Protective estrogen-like properties and mechanism of quercetin in rat cerebral cortex neurons

OBJECTIVE To investigate the effect of quercetin on primary cultured newborn rat cortex neuron cell which is estrogen depletion,and discuss the possible mechanism,to provide new ideas and strategies for developing a drug of neurodegenerative disease.METHODS Rat cortex neurons were isolated from one day old Sprague Dawley rats and treated with estrogen,quercetin and estrogen receptor antagonists(ICI182,780).Cell viability was determined by MTT assay,neurite outgrowth was measured by fluorescent microsope and estrogen receptors were determine by Western blot.RESULTS Quercetin functions like estrogen to increase cortex neuronal cell viability,the Que(50,100μmol·L^(-1))group compared with the control group could significantly improve the activity of the cortical neurons(P<0.05).It can also increase neurite out growth,the Que(50,100μmol·L^(-1))group significantly promoted the formation of synapse,most of the neurons were full,and the synapses of neurons became thick,growth,and connect to a dense neural network.And in the Western blot experiments,Que(50,100μmol·L^(-1))group could obviously increase the expression of estrogen receptor alpha protein,in addition,the neural protective effect of quercetin can be inhibited by ICI182,780.CONCLUSION Quercetin like estrogen can protected cortex neuronal and the effect of quercetin on cortex neuronal cells was mediated by estrogen receptor alpha.

Neuroprotective effect of quercetin on beta-amyloid-induced PC12 cells injury and potential related mechanism

OBJECTIVE To explore the estrogen-like neuroprotective effects and the related mechanism of quercetin by using PC12 cells induced with Aβ25-35,provided thought and strategy for the drug therapy of AD.METHODS Cells were cultured with Aβ25-35 for 24 h,17β-estradiol(0.1 μmol·L^(-1)),genistein(50 μmol·L^(-1)) and three different concentrations of quercetin(200 μmol·L^(-1),300 μmol·L^(-1) and400 μmol·L^(-1)) were respectively added after 24 h.The effects of quercetin on activity of AD model were tested by MTT.Immunohistochemical stain and Western blot were used to detect the expression of estrogen receptors alpha and beta,p-ERK1/2 and apoptosis related proteins.The mechanism of quercetin estrogen-like neuroprotective effects was detected using estrogen receptor antagonist ICI182,780 and MAPKK inhibitor U0126.RESULTS The results revealed thatthe toxic effects showed in a dose-dependent increase of Aβ25-35 on PC12 cells.Comparing with the control group,cells injury was observed after cultured with 10 μmol·L^(-1) Aβ25-35 for 24 h(P<0.01).The MTT results showed that 17β-estradiol,genistein and three different concentrations of quercetin could significantly enhance the cell survival rate compared with the model group(P<0.05).Compared with model group,Immunofluorescence and Western blot results show that quercetin could improve the estrogen receptor alpha and p-ERK1/2 protein expression(P<0.05),and the expression of estrogen receptor beta protein is increased without significant difference.And in the Western blot experiments,the ratio of Bcl-2 and Bax was increased and the expression of Caspase 3 was decreased(P<0.05).When estrogen receptor inhibition ICI182,780 were reduced,the expression of p-ERK1/2 was decreased(P<0.05) and the ratio of Bcl-2 and Bax was decreased,Caspase 3 protein expression was increased(P<0.05).In addition,pretreatment of cells with U0126 would reduce Bcl-2/Bax ratio and increase Caspase 3 protein expression increased(P<0.05).CONCLUSION Quercetin protected PC12 cells,which suffered from Aβ25-35-induced cytotoxicity and exert neuroprotective effects.The estrogen-like neuroprotective effect can reduce the apoptosis in the classic estrogen receptor pathway and MAPK pathway.And quercetin can also active MAPK signaling pathways by the mediation of estrogen receptor alpha.

Protective effect and its mechanism of codonopsis pilosula polysaccharide on memory consolidation disorder induced by cycloheximide in mice

OBJECTIVE To investigate the protective effect of Codonopsis Pilosula Polysaccharide(CPPS)on improving of the memory consolidation disorder induced by Cycloheximide and its possible mechanisms in mice.METHODS The mice was divided into five groups,as normal control group,cycloheximid model group,piracetam positive control group,CPPS 300 mg·kg^(-1) group,and CPPS150 mg·kg^(-1) group.The mice respectively were given saline,piracetam,and CPPS for 15 d.The memory consolidation disorder model in mice was established by ip.Cyclohexylamine,and orally administered CPPS(300 mg·kg^(-1) or 150 mg·kg^(-1))every day.Then experimental groups were subjected Morris Water Maze test.Western blotting analysis were used to analysis the expression of Ca MKⅡ/CREB signaling pathways.RESULTS Morris water maze experiment showed that cyclohexylamine can cause memory consolidation disorder(P<0.01),and giving piracetam and CPPS(300 mg·kg-1)can improve spatial memory impairment in mice(P<0.05,P<0.01).Western blotting experiment results show that compared with normal control group,Ca MKⅡand CREB contents of brain in model group mice had significant decreased(P<0.001);Compared with model group,Ca MKⅡand CREB contents of brain tissue in piracetam and CPPS groups increased significantly(P<0.05,P<0.01,P<0.001).CONCLUSION Cycloheximide can induce the memory consolidation disorder,and its effect in mice related to Ca MK/CREB signaling pathways.CPPS can improved this memory disorder by influence Ca MKⅡ/CREB signaling pathways.