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1、AbstractAssessmentofMitochondrialToxicityinHepG2CellsTreatedwithCepharanthineHydrochloridePostgraduate:YafeiZhangSupervisor:Prof.QingduanWangDepartmentofPharmacology,ZhengzhouUniversity,HenanZhengzhou450052AbsractBackgroundandObjective:CepharanthineHydrochloride,abisbenzyisoquinolinea
2、lkaloid,isextractedfromstephaniacepharanthaHayataandthensalificatiedwithhydrochloricacid.Ithasmanybiologicalandpharmacologicalactivities,includinganti-inflammatory,immuneregulation,thedifferentiationoftumorcells,andincreaseofleukocyteactivity,inductionofapoptosis,painreliefandmultidru
3、gresistancereversal.ThehumanhepatomaHepG2cellsarerichinmitochondrialtoxicityandmitochondrialDNA,soitisthebestcellmodeltostudymitochondrialtoxicity.ThepresentresearchwasconductedtoexplorethepotentialmitochondrialtoxicityofCepharanthineHydrochlorideinHepG2cells,andtoprovidetheclinicalre
4、ferencesforsafeuseofthedrug.Method:WeadoptMTTtoselectarangeoflowerthantheIC50asafurtherscreeningconcentration,anduseHepG2cellsasempiricalmodel.Theexperimenttestsetupfivegroups:CHhigh（0.1μM）、medium（0.05μM）、lowdose（0.025μM）group，zidovudine(AZT)groupasthepositivecontroldrug(10μM),Solvent
5、controlgroup.Drugeffectscellfor9day,wedetectedlactatelevelonHepG2cellsbyLacticIII万方数据AbstractAcid(LD)assaykit;extractedmitochondriainHepG2cellsbyChemicaltreatmentmethod;determinedproteinconcentrationinmitochondriabyStandardTestMethod;assayedtheactivityofmitochondrialrespiratorychainco
6、mplexesII,IVbySpectrophotometricmethodandtheeffectofCHonreplicationofmitochondrialDNAinHepG2cellsbyPolymerasechainreaction(PCR)method,weusethemethodoffluorescencequantitativePCRtodynamicobservationofchangesofmtDNAineachgroup,observethemorphologyanddistributionofmitochondriabyTransmiss
7、ionelectronmicroscopy.Result:1)TheinhibitionrateofcellproliferationbyCHhigh、medium、lowdosegroupandzidovudine(AZT)groupwere(1.63±0.15)%、(1.49±0.13)%、(0.47±0.08)%and（0.90±0.10）%,respectively.2)TheconcentrationoflacticacidinCHhigh、medium、lowdosegroup,zidovudine(AZT)groupsolventcontrolgro
8、upwer
