time quantitative PCR,immunofluorescence assay and western blotting assay.Bioinformatic tools were utilized to cluster the altered signaling pathway members after clomipramine HCl treatment.Finally,the anti-JEV mechanism was deeply resolved in vivo via such molecular biology and virological detection techniques.Results In this study,we screened nine compounds with significant anti-JEV activity,of which clomipramine HCl demonstrated the most potent antiviral effect and exhibited dose-dependent activity.Mechanistically,clomipramine HCl may activate endoplasmic reticulum stress and modulate the unfolded protein response,thus inhibiting the assembly stage of JEV infection.Conclusion This study highlights the importance of clomipramine HCl as a promising approach for JEV infection protection,which may lead to new host-directed antiviral approaches to such mosquito-borne viruses.
基金supported by Special Scientific Research Funds for Central Non-profit Institutes,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences(YSFRI-CAFS)(No.20603022016003)Import of International Advanced Agricultural Science and Technology Plan(948 Project)of Chinese Ministry of Agriculture(No.2016-X28)+2 种基金Central Public Interest Scientific Institution Basal Research Fund,CAFS(No.2016RC-BR02)Qingdao National Laboratory for Marine Science and Technology(No.2015ASKJ02-05)Key R&D Program of Shandong Province(No.2016GSF120008)
文摘High-throughput measurements of ciprofloxacin, clomipramine and fexofenadine hydrochlorides were performed by employing an automatic 8-channel electrical titrator. Silver nitrate （AgNO3） and sodium tetraphenylborate （NaTPB） were used as titrants. When AgNO3 was used for measuring the drugs in pure form, recoveries were 97.6%-102.0% with RSD values ≤1.0%; for measuring them in pharmaceutical formulations, recoveries were 96.6%-99.1% with RSD values ≤1.0%. Batch samples of eight could be measured simultaneously and maximally 30 measurements per minute could be completed. When NaTPB was used for measuring the drugs in pure form, the recoveries were 96.8%-102.6% with RSD values 〈0.8%; for measuring them in pharmaceutical formulations, the recoveries were 97.5%-102.7% with RSD values ≤0.9%. For all analyses, no auxiliary devices or chemicals were needed and there was no requirement for changing or cleaning working electrodes between measurements. The efficiency, accuracy and precision of the proposed method make it an alternative for routine quality control analyses.
