Molecular Mechanisms Responsible for Neuron-Derived Conditioned Medium (NCM)-Mediated Protection of Ischemic Brain.

Abstract

The protective value of neuron-derived conditioned medium (NCM) in cerebral ischemia and the underlying mechanism(s) responsible for NCM-mediated brain protection against cerebral ischemia were investigated in the study. NCM was first collected from the neuronal culture growing under the in vitro ischemic condition (glucose-, oxygen- and serum-deprivation or GOSD) for 2, 4 or 6 h. Through the focal cerebral ischemia (bilateral CCAO/unilateral MCAO) animal model, we discovered that ischemia/reperfusion (I/R)-induced brain infarction was significantly reduced by NCM, given directly into the cistern magna at the end of 90 min of CCAO/MCAO. Immunoblocking and chemical blocking strategies were applied in the in vitro ischemic studies to show that NCM supplement could protect microglia, astrocytes and neurons from GOSD-induced cell death, in a growth factor (TGFβ1, NT-3 and GDNF) and p-ERK dependent manner. Brain injection with TGFβ1, NT3, GDNF and ERK agonist (DADS) alone or in combination, therefore also significantly decreased the infarct volume of ischemic brain. Moreover, NCM could inhibit ROS but stimulate IL-1β release from GOSD-treated microglia and limit the infiltration of IL-β-positive microglia into the core area of ischemic brain, revealing the anti-oxidant and anti-inflammatory activities of NCM. In overall, NCM-mediated brain protection against cerebral ischemia has been demonstrated for the first time in S.D. rats, due to its anti-apoptotic, anti-oxidant and potentially anti-glutamate activities (NCM-induced IL-1β can inhibit the glutamate-mediated neurotoxicity) and restriction upon the infiltration of inflammatory microglia into the core area of ischemic brain. The therapeutic potentials of NCM, TGFβ1, GDNF, NT-3 and DADS in the control of cerebral ischemia in human therefore have been suggested and require further investigation.

• Authors: Chi-Hsin Lin, Chen-Hsuan Wang, Shih-Lan Hsu, Li-Ya Liao, Ting-An Lin, Chi-Mei Hsueh
• Journal: PloS one (PLoS One) Vol. 11 Issue 1 Pg. e0146692 ( 2016) ISSN: 1932-6203 [Electronic] United States
• PMID: 26745377 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Culture Media, Conditioned
• Culture Media, Serum-Free
• Glial Cell Line-Derived Neurotrophic Factor
• Nerve Growth Factors
• Neuroprotective Agents
• Reactive Oxygen Species
• Transforming Growth Factor beta1
• Extracellular Signal-Regulated MAP Kinases

Topics

• Animals
• Apoptosis
• Brain (blood supply, metabolism, pathology)
• Cell Movement
• Cell Survival
• Cells, Cultured
• Culture Media, Conditioned
• Culture Media, Serum-Free
• Drug Evaluation, Preclinical
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• Glial Cell Line-Derived Neurotrophic Factor (physiology)
• Infarction, Middle Cerebral Artery (drug therapy, metabolism, pathology)
• Male
• Microglia (physiology)
• Nerve Growth Factors (physiology)
• Neurons (metabolism)
• Neuroprotective Agents (administration & dosage)
• Oxidative Stress
• Primary Cell Culture
• Protective Factors
• Rats, Sprague-Dawley
• Reactive Oxygen Species (metabolism)
• Reperfusion Injury (prevention & control)
• Transforming Growth Factor beta1 (physiology)