Résumé
Objective To observe the protective effect of recombinant plasmid pCDNA3.1(+)/Ngb during focal cerebral ischemia in rat brain.Methods Fifty-four male Wistar rats were randomly divided into three groups:normal saline(NS)control group,plasmid control group,and recombinant neuroglobulin group.NS,plasmid pCDNA3.1(+)and recombinant plasmid pCDNA3.1(+)/Ngb were respectively injected into two sites of the rat cerebra1 cortex 24 hours before induction of neocortical focal ischemia by occlusion of the right middle cerebral artery for 24 hours.The condition of local ischemic damage,expression of bcl-2 and the apoptosis in neural cells were confirmed by staining with 2% 2.3.5-triphenyltetrazolium chloride,in-site cell apoptosis detection,indirect immunofluorescent staining and Western blotting,respectively.Results The extent of cerebral infarction tissue and the apoptosis cells in the pCDNA3.1(+)/Ngb group were significantly reduced than those in other control groups(P
Résumé
Objective To clone rat neuroglobulin (NGB) gene and construct its eukaryotic expression vector. Methods The total RNA was extracted from Wistar rat brain and the full length cDNA encoding NGB was obtained by RT-PCR. After the sequence was confirmed by sequencing and BLAST, it was inserted in the eukaryotic expression vector pCDNA3.1 (+), then the sequence and reading frame were confirmed by two restriction endonucleases and sequencing. Results The NGB gene was cloned with four bases mutated and its eukaryotic expression vector was constructed. Conclusion NGB expressed in Wistar rat brain. NGB gene was successfully cloned and inserted in eukaryotic expression vector.