Hypoxia-inducible factor 1 （HIF-1） attenuates amyloid-beta protein neurotoxicity and decreases apoptosis induced by oxidative stress or hypoxia in cortical neurons. In this study, we construct-ed a recombinant adeno...Hypoxia-inducible factor 1 （HIF-1） attenuates amyloid-beta protein neurotoxicity and decreases apoptosis induced by oxidative stress or hypoxia in cortical neurons. In this study, we construct-ed a recombinant adeno-associated virus （rAAV） vector expressing the human HIF-1αgene （rAAV-HIF-1α）, and tested the assumption that rAAV-HIF-1αrepresses hippocampal neuronal apoptosis induced by amyloid-beta protein. Our results conifrmed that rAAV-HIF-1αsigniifcant-ly reduces apoptosis induced by amyloid-beta protein in primary cultured hippocampal neurons. Direct intracerebral rAAV-HIF-1αadministration also induced robust and prolonged HIF-1αproduction in rat hippocampus. Single rAAV-HIF-1αadministration resulted in decreased apoptosis of hippocampal neurons in an Alzheimer＇s disease rat model established by intrace-rebroventricular injection of aggregated amyloid-beta protein （25-35）. Our in vitro and in vivo ifndings demonstrate that HIF-1 has potential for attenuating hippocampal neuronal apoptosis induced by amyloid-beta protein, and provides experimental support for treatment of neurode-generative diseases using gene therapy.展开更多
By study on the effect of anisodamine on lipopolysaccharide- induced expression of tissue factor(TF) in vascular endothelial cells(EC) ,the mechanism of anisodam ine antithrom bosis,as well as in the treatm ent of b...By study on the effect of anisodamine on lipopolysaccharide- induced expression of tissue factor(TF) in vascular endothelial cells(EC) ,the mechanism of anisodam ine antithrom bosis,as well as in the treatm ent of bacteraemic shock was investigated.Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion m ethod.TF activity was measured in the lysates of HUVEC by using a single step clotting assay.Specific m RNA expression was detected by Northern blotting.In order to evaluate a possible contribution of the nuclear factor (NF) -κB pathway on the effects observed,electrophoretic mobility shift assays(EMSA) were performed using nuclear extracts from HU VECs and NF- κB- binding oligonucleotides.The results showed that treatment of HUVEC with L PS resulted in a significant increase in TF activity.Anisodamine dose- dependently inhibited L PS- induced upregulation of TF.These effects was also confirm ed on the level of specific TF m RNA expression by Northern blotting.Furtherm ore,EMSA showed that anisodamine com pletely abolished L PS- induced NF-κB DNA binding activity in nuclear ex- tracts from HUVECs treated with L PS together with anisodamine.The results suggest thataniso- damine counteracts endothelial cell activation by inhibiting L PS- induced TF expression in these cells.Its interference with the NF- κB pathway might- at least in part- contribute to this effect. The ability of anisodamine to counteract L PS effect on endothelial cells m ight be one underlying m echanism explaining its antithrombosis and efficacy in the treatment of bacteraemic shock.展开更多
基金supported by the National Natural Science Foundation of China,No.81273983the Natural Science Foundation of Hebei Province of China,No.C2010001471the Scientific Research Fund of Hebei Provincial Education Department in China,No.Q2012036
文摘Hypoxia-inducible factor 1 （HIF-1） attenuates amyloid-beta protein neurotoxicity and decreases apoptosis induced by oxidative stress or hypoxia in cortical neurons. In this study, we construct-ed a recombinant adeno-associated virus （rAAV） vector expressing the human HIF-1αgene （rAAV-HIF-1α）, and tested the assumption that rAAV-HIF-1αrepresses hippocampal neuronal apoptosis induced by amyloid-beta protein. Our results conifrmed that rAAV-HIF-1αsigniifcant-ly reduces apoptosis induced by amyloid-beta protein in primary cultured hippocampal neurons. Direct intracerebral rAAV-HIF-1αadministration also induced robust and prolonged HIF-1αproduction in rat hippocampus. Single rAAV-HIF-1αadministration resulted in decreased apoptosis of hippocampal neurons in an Alzheimer＇s disease rat model established by intrace-rebroventricular injection of aggregated amyloid-beta protein （25-35）. Our in vitro and in vivo ifndings demonstrate that HIF-1 has potential for attenuating hippocampal neuronal apoptosis induced by amyloid-beta protein, and provides experimental support for treatment of neurode-generative diseases using gene therapy.
基金This projectwas supported byagrantfrom National Natu-ral Sciences Foundation of China (No. 39730 2 2 0 ) andWuhan Chenguang Program (No. 985 0 0 30 86)
文摘By study on the effect of anisodamine on lipopolysaccharide- induced expression of tissue factor(TF) in vascular endothelial cells(EC) ,the mechanism of anisodam ine antithrom bosis,as well as in the treatm ent of bacteraemic shock was investigated.Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion m ethod.TF activity was measured in the lysates of HUVEC by using a single step clotting assay.Specific m RNA expression was detected by Northern blotting.In order to evaluate a possible contribution of the nuclear factor (NF) -κB pathway on the effects observed,electrophoretic mobility shift assays(EMSA) were performed using nuclear extracts from HU VECs and NF- κB- binding oligonucleotides.The results showed that treatment of HUVEC with L PS resulted in a significant increase in TF activity.Anisodamine dose- dependently inhibited L PS- induced upregulation of TF.These effects was also confirm ed on the level of specific TF m RNA expression by Northern blotting.Furtherm ore,EMSA showed that anisodamine com pletely abolished L PS- induced NF-κB DNA binding activity in nuclear ex- tracts from HUVECs treated with L PS together with anisodamine.The results suggest thataniso- damine counteracts endothelial cell activation by inhibiting L PS- induced TF expression in these cells.Its interference with the NF- κB pathway might- at least in part- contribute to this effect. The ability of anisodamine to counteract L PS effect on endothelial cells m ight be one underlying m echanism explaining its antithrombosis and efficacy in the treatment of bacteraemic shock.
