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Objective:To studytheinhibitoryeffectsof antisenseoligodeoxynucleotide(ASODN)anditsthiophosphate(S-ASODN)on thereplicationandexpressionof hepatitisD virus(HDV)in H1δ9cellsandin tupaiabody.Methods:After15mer-ASODNandS-ASODNweresynthesized,differentconcentrationsof ASODNandS-ASODNwereaddedto theculturemediumof H1δ9celllineandthenHDAgin thesupernatantof theculturewas examinedwithELISAand HDV-RNAinthecellsdeterminedwithdotblothybridization.SixteentupaiaeweresuccessfullyinfectedwithHDVand thenequallyrandomizedinto2groups.In thetreatedgroup,theanimalsreceivedintravenousinjectionsof S-ASODN3mg everyotherdayfor7timesandthecontrolgroupwereinjected withsamevolumeof normalsaline.On the5th,10th,15thand20thdayaftertheadministration,samplesof bloodandlivertissueswereexaminedwithimmunohistochemical methodforHDAganddotblothybridizationandin situ hybridizationforHDV-RNA.Results:Twenty-fourhoursafter theadditionof a finalconcentrationof6μmol/Lof S-ASODN,thereplicationof HDV-RNAandthereleaseof HDAgin theH1δ9cellsweresuppressedby84.5%and76.14%respectively.Theinhibitionwas dose-dependentwhenthefinal concentrationsof2,4and6μmol/Lof S-ASODNweregiven.WhenASODNandS-ASODNwereadministeredinthe samedosage,theinhibitionshowedno significantlydifferencebetweenthe2agents.On thelastdayof theadministration of S-ASODN,7outof the8tupaiaeof thetreatedgroupshowednegativeHDAgandHDV-RNAinthelivertissuewhile only1outof the8tupaiaeof thecontrolgroupwasnegative.Tendaysafterthecessationof drugadministration,3tupaiae of thetreatedgroupand7of controlwerepositiveof HDAgandHDV-RNA.Conclusion:OurfindingsshowthatS-ASODNefficientlyinhibitesthereplicationandexpressionof HDVgeneinH1δ9cellsandin thebodyof tupaia,which providesanexperimentalbasisfortheanti-HDVapplicationof antisenseoligonucleotides.
Objective: To studytheinhibitoryeffectsof antisenseoligodeoxynucleotide (ASODN) anditsthiophosphate (S-ASODN) on thereplicationandexpressionof hepatitisD virus (HDV) in H1δ9cellsandin tupaiabody.Methods: After15mer-ASODNandS-ASODNweresynthesized, differentconcentrationsof ASODNandS-ASODNwereaddedto theculturemediumof H1δ9celllineandthenHDAgin thesupernatantof theculturewas examinedwithELISAand HDV-RNAinthecellsdeterminedwithdotblothybridization.SixteentupaiaeweresuccessfullyinfectedwithHDVand thenequallyrandomizedinto2groups.In thetreatedgroup, theanimalsreceivedintravenousinjectionsof S-ASODN3mg everyotherdayfor7timesandthecontrolgroupwereinjected withsamevolumeof normalsaline.On the5th, 10th, 15thand20thdayaftertheadministration, samplesof bloodandlivertissueswereexaminedwithimmunohistochemical methodforHDAganddotblothybridizationandin situ hybridizationforHDV-RNA.Results: Twenty-fourhoursafter theadditionof a finalconcentrationof6μmol / Lof S-ASODN, thereplicationof HDV-RNAandthereleaseof HDAgin theH1δ9cellsweresuppressedby84.5% and76.14% respectively.Theinhibitionwas dose-dependentwhenthefinal concentrationsof2,4and6μmol / Lof S-ASODNweregiven.WhenASODNandS-ASODNwereadministeredinthe samedosage, theinhibitionshowedno significantlydifferencebetweenthe2agents.On thelastdayof theadministration of S-ASODN, 7outof the8tupaiaeof thetreatedgroupshowednegativeHDAgandHDV-RNAinthelivertissuewhile only1outof the8tupaiaeof thecontrolgroupwasnegative.Tendaysafterthecessationof drugadministration , 3tupaiae of thetreated group and 7of controlwerepositiveof HDAgandHDV-RNA.Conclusion: OurfindingsshowthatS-ASODNefficientlyhibitsthereplicationandexpressionofHVVgeneinH1δ9cellsandin thebodyof tupaia, which providesanexperimentalbasisfortheanti-HDVapplicationof antisenseoligonucleotides.