Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained wi...Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained with OilRed-O staining to confirm adipogenesis.The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay,intracellular reactive oxygen species generation through flow cytometry,and morphologically through confocal microscopy using propidium iodide,4,6-diamino-2-phenylindol dihydrochloride,and 2’,7’-dichlorofluorescein diacetate dyes.The uptake of fluorescent glucose analog,2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy.Results:MTT assay showed that the cell survival rate was(28.00±3.00)%,(92.33±2.60)%,and(71.34±2.10)%in the presence of 2 mM H2O2,AbPI alone,and AbPI and H2O2 both,respectively,in comparison to the control.Oil-Red-O staining indicated that Ab PI enhanced adipogenesis.AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone,and showed insulinsensitizing effect in the presence of insulin,but failed to stimulate the uptake in the absence of insulin.Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon Ab PI treatment.Confocal microscopy showed that the damaged cell population rose to 3.50%,117.84%,and 261.50%in the presence of Ab PI alone,AbPI with H2O2,and H2O2 alone,respectively.Conclusions:The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.展开更多
Unlike human immunodeficiency virus(HIV)and hepatitis B virus(HBV),hepatitis C virus(HCV)infection is a curable disease.Current direct antiviral agent(DAA)targets are focused on HCV NS3/4A protein(protease),NS5 B prot...Unlike human immunodeficiency virus(HIV)and hepatitis B virus(HBV),hepatitis C virus(HCV)infection is a curable disease.Current direct antiviral agent(DAA)targets are focused on HCV NS3/4A protein(protease),NS5 B protein(polymerase)and NS5 A protein.The first generation of DAAs includes boceprevir and telaprevir,which are protease inhibitors and were approved for clinical use in2011.The cure rate for genotype 1 patients increased from 45% to 70% when boceprevir or telaprevir was added to standard PEG-IFN/ribavirin.More effective and less toxic second generation DAAs supplanted these drugs by 2013.The second generation of DAAs includes sofosbuvir(Sovaldi),simeprevir(Olysio),and fixed combination medicines Harvoni and Viekira Pak.These drugs increase cure rates to over 90%without the need for interferon and effectively treat all HCV genotypes.With these drugs the "cure HCV"goal has become a reality.Concerns remain about drug resistance mutations and the high cost of these drugs.The investigation of new HCV drugs is progressing rapidly;fixed dose combination medicines in phase III clinical trials include Viekirax,asunaprevirtdaclatasvirtbeclabuvir,grazoprevirtelbasvir and others.展开更多
文摘Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained with OilRed-O staining to confirm adipogenesis.The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay,intracellular reactive oxygen species generation through flow cytometry,and morphologically through confocal microscopy using propidium iodide,4,6-diamino-2-phenylindol dihydrochloride,and 2’,7’-dichlorofluorescein diacetate dyes.The uptake of fluorescent glucose analog,2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy.Results:MTT assay showed that the cell survival rate was(28.00±3.00)%,(92.33±2.60)%,and(71.34±2.10)%in the presence of 2 mM H2O2,AbPI alone,and AbPI and H2O2 both,respectively,in comparison to the control.Oil-Red-O staining indicated that Ab PI enhanced adipogenesis.AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone,and showed insulinsensitizing effect in the presence of insulin,but failed to stimulate the uptake in the absence of insulin.Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon Ab PI treatment.Confocal microscopy showed that the damaged cell population rose to 3.50%,117.84%,and 261.50%in the presence of Ab PI alone,AbPI with H2O2,and H2O2 alone,respectively.Conclusions:The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.
文摘Unlike human immunodeficiency virus(HIV)and hepatitis B virus(HBV),hepatitis C virus(HCV)infection is a curable disease.Current direct antiviral agent(DAA)targets are focused on HCV NS3/4A protein(protease),NS5 B protein(polymerase)and NS5 A protein.The first generation of DAAs includes boceprevir and telaprevir,which are protease inhibitors and were approved for clinical use in2011.The cure rate for genotype 1 patients increased from 45% to 70% when boceprevir or telaprevir was added to standard PEG-IFN/ribavirin.More effective and less toxic second generation DAAs supplanted these drugs by 2013.The second generation of DAAs includes sofosbuvir(Sovaldi),simeprevir(Olysio),and fixed combination medicines Harvoni and Viekira Pak.These drugs increase cure rates to over 90%without the need for interferon and effectively treat all HCV genotypes.With these drugs the "cure HCV"goal has become a reality.Concerns remain about drug resistance mutations and the high cost of these drugs.The investigation of new HCV drugs is progressing rapidly;fixed dose combination medicines in phase III clinical trials include Viekirax,asunaprevirtdaclatasvirtbeclabuvir,grazoprevirtelbasvir and others.
