Lee, SM Yune, TY Kim, SJ Kim, YC Oh, YJ Markelonis, GJ Oh, TH 2024-01-21T06:08:44Z 2024-01-21T06:08:44Z 2021-09-05 2004-11 0022-3042 https://pubs.kist.re.kr/handle/201004/137102 We attempted to ascertain the neuroprotective effects and mechanisms of minocycline in inflammatory-mediated neurotoxicity using primary neuron/glia co-cultures treated with lipopolysaccharide (LPS). Neuronal cell death was induced by treatment with LPS for 48 h, and the cell damage was assessed using lactate dehydrogenase (LDH) assays and by counting microtubule-associated protein-2 (MAP-2) positive cells. Through terminal transferase deoxyuridine triphosphate-biotin nick end labeling (TUNEL)-staining and by measuring caspase-3 activity, we found that LPS-induced neuronal cell death was mediated by apoptosis. We determined that pre-treatment with minocycline significantly inhibited LPS-induced neuronal cell death. In addition, LPS induced inducible nitric oxide synthase (iNOS) expression significantly, resulting in nitric oxide (NO) production within glial cells, but not in neurons. Both nitric oxide synthase (NOS) inhibitors (N-G-monomethyl-L-arginine monoacetate (L-NMMA) and S-methylisothiourea sulfate (SMT)) and minocycline inhibited iNOS expression and NO release, and increased neuronal survival in neuron/glia co-cultures. Pre-treatment with minocycline significantly inhibited the rapid and extensive production of tumor necrosis factor-alpha (TNF-alpha) mediated by LPS in glial cells. We also determined that the signaling cascade of LPS-mediated iNOS induction and NO production was mediated by TNF-alpha by using neutralizing antibodies to TNF-alpha. Consequently, our results show that the neuroprotective effect of minocycline is associated with inhibition of iNOS induction and NO production in glial cells, which is mediated by the LPS-induced production of TNF-alpha. English WILEY NITRIC-OXIDE SYNTHASE NECROSIS-FACTOR-ALPHA ACTIVATED PROTEIN-KINASES SPINAL-CORD-INJURY MICROGLIAL ACTIVATION INTERFERON-GAMMA NITROGEN-OXIDES MESSENGER-RNA MOUSE MODEL CYTOKINE Minocycline inhibits apoptotic cell death via attenuation of TNF-alpha expression following iNOS/NO induction by lipopolysaccharide in neuron/glia co-cultures Article 10.1111/j.1471-4159.2004.02780.x 1 JOURNAL OF NEUROCHEMISTRY, v.91, no.3, pp.568 - 578 JOURNAL OF NEUROCHEMISTRY 91 3 568 578 scie scopus 000224524900006 2-s2.0-7244242682 Biochemistry & Molecular Biology Neurosciences Biochemistry & Molecular Biology Neurosciences & Neurology Article NITRIC-OXIDE SYNTHASE NECROSIS-FACTOR-ALPHA ACTIVATED PROTEIN-KINASES SPINAL-CORD-INJURY MICROGLIAL ACTIVATION INTERFERON-GAMMA NITROGEN-OXIDES MESSENGER-RNA MOUSE MODEL CYTOKINE inducible nitric oxide synthase lipopolysaccharide minocycline neuronal cell death nitric oxide TNF-alpha