| Berberine,extracted from Rhizoma Coptidis, Amur Corktree Bark,Berberis vernaeSchneid and other Chinesis medicine herbs,belong to isoquinoline alkaloids,was aquaternaryammonium compound.The pharmacological properties of berberine includingantibiosis,anti-inflammatory,antiviral,antiarrhythmias,resisting heart failure,protectingischemic cardiac muscle,inhibiting vascular smooth muscle proliferation,lowering bloodpressure,reduction of blood lipid,lowering blood sugar level,anti-platelet aggregation,etc.Inrecent years,the antioxidant activity and antitumor activity researches of berberine attractedmuch attentions.To study the antioxidant activity of berberine,the MDA determination,agarose gelelectrophoresis,comet assay and SDS-PAGE were used to detect the protections against lipidperoxidation,DNA and protein oxidative degradation caused by free radicals.The xanthineoxidase oxidase - NBT reduction method,methyl violet,DMPD?+ and Oyaiuz method,wereused for the detection of the scavenging abilities of berberine to superoxide anions,hydroxylradicals,DMPD?+ free radicals and the reducing power.The Cytotoxicities and Inhibitions toliver cancer cells HepG2 and human erythroleukemia cells K562 of berberine were tested.TheDNA damage and apoptosis in K562 cells induced by berberine were also detected.Theresults are as follows:(1) Berberine was effective in preventing oxidative damage on biologicalmacromoleculars.The inhibitive rate to the lipid peroxidation was 1.6 % at the concentrationof 5μmol·L-1and was 95.5 % at 320μmol·L-1.The protection was slight at low concentration(5μmol·L-1) in oxidative plasmid pBR322DNA system triggered by AAPH at 10 mmol·L-1andapparently protective effect at 80μmol·L-1.The DNA damage in human peripheral bloodmononuclear cells induced by 50μmol·L-1H2O2 decreased when protected by 6.25μmol·L-1 ofberberine, the damage rate reduced to 66.2% from 81.7%(positive control).When theconcentration reached 100μmol·L-1,the protective effect was very strong,the damage ratedecreased to 48.4 %.The protective effect against the oxidation of protein was close to Troloxand appeared complete protection at 320μmol·L-1.(2) Berberine exhibited strong scavenging effects on the superoxide radical and hydroxylradical,its effect was more efficient thanTrolox.The percentage scavenging of berberine tosuperoxide radical was 99.2 % at the concentration of 160μmol·L-1.The scavenging ability to hydroxyl radical was as strong as Trolox,The scavenging rat for berberine and Trolox was91.0 % and 96.0 % respectively at 160μmol·L-1.Its ability to scavenge DMPD·+ was weak inoxidative DMPD system triggered by FeCl3 the scavenging rate only was 35.0 % at 1 280μmol·L-1 .In addition,Oyaiuz method showed that the reducing power of berberine was better.(3) Berberine exhibited strong inbition on the liver cancer cells HepG2 and humanerythroleukemia cells K562 and.the antiproliferations in both cells were in time- andconcentration- dependent. The IC50 value of berberine was 19μmol·L-1 and 9μmol·L-1 inHepG2 at 24 , 48 h respectively. The IC50 value for K562 was 11μmol·L-1 and 3μmol·L-1respectively. Therefore, in both cells berberine exihibited higher cytotoxicity.Moreover,K562cells were more sensitive to berberine than HepG2 cells. Both cells killed by berberine athigher concentration of 80,160μmol·L-1at 48 h.(4) The DNA damage in K562 cells initiated by berberine was detected using cometassay.The results showed that the damage rate was 23.4 % (The control was 10.0 % ) at theconcentration of 2.5μmol·L-1 at 48 h. The damage rate had reached 92.8% at concentration of160μmol·L-1. That showed berberine could induce serious DNA damage in K562 cells at highconcentrations.(5) The apoptosis in K562 cells induce by berberine was also investigated by AO/EBstaining assay.The apoptosis is in a time- and dose- dependent pattern.The rate were 16.9 %and 29 % at 40μmol·L-1 and 160μmol·L-1 respectively.The apoptosis rate reached 75.2 % at160μmol·L-1 at 48 h.In conclusion,berberine were effective in many oxidative systems and had great effectson growth inhibition in two cancer cell lines.In addition,it could induce K562 apoptosis athigher concentration. The apoptosis may be connected with the cell DNA damage.
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