AIMTo investigate the protective effect of monosialoganglioside (GM1) on injury induced by oxygen glucose deprivation/reperfusion (OGD/Rep) in rat hippocampal slices.

METHODSThe protective effects of GM1 on hippocampal slices after OGD/Rep were observed by detecting the light transmittance (LT) changes of rat hippocampal slices and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining of rat hippocampal slices.

RESULTS(1) In four groups treated with 0 (control), 0.1, 1.0, 10 micromol/L GM1, the peak of light transmittance (LT) in the slices treated with 1.0 micromol/L GM1 was significantly lower than that of the control and the group treated with 0.10 micromol/L GM1 (P < 0.01, ANOVA), while the peak of LT in the slices treated with 10.0 micromol/L GM1 was significantly lower than that of the other groups (P < 0.01, ANOVA). The time to reach the peak of LT in four groups was significantly different from each other (P < 0.05, Kruskal-Wallis test). The time to reach the peak of LT in the group treated with 1 micromol/L GM1 was the significantly longer than that in the control (P < 0.01, Mann-Whitney U test). (2) There was characteristic dose-response relationship between GM1 and TTC staining of rat hippocampal slices. In the five groups, treated with 0 (control), 0.01, 0.1, 1.0, 10 micromol/L GM1 respectively, TTC staining in the group treated with 1 micromol/L GM1 was the deepest (P < 0.05 vs. control, 0.01 and 0.1 micromol/L GM1 group, ANOVA), and the next was in the group treated with 10 micromol/L GM1 (P < 0.05 vs. control and 0.01 micromol/L GM1 group, ANOVA).

CONCLUSIONGM1 could protect injury induced by OGD/Rep in rat hippocampal slices effectively in vitro.

Animals ; G(M1) Ganglioside ; pharmacology ; Glucose ; deficiency ; Hippocampus ; metabolism ; Hypoxia ; metabolism ; In Vitro Techniques ; Male ; Oxygen ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury

Human neuroblastoma SH-SY5Y cell is a cloned cell line which has many attractive features for the study of neuronal proliferation and neurite outgrowth, because it has receptors for insulin, IGF-I and PDGF. Gangliosides are sialic acid containing glycosphingolipids which form an integral part of the plasma membrane of many mammalian cells. They inhibit cell growth mediated by tyrosine kinase receptors and ligand-stimulated tyrosine kinase activity, and autophosphorylation of EGF(epidermal growth factor) and PDGF receptors. The experiment was designed to study the effects of GM1 ganglioside on growth of human neuroblastoma SH-SY5Y cells stimulated with trophic factor in vitro. The cells were plated in Eagle's minimum essential medium without serum. The number and morphologic change of SH-SY5Y cells were evaluated in the serum free medium added GM1 ganglioside with insulin or PDGF. SH-SY5Y cells were maintained for six days in serum-free medium, and then cultured for over two weeks in serum-free medium containing either insulin or PDGF. The effect of insulin on cell proliferation developed earlier and was more potent than that of PDGF. These proliferative effects were inhibited by GM1 ganglioside, and the cells showed prominent neurites outgrowth. These findings suggest that GM1 ganglioside inhibits the cell proliferation mediated by tyrosine kinase receptors and directly induces neuritogenesis as one of the neurotrophic factors.
Cell Differentiation/drug effects ; Cell Division/drug effects ; G(M1) Ganglioside/*pharmacology ; Humans ; Insulin/pharmacology ; Neuroblastoma ; Neurons/cytology/*drug effects ; Platelet-Derived Growth Factor/pharmacology ; Receptor Protein-Tyrosine Kinases/drug effects ; Tumor Cells, Cultured

OBJECTIVETo study the effects of ganglioside 1 (GM1) and nerve growth factor (NGF) on neural stem cells (NSCs) proliferation in vitro.

METHODSNSCs were isolated and cultured in vitro. NSCs were cultured in the medium containing epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) or without the two agents. Different concentrations of GM1 and NGF were added into the two different medium. MTT and cell ball counting methods were used to ascertain the proliferation of NSCs. Immunohistochemical technology was used to observe the effect of GM1 and NGF on the proliferation of NSCs.

RESULTSHigh concentrations of GM1 (100 ng/L and 200 ng/L) promoted significantly the proliferation of NSCs in the medium containing EGF and bFGF (p<0.05). In the differentiation medium containing serum but no EGF and bFGF, NSCs proliferation increased with increasing concentration of GM1; the proportion of neurons and gliacytes increased with increasing concentration of NGF.

CONCLUSIONSHigh concentration of GM1 can promote NSCs proliferation and NGF can promote NSCs differentiation.

Animals ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Female ; G(M1) Ganglioside ; pharmacology ; Intermediate Filament Proteins ; analysis ; Male ; Nerve Growth Factors ; pharmacology ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; drug effects ; physiology

OBJECTIVETo investigate the effects of GM1 on inducing adult rat bone marrow stromal cells (MSCs) to form neural progenitor cells and their differentiation.

METHODSPurified MSCs were induced by different components of basic fibroblast growth factor (bFGF) alone, GM1 alone or combination of bFGF with GM1. After 3 days' incubation, fibronectin and collagen I were detected with immunocytochemistry, and nestin was detected with immunofluorescence. Neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and galactose cerebroside (GalC) were detected with immunocytochemistry after 7 days' incubation.

RESULTSAfter induction with bFGF alone or combination of bFGF and GM1, some MSCs exhibited the phenotypes of neural progenitor cells, and then neurons and astrocytes. In these two groups, the positive cells for fibronectin and collagen I decreased markedly after 3 days' induction. At the same time, the positive cells for nestin increased markedly. After 7 days' induction, NSE and GFAP-positive cells increased significantly. Furthermore, the addition of bFGF and GM1 caused the maximal variation. However, addition of GM1 alone had no inductive effects.

CONCLUSIONSCombination of bFGF with GM1 may synergistically promote the transformation of MSCs and differentiation into neurons and astrocyte-like cells. The results suggest a promising route for the application of MSCs.

Analysis of Variance ; Animals ; Bone Marrow Cells ; Cell Differentiation ; drug effects ; physiology ; Cells, Cultured ; Drug Synergism ; Fibroblast Growth Factor 2 ; pharmacology ; Fluorescent Antibody Technique ; G(M1) Ganglioside ; pharmacology ; Immunohistochemistry ; Probability ; Rats ; Rats, Wistar ; Sensitivity and Specificity ; Stem Cells ; pathology ; physiology ; Stromal Cells ; drug effects ; physiology ; ultrastructure

OBJECTIVETo determine the protective effect of monosialoganglionside (GM1) and evaluate the influence of GM1 on expression of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) in Sprague-Dawley (SD) rats with focal cerebral ischemia-reperfusion (I/R).

METHODSLeft middle cerebral artery (MCA) was occluded by an intraluminal suture for 1 h and the brain was reperfused for 72 h in SD rats when infarct volume was measured, GM1 (10 mg/kg) was given ip (intraperitoneally) at 5 min (group A), 1 h (group B) and 2 h (group C) after MCA occlusion (MCAo). Expression of NMDAR1 was detected by Western blot at various time after reperfusion (4 h, 6 h, 24 h, 48 h and 72 h) in ischemic hemispheres of the rats with or without GM1 administered.

RESULTS(1) Adjusted relative infarct volumes of groups A and B were significantly smaller than that of group C and the control group (P<0.01 and P<0.05, respectively). (2) Expression level of NMDAR1 was temporally high at 6 h after reperfusion, and dipped below the normal level at 72 h after reperfusion. GM1 at 5 min after MCAo significantly suppressed the expression of NMDAR1 at 6 h after reperfusion (P<0.05 vs the control). At 72 h after reperfusion, the NMDAR1 expression level of rats treated with GM1 administered (at 5 min or 2 h after MCAo) was significantly higher than that of the control (P<0.05).

CONCLUSIONGM1 can time-dependently reduce infarct volume in rats with focal cerebral I/R partly through stabilizing the expression of NMDAR1.

Animals ; Brain Ischemia ; metabolism ; pathology ; prevention & control ; G(M1) Ganglioside ; pharmacology ; therapeutic use ; Gene Expression Regulation ; drug effects ; Male ; Middle Cerebral Artery ; surgery ; Neurons ; drug effects ; physiology ; Protein Subunits ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Treatment Outcome