A study of optimal concentration of brain tissue extract of hypoxia-preconditioned mice to enhance tolerance of differentiated PC12 cells to hypoxia
- VernacularTitle:缺氧预适应小鼠脑匀浆提取液增强分化PC12细胞缺氧耐受性的最适浓度研究
- Author:
Hong LI
- Publication Type:Journal Article
- Keywords:
Hypoxia;
Preconditioning;
Mouse;
Brain tissue extract;
Differentiated PC 12 cell
- From:
Journal of Chongqing Medical University
2003;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of brain tissue extract of hypoxia-preconditioned mice (BHP) on tolerance of NGF-induced differentiated PC12 cells to hypoxia and the optimal concentration for protective effect.Methods:The mice model of acute repetitive hypoxia was reproduced and BHP was prepared.BHP was added into the cultures of 4～5 day differentiated PC12 cells and the final protein concentrations of BHP were 1.0、10.0、100.0?g/ml(BHP groups) respectively.Brain tissue ex- tract of normal mice(BN) at the same three protein concentrations were used as controls(BN groups) and PBS solvent control groups were also prepared in which only PBS was added(PBS groups).Then differentiated PC12 cells were cultured in hypoxia (2%O_2).After 24,48,72 hours'hypoxia,colorimetric method(OD_(570nm)) of tetrazolium salt MTT staining was adopted to determine cell viability and lactate dehydrogenase(LDH) leakage assays were also conducted at the same time.Besides,apoptotic per- centages at late stage(72 hours' hypoxia) were detected.Results:After 24 hours' hypoxia,OD_(570nm) values of BHP groups with protein concentrations being 10.0、100.0?g/ml were significantly higher,but their LDH leakage percentages were significantly lower than those of BN groups with the same protein concentrations and of hypoxic PBS group.With hypoxia prolonging,BHP gradually lost the protective effect.At the time point of 72 hours' hypoxia,OD_(570nm) values of only BHP group at 100.0?g/ml protein concentration was significantly higher than that of the corresponding BN groups,its LDH leakage percentage and apop- totic percentage was significantly lower than those of the hypoxic PBS group.Conclusion:The effects of BHP on tolerance of differentiated PC12 cells to hypoxia depend on its concentrations and on its treating time.Besides,BHP at final concentration of 100.01?g/ml protein showed steady,prolonged protective effect.
