Hyperhomocysteinemia(HHcy)is independently associated with poorer long-term prognosis in patients with intracerebral haemorrhage(ICH);however,the effect and mechanisms of HHcy on ICH are still unclear.Here,we evaluate...Hyperhomocysteinemia(HHcy)is independently associated with poorer long-term prognosis in patients with intracerebral haemorrhage(ICH);however,the effect and mechanisms of HHcy on ICH are still unclear.Here,we evaluated neurite outgrowth and neurological functional recovery using simulated models of ICH with HHcy in vitro and in vivo.We found that the neurite outgrowth velocity and motor functional recovery in the ICH plus HHcy group were significantly slower than that in the control group,indicating that homocysteine(Hcy)significantly impedes the neurite outgrowth recovery after ICH.Furthermore,phosphoproteomic data and signalome analysis of perihematomal brain tissues suggested that calmodulin-dependent protein kinases 2(CAMK2A)kinase substrate pairs were significantly downregulated in ICH with HHcy compared with autologous blood injection only,both western blot and immunofluorescence staining confirmed this finding.Additionally,upregulation of pCAMK2A significantly increased neurite outgrowth recovery in ICH with HHcy.Collectively,we clarify the mechanism of HHcy-hindered neurite outgrowth recovery,and pCAMK2A may serve as a therapeutic strategy for promoting neurological recovery after ICH.展开更多
Background:Ferrous ion,a degradation product of hematomas,induces inflammatory reactions and other secondary injuries after intracerebral hemorrhage(ICH).Our study aimed to investigate the specific neuroprotective mec...Background:Ferrous ion,a degradation product of hematomas,induces inflammatory reactions and other secondary injuries after intracerebral hemorrhage(ICH).Our study aimed to investigate the specific neuroprotective mechanism of adipose-derived stem cells(ADSCs)on ferrous ion-induced neural injury in vitro.Methods:ADSCs were co-cultured with primary cortical neurons in a transwell system treated with ferrous sulfate to generate an in vitro ICH model.ADSCs and cortical neurons were cultured in the upper and lower chambers,respectively.Neuron apoptosis was determined by flow cytometry.The levels of insulin-like growth factor-1(IGF-1),malondialdehyde(MDA)and nitric oxide synthase(NOS)activity in neuron culture medium were detected with commercial kits.In neurons,protein expression in phosphatidylinositol-3-kinase(PI3 K)/protein kinase B(Akt)signaling pathway,nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway and apoptosis-related proteins were detected by western blot.Results:ADSCs attenuated neural apoptosis,reduced MDA levels and NOS activity induced by ferrous sulfate.In neurons,IGF-1 was increased,as were p-PI3 K,p-Akt,Nrf2,HO-1,and Bcl-2 while cleaved caspase 3 was down-regulated.Conclusions:ADSCs exert neuroprotective effects against ferrous iron-induced neuronal damage by secreting IGF-1 and increasing the levels of Akt-dependent Nrf2/ARE signaling pathway.展开更多
文摘Hyperhomocysteinemia(HHcy)is independently associated with poorer long-term prognosis in patients with intracerebral haemorrhage(ICH);however,the effect and mechanisms of HHcy on ICH are still unclear.Here,we evaluated neurite outgrowth and neurological functional recovery using simulated models of ICH with HHcy in vitro and in vivo.We found that the neurite outgrowth velocity and motor functional recovery in the ICH plus HHcy group were significantly slower than that in the control group,indicating that homocysteine(Hcy)significantly impedes the neurite outgrowth recovery after ICH.Furthermore,phosphoproteomic data and signalome analysis of perihematomal brain tissues suggested that calmodulin-dependent protein kinases 2(CAMK2A)kinase substrate pairs were significantly downregulated in ICH with HHcy compared with autologous blood injection only,both western blot and immunofluorescence staining confirmed this finding.Additionally,upregulation of pCAMK2A significantly increased neurite outgrowth recovery in ICH with HHcy.Collectively,we clarify the mechanism of HHcy-hindered neurite outgrowth recovery,and pCAMK2A may serve as a therapeutic strategy for promoting neurological recovery after ICH.
基金supported by grants from the National Natural Science Foundation of China to Prof.Zhouping Tang(Grant Nos.81171089 and 81471201)
文摘Background:Ferrous ion,a degradation product of hematomas,induces inflammatory reactions and other secondary injuries after intracerebral hemorrhage(ICH).Our study aimed to investigate the specific neuroprotective mechanism of adipose-derived stem cells(ADSCs)on ferrous ion-induced neural injury in vitro.Methods:ADSCs were co-cultured with primary cortical neurons in a transwell system treated with ferrous sulfate to generate an in vitro ICH model.ADSCs and cortical neurons were cultured in the upper and lower chambers,respectively.Neuron apoptosis was determined by flow cytometry.The levels of insulin-like growth factor-1(IGF-1),malondialdehyde(MDA)and nitric oxide synthase(NOS)activity in neuron culture medium were detected with commercial kits.In neurons,protein expression in phosphatidylinositol-3-kinase(PI3 K)/protein kinase B(Akt)signaling pathway,nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway and apoptosis-related proteins were detected by western blot.Results:ADSCs attenuated neural apoptosis,reduced MDA levels and NOS activity induced by ferrous sulfate.In neurons,IGF-1 was increased,as were p-PI3 K,p-Akt,Nrf2,HO-1,and Bcl-2 while cleaved caspase 3 was down-regulated.Conclusions:ADSCs exert neuroprotective effects against ferrous iron-induced neuronal damage by secreting IGF-1 and increasing the levels of Akt-dependent Nrf2/ARE signaling pathway.
