| Abstract |
The present study is to study drug metabolic characteristics of CYP3A46
gene. Sus scrofa CYP3A46 gene from Bama miniature pig liver was obtained
by PCR method and cloned in pCDNA3.1, the recombinant plasmid pCDNACYP3A46
was obtained. The recombinant plasmid and empty plasmid were transfected
into HepG2 which designated as HepG2-CYP3A46 and HepG2-CDNA3.1 repectively.
The stably expressing cell lines HepG2-CYP3A46 and HepG2-pcDNA3.1 were
achieved by the selection of gentamycin (G418) ,and were incubated with
the probe drug nifedipine (NF) in optimal conditions. After the cells were
incubated for 60 minutes, the high performance liquid chromatography
(HPLC) was utilized to detect the change of metabolites in cells. The
results showed that the sequence of cloned Sus scrofa CYP3A46 gene was
correct, and the stably expressing cell the cell lines transfected with
pCDNA-CYP3A46 and pCDNA3.1 were established successfully. The activity
analysis demonstrated that the cell line transfected with pCDNA-CYP3A46
had the significantly metabolic activities of nifedipine( P<0.001).
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