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Studies have shown that acetylcholinesterase inhibitors donepezil and galantamine have effects of reducing neuronal damage caused by glucose deprivation and reducing the cerebral infarction volume of cerebral ischemic animals, but their effects may not be entirely dependent on its inhibition of cholinesterase activity. In order to study the effects of donepezil and galantamine on neuronal injury of cerebral ischemia, the rat neuron-astrocyte co-culture model was successfully established in this study. In this model, we studied the effects of donepezil and galantamine on neuron apoptosis induced by oxygen-glucose deprivation/reoxygenation (OGD/R) and investigated the mechanism. The results showed that donepezil and galantamine significantly reduced the neuron apoptosis, and promoted the synthesis and secretion of BDNF and NGF in astrocytes in the co-culture system. Donepezil and galantamine activated the PI3K/Akt pathway and ERK pathway, and promoted the phosphorylation of the nuclear transcription factor CREB. These results suggest that donepezil and galantamine exhibit protective effects on neuronal damage induced by OGD/R. The mechanism may be related to activation of PI3K/Akt pathway and ERK pathway in astrocytes and promote phosphorylation of CREB, which lead to the synthesis and secretion of BDNF and NGF from astrocytes.
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Objective To investigate the effects of Zuogui Jiangtang Jieyu Prescription on neurotrophic effects of astrocytes in diabetes mellitus rats with depression. Methods The diabetes mellitus with depression rat models were established by composite method, and then were randomly divided into 5 groups: model group, positive group, Zuogui Jiangtang Jieyu Prescription high-, medium-, low-dose groups, with 12 rats in each group. 12 normal rats were set as control group. Each administration group was given relevenat medicine for gavage for continuous 4 weeks. Open-field test was used to evaluate the depression-like behavior. The expression of GFAP in astrocyte and MAP2 in neuron were tested by immunohistochemistry. The expression of protein and mRNA of BDNF, GDNF, and NGF were tested by Western blot and RT-PCR. Results Compared with the control group, the activities of rats in model group were significantly reduced. The expression of GFAP increased, while the expressions of MAP2, BDNF, GDNF and NGF decreased. Compared with the model group, the depression-like behavior of rats in model group were significantly improved. The expression of GFAP decreased, while the expressions of MAP2, BDNF, GDNF and NGF increased, and GFAP decrseaed significantly. Conclusion The secretion function of astrocyte can be improved by Zuogui Jiangtang Jieyu Prescription. Its anti-depression and neuron-protection function might be correlated with the enhancement of neurotrophic effects of astrocytes.
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@#Objective To investigate the effect of Morroniside which was isolated from Cornus officinalis on the growth and the protection against H2O2-induced injury in SH-SY5Y neuroblastoma cells.Methods SH-SY5Y cells were incubated with Morroniside in different concentration.MTT metabolic rate was measured as the cell survival rate in order to determine the best effective concentration to promote the survival of neuron.MTT metabolic rate,axonal length,area of cell body and cell counting were used as the indicators to analyze the effects of Morroniside on the growth of SH-SY5Y on the best effective concentration.After SH-SY5Y cells were injured with 500 μmol/L H2O2,they were administrated with Morroniside in different concentration and vitamin E.The expression of neural growth factor(NGF)were determined with Western Blotting.Results Morroniside increased the cell survival rate in 10～100 μmol/L.Compared with control group,MTT metabolic rate(P<0.05),axonal length(P<0.001),area of cell body(P<0.01),cell counting(P<0.05)increased in morroniside-treated group after SH-SY5Y cells were incubated with Morroniside in 10 μmol/L.The expression of NGF in morroniside-treated group was higher than that of the control group after injury induced by H2O2.Conclusion Morroniside shows the neurotrophic effect and can protect SH-SY5Y cells from injury induced by H2O2.
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Objective To analyze the function of cGMP-dependent protein kinase(PKG)in the survival of cultured rat cerebellar granule cells(CGC).Methods CGC culture was incubated for 24 h before addition of ethanol and different reagents,such as Deta-NONOate(a NO donor),Br-cGMP and Rp-8-pCPT-cGMPS(a cGMP-dependent protein kinase inhibitor).After the treatment,samples were incubated for another 24 h,then the cells were collected and counted by using hemocytometer.Results Deta-NONOate and Br-cGMP enhanced the cell survival in the absence of ethanol,indicating their neurotrophic effects.They showed neuroprotective effect on the cultured CGC since they could reduce the ethanol-induced cell death.Inhibiting PKG with Rp-8-pCPT-cGMPS could eliminate both neurotrophic and neuroprotective effects mediated by Deta-NONOate and Br-cGMP.Conclusion cGMP-dependent protein kinase plays a key role in the cultured CGC survival and protects the cells against ethanol neurotoxicity.
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Objective To analyse the molecular structure of Schwann cell derived neurotrophic protein (SDNP) by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) Methods The purity of SDNP was digested byTPCK trypsin and detected by MALDI TOF MS, mass mapping was determined and partial sequences of the protein have neen analyzed by the mass data of fragment ion peaks in the fragmentation analysis and structural TOF MS (FAST) spectra The primary partial structure of SDNP was identified by searching the protein databases Results The integrity SDNP in peak detected by MALDI TOF MS has 66?10 3 MW and higher purity, because no other peaks exists except double charge peak The mass mapping of many peptides was determined and 8 peptides of them have been retrieved in MS Fit database, there is not the same protein in database Hypothetical protein has 5 peptides homology with the sample (62%) FAST spectra of 2305 Da shows the primary partial structure of SDNP after searching the protein MS Tag database Conclusions The molecular weight of SDNP detected by MALDI TOF MS is 66?10 3, The sequence of partial amino acid is EPVKKVTNSRRAKRTKPNGHIAN