ABSTRACT
Objective: To establish an oxygen and glucose deprivation model (OGD) of SH-SY5Y cells, and investigate the effects of Xuesaitong Injection on the cell survival, apoptosis rate of SH-SY5Y cells and the mRNA and protein expressions of Lingo-1. Methods: The model was established using 1640 sugar-free medium and three-gas incubator. The cell survival rate was determined by CCK8 method to determine the optimal time for hypoxia and the optimal concentration of Xuesaitong Injection. The apoptosis rate of SH-SY5Y cells was detected by Annexin V-FITC/PI double staining. The mRNA and protein expressions of Lingo-1 were determined by qRT-PCR and Western blotting, respectively, and the effect of Xuesaitong on Lingo-1 expression was finally determined. Results: In this study, the optimal hypoxia time for the establishment of the OGD model of SH-SY5Y cells was 16 h and the optimum concentration of Xuesaitong Injection was 640 mg/L. The apoptosis rate of SH-SY5Y cells was significantly reduced, and the expression levels of lingo-1 mRNA and protein were decreased in the Xuesaitong group compared with the model group under the condition of this concentration. Conclusion: The apoptosis rate was significantly increased and Lingo-1 was highly expressed when SH-SY5Y cells were damaged by oxygen and glucose deprivation. Xuesaitong can significantly reduce the apoptosis of SH-SY5Y cells induced by oxygen and glucose deprivation, and inhibit the high expression of lingo-1, which has the anti-apoptosis and significant neuroprotective effect.
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Objective To detect the expressions of Lingo-1 and myelin associated protein in the white matter and hippocampus of ovariectomized rats after short-term estrogen replacement therapy in order to explore the possible mechanisms for the effects of estrogen on the brain myelin sheaths and cognitive function.Methods 24 middleaged (9-12 months) female Sprague-Dawley (SD) rats were bilaterally ovariectomized (OVX) and randomly divided into vehicle replacement (OVX+Veh) group and estrogen replacement (OVX+E) group.After one month ERT,The spatial learning and memory ability of all rats were assessed with Morris water maze.Then,10 rats were randomly selected from each group.The ultrastucture of myelin sheaths in the cerebral white matter and hippocampus were observed,and the protein expression of MBP and Lingo-1 were investigated with Western blot and immunohistochemical staining.Results The escape latencies of OVX+E rats in navigation test were significantly shorter than that of OVX+Veh rats(P＜0.05).The myelin sheaths in the white matter and hippocampus of OVX+Veh rats showed obviously degeneration.In the OVX+E group,the expression of MBP in the white matter and hippocampus was significantly higher than that of OVX+Veh group(P＜0.05),however,the expression of Lingo-1 was significantly lower than that of OVX +Veh group (P＜ 0.05).Conclusions One-month ERT has significant beneficial effects on the spatial learning capacity and myelin sheaths in the white matter and hippocampus.The protective effects may be related to estrogen-induced downregulation of the Lingo-1 expression in the white matter and hippocampus of rats.
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Objective To explore the effect of exercise on the expression of the LINGO-1 gene after intra cerebral hemorrhage (ICH).Methods Seventy-two Sprague-Dawley rats were randomly divided into a sham operation group,a control group and an exercise training group,each of 24.The three groups were further divided into 1 d,3 d,7 d and 14 d groups.ICH was induced using intra-parenchymal injection of autologous blood.The exercise training group was then forced to exercise on a treadmill.Any recovery of neurological functional was evaluated using Longa scoring,and the expression of LINGO-1 mRNA and protein were detected using the reverse transcription-polymerase chain reaction and western blotting.Results The sham operation group displayed no obvious neurological deficiency,with little expression of LINGO-1 mRNA or protein at any time point.The control group's average Longa score reached a maximum of 3 seven days after the operation,decreasing to 2 after another 7 days.The average expression of LINGO-1 mRNA and protein in that group peaked at 1.335±0.393 three days after the ICH,then decreased to 0.429±0.035 on the 7th day and 0.371±0.038 on the 14th day.In the exercise training group the average Longa score on the 7th day was 2,the average LINGO-1 mRNA level was 0.257±0.042 and the average protein level was 1.142±0.287,all significantly lower than in the control group.Moreover,in the exercise group there was a significant positive correlation between LINGO-1 protein expression and the Longa scores.Conclusion Exercise can decrease the expression of LINGO-1 mRNA and protein and promote the recovery of neural function after ICH.
ABSTRACT
Objective To explore the effect of exercise on the expression of the LINGO-1 gene after intra cerebral hemorrhage (ICH).Methods Seventy-two Sprague-Dawley rats were randomly divided into a sham operation group,a control group and an exercise training group,each of 24.The three groups were further divided into 1 d,3 d,7 d and 14 d groups.ICH was induced using intra-parenchymal injection of autologous blood.The exercise training group was then forced to exercise on a treadmill.Any recovery of neurological functional was evaluated using Longa scoring,and the expression of LINGO-1 mRNA and protein were detected using the reverse transcription-polymerase chain reaction and western blotting.Results The sham operation group displayed no obvious neurological deficiency,with little expression of LINGO-1 mRNA or protein at any time point.The control group's average Longa score reached a maximum of 3 seven days after the operation,decreasing to 2 after another 7 days.The average expression of LINGO-1 mRNA and protein in that group peaked at 1.335±0.393 three days after the ICH,then decreased to 0.429±0.035 on the 7th day and 0.371±0.038 on the 14th day.In the exercise training group the average Longa score on the 7th day was 2,the average LINGO-1 mRNA level was 0.257±0.042 and the average protein level was 1.142±0.287,all significantly lower than in the control group.Moreover,in the exercise group there was a significant positive correlation between LINGO-1 protein expression and the Longa scores.Conclusion Exercise can decrease the expression of LINGO-1 mRNA and protein and promote the recovery of neural function after ICH.
ABSTRACT
Three myelin proteins, Nogo-A, MAG and OMgp, transduce their neurite-outgrowth inhibitory signal through a common receptor complex: NgR/ p75NTR (or TROY). Recently, LINGO-1 is identified as another essential component and regulator for the Nogo-66 receptor/p75 signaling complex. LINGO-1 is restricted to express in CNS, neuronal LINGO-1 is shown to be involved in the signal transduction from three myelin proteins, and Lingo-1 in oligodendrocyte negatively regulates the differentiation and myelination of oligodendrocyte. To investigate the potential activity of LINGO-1 in neuronal apoptosis, LINGO-1-Fc fusion protein including the extracellular LRR and IgC2 domain, was used as functional antagonist to study its protective effect on low-potassium induced apoptosis of cerebellar granule neurons (CGNs). In judgement of the apoptotic nuclei stained by Hoechst, LINGO-1-Fc pretreatment for 2 h significantly prevents apoptosis of CGNs. Although GST-LINGO-1 protein, including the LRR domain, binds to the CGN cultures in the same way with LINGO-1-Fc, it doesn't prevent the apoptosis of CGNs. These results indicate that LINGO-1-Fc fusion protein prevents low-potassium induced apoptosis of cerebellar granule neurons in certain conditions and this activity is probably IgC2 domain dependent.