s-Methyl cysteine enhanced survival of nerve growth factor differentiated PC12 cells under hypoxic conditions.
A nerve growth factor-differentiated PC12 cell line was used to investigate the protective effects of s-methyl cysteine (SMC) at 1, 2, 4, and 8 μM under oxygen-glucose deprivation (OGD) conditions. OGD decreased the cell viability. However, SMC pre-treatments at 2, 4 and 8 μM improved the cell viability, decreased cleaved caspase-3 and Bax expression, and reserved Bcl-2 expression. Furthermore, SMC maintained the mitochondrial membrane potential, lowered the intracellular Ca(2+) concentration and DNA fragmentation, and decreased the activity and expression of caspase-3 and caspase-8. OGD increased the reactive oxygen species (ROS) and 3-nitrotyrosine production, decreased glutathione peroxide (GPX) and glutathione reductase (GR) activities and the expression, enhanced nitric oxide synthase (NOS) activity and inducible NOS (iNOS) expression. SMC pre-treatments at 2, 4 and 8 μM lowered the ROS and 3-nitrotyrosine formation, maintained GPX and GR activities and expression, and decreased NOS activity and iNOS expression. OGD up-regulated hypoxia-inducible factor (HIF)-1α, nuclear transcription factor kappa (NF-κ) B p50, NF-κB p65 and p-p38 expression. SMC pre-treatments at 1-8 μM lowered HIF-1α expression and decreased p38 phosphorylation. SMC at 2, 4 and 8 μM suppressed the protein expression of NF-κB p50 and NF-κB p65. When YC-1 (HIF-1α inhibitor), pyrrolidine dithiocarbamate (NF-κB inhibitor) or SB203580 (p38MAPK inhibitor) were used to block the activation of HIF-1α, NF-κB and p38, SMC pre-treatments did not affect the protein expression of HIF-1α, NF-κB and p-p38. These results indicated that SMC was a potent neuro-protective agent.
Department of Neurosurgery, China Medical University Hospital, Taichung City, Taiwan.
Chun-Lin
CL
Liu
Te-Chun
TC
Hsia
Mei-Chin
MC
Yin
D000818:Animals; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002470:Cell Survival / Q000187:drug effects; D003545:Cysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005979:Glutathione Peroxidase / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D007191:Indazoles / Q000494:pharmacology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D020932:Nerve Growth Factor / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D016716:PC12 Cells; D011725:Pyridines / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D013859:Thiocarbamates / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D015854:Up-Regulation; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism
C494828:Bax protein, rat; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007093:Imidazoles; D007191:Indazoles; D016328:NF-kappa B; D018696:Neuroprotective Agents; D011725:Pyridines; D011759:Pyrrolidines; D017382:Reactive Oxygen Species; D013859:Thiocarbamates; D051028:bcl-2-Associated X Protein; C090937:3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole; C020972:pyrrolidine dithiocarbamic acid; C002744:3-nitrotyrosine; D014443:Tyrosine; D020932:Nerve Growth Factor; C008425:S-methylcysteine; D005979:Glutathione Peroxidase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D005980:Glutathione Reductase; D048051:p38 Mitogen-Activated Protein Kinases; C505410:Casp3 protein, rat; C505528:Casp8 protein, rat; D053148:Caspase 3; D053181:Caspase 8; D003545:Cysteine; C093642:SB 203580