Abstract
Lethal autophagy is a pathway leading to neuronal death caused by transient global ischemia. In this study, we examined the effect of Ginsenoside Rb1 (GRb1) on ischemia/reperfusion-induced autophagic neuronal death and investigated the role of PI3K/Akt. Ischemic neuronal death in vitro was induced by using oxygen glucose deprivation (OGD) in SH-SY5Y cells, and transient global ischemia was produced by using two vessels occlusion in rats. Cellular viability of SH-SY5Y cells was assessed by MTT assay, and CA1 neuronal death was evaluated by Hematoxylin-eosin staining. Autophagic vacuoles were detected by using both fluorescent microscopy in combination with acridine orange (AO) and Monodansylcadaverine (MDC) staining and transmission electronic microscopy. Protein levels of LC3II, Beclin1, total Akt and phosphor-Akt at Ser473 were examined by western blotting analysis. GRb1 inhibited both OGD and transient ischemia-induced neuronal death and mitigated OGD-induced autophagic vacuoles in SH-SY5Y cells. By contrast, PI3K inhibitor LY294002 counteracted the protection of GRb1 against neuronal death caused by either OGD or transient ischemia. LY294002 not only mitigated the up-regulated protein level of phosphor Akt at Ser473 caused by GRb1, but also reversed the inhibitory effect of GRb1 on OGD and transient ischemia-induced elevation in protein levels of LC3II and Beclin1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Apoptosis Regulatory Proteins / biosynthesis
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Apoptosis Regulatory Proteins / genetics
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Autophagy / drug effects*
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Beclin-1
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Brain Ischemia / drug therapy*
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Brain Ischemia / pathology
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CA1 Region, Hippocampal / blood supply
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CA1 Region, Hippocampal / pathology*
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Cell Line, Tumor
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Chromones / pharmacology
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Enzyme Activation / drug effects
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Ginsenosides / antagonists & inhibitors
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Ginsenosides / pharmacology
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Ginsenosides / therapeutic use*
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Glucose / pharmacology
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Humans
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Male
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Microtubule-Associated Proteins / biosynthesis
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Microtubule-Associated Proteins / genetics
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Morpholines / pharmacology
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Nerve Tissue Proteins / biosynthesis
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Nerve Tissue Proteins / genetics
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Neuroblastoma / pathology
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Neurons / drug effects*
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Neurons / metabolism
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Neurons / ultrastructure
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Neuroprotective Agents / antagonists & inhibitors
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Neuroprotective Agents / pharmacology
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Neuroprotective Agents / therapeutic use*
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Oxygen / pharmacology
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Phosphatidylinositol 3-Kinases / physiology*
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Phytotherapy*
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Proto-Oncogene Proteins c-akt / physiology*
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Random Allocation
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Rats
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Rats, Wistar
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Reperfusion Injury / drug therapy
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Reperfusion Injury / etiology
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Reperfusion Injury / prevention & control
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Signal Transduction / drug effects*
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Up-Regulation / drug effects
Substances
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Apoptosis Regulatory Proteins
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Beclin-1
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Becn1 protein, rat
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Chromones
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Ginsenosides
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LC3 protein, rat
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Microtubule-Associated Proteins
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Morpholines
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Nerve Tissue Proteins
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Neuroprotective Agents
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2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
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ginsenoside Rb1
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Phosphatidylinositol 3-Kinases
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Proto-Oncogene Proteins c-akt
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Glucose
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Oxygen