ABSTRACT
Objective To investigate the effect of sulforaphane on 1-methyl-4-phenylpyridinium (MPP +)-induced cell viability loss in cultured PC12 cells and to explore the possible mechanism.Methods MPP + induced damage in PC12 cells was prepared as oxidative damage model.Sulforaphane (0.5,1.O,2.5,5.0 and 10.0 μmol/L) was added in each group cell growth medium.Subsequent experiments were divided into 4 groups:(A) normal control group,(B) sulforaphane group,(C) MPP+ injury group,(D)sulforaphane pretreatment + MPP+ injury group.Cell viability was detected by MTT assay,and the sulforaphane pretreatment PC12 cell viability was observed in different concentrations.Flow cytometry was used to detect changes in the rate of apoptosis in different packet PC12 cells,and protein expression levels of nuclear factor erythroid2-related factor 2 (Nrf2),heme oxygenase (HO-1) and human NAD (P) H dehydrogenase,quinone 1 (NQO1) were detected by Western blot when the PC12 cells were incubated with sulforaphane (2.5 μmol/L) and (or) MPP+ (500 μmol/L) for 24 h in vitro.Results Compared to control group (cell survival rate was 98.70％),the survival percents of PC12 cells were significantly decreased in MPP+-treated group (58.16％).A significant difference was showed between group A and C (F =21.83,P ＜ 0.05),and the cell survival rate in group D was significantly improved.Compared to control group,the rate of apoptosis in MPP+ injury group was increased,and the rate of apoptosis after pretreatment of the sulforaphane was significantly reduced.Compared to MPP+ injury group,the levels of Nrf2,HO-1 and NQO1 protein expression were significantly increased in sulforaphane pretreatment group.Conclusion Sulforaphane have a protective effect against MPP+-induced PC12 cell model damage,and the protective effect may be achieved by activating the Nrf2-antioxidant response element pathway.
ABSTRACT
Objective To develop transgenic mice harboring the fusion gene of mutant amyloid precursor protein and two types of fluorescent protein for the future study on Alzheimer's disease.Methods The fusion gene CFP-54 bp-YFP-C99 was introduced into mice by mieroinjection.The presence of CFP-54 bp-YFP-C99 was confirmed by PCR in the founders.Results CFP-54 bp-YFP-C99 gene was injected into pronucleus of 2202 zygotes and 1806 injected eggs were implanted into 56 foster mothers, 13 of which were pregnant.There were 13 foster mothers who borne 52 offspring and 32 of them survived.Recipient mouse pregnancy rate was 23.2% (13/56) and the integration rate was 3.9% (2/52).Conclusion CFP-54 bp-YFP-C99 transgenic mice is obtained, but the transgenic efficiency is low.