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AIM:To explore an ideal method for extracting protoporphyrindisodium (PPN) from unanticoagulated animal blood,andto study the inhibitory effects of PPN on HBV-DNA duplicationand its cytotoxicity to 2.2.15 cell strain.METHODS:Protoporphyrin methyl ester and otherintermediate products were prepared with protohemeseparated from protein hydrolysates of coagulated animalblood,which were finally made into PPN and detectedquantitatively with an ultraviolet fluorescent analyzer.Ten μg/ml,20 μg/ml,40 μg/ml,80 μg/ml and 160 μg/ml ofPPN-aqueous solution were added into culture medium for2.2.15 cells respectively.Eight days later,the drugconcentration in supernatant from the culture medium wasdetected when inhibition rate of HBeAg,cell survival ratewhen inhibition rate of HBeAg was 50% (ID50),and whensurvival cells in experimental group were 50% of those incontrol group (CD50),and the therapeutic index (TI) wasalso detected.PPN with different concentration of 10 μg/ml,20 μg/ml,40 μg/ml,80 μg/ml and 160 μg/ml was respectivelymixed and cultivated with HepG2 2.2.15 cell suspension,and then the inhibition of PPN against HBV-DNA was judgedby PCR.RESULTS:The extract of henna crystal was identified to bePPN.When the concentrations of PPN were 160 μg/ml and80 μg/ml,the inhibition rates of HBeAg were 89.8% and82.4%,and the cell survival rates were 98.7% and 99.2%.CONCLUSION:It is suggested that PPN can be extractedfrom unanticoagulated animal blood.PPN can inhibit HBV-DNA expression and duplication in vitro,and has nocytotoxicity to liver cells.Further study and application ofPPN are warranted.
AIM: To explore an ideal method for extracting protoporphyrindisodium (PPN) from unanticoagulated animal blood, and to study the inhibitory effects of PPN on HBV-DNA duplication and its cytotoxicity to 2.2.15 cell strain. METHODS: Protoporphyrin methyl ester and other intermediate products were prepared with protohemeseparated from protein hydrolysates of coagulated animalblood, which were finally made into PPN and detectedquantitatively with an ultraviolet fluorescent analyzer. Ten μg / ml, 20 μg / ml, 40 μg / ml, 80 μg / ml and 160 μg / ml of PPN- were added into culture medium for 2.2.15 cells respectively. Eight days later, the drugconcentration in supernatant from the culture medium wasdetected when inhibition rate of HBeAg, cell survival rate inhibition inhibition of of HBeAg was 50% (ID50), and whensurvival cells in experimental group were 50% of those incontrol groups (CD50), and the therapeutic index (TI) was detected. PPN with different concentrations of 10 μg / ml, 20 μg / ml, 40 μg / ml, 80 μg / ml and 160 μg / ml were respectively cultivated with HepG2 2.2.15 cell suspension, and then the inhibition of PPN against HBV-DNA was judged by PCR .RESULTS: The extract of henna crystal was identified to be PPN. When the concentrations of The inhibition rates of HBeAg were 89.8% and 82.4%, and the cell survival rates were 98.7% and 99.2% respectively. CONCLUSION: It is suggested that PPN can extracted from unanticoagulated animal blood. PPN can inhibit HBV-DNA expression and duplication in vitro, and has nocytotoxicity to liver cells. Future study and application of PPN are warranted.