Neuroprotective Effects of Ethanol Extract of Polyscias fruticosa (EEPF) against Glutamate-Mediated Neuronal Toxicity in HT22 Cells

• Published in: International journal of molecular sciences 2023-02, Vol.24 (4), p.3969
• Main Authors: Selvaraj, Baskar, Le, Tam Thi, Kim, Dae Won, Jung, Bo Hyun, Yoo, Ki-Yeon, Ahn, Hong Ryul, Thuong, Phuong Thien, Tran, Thi Thu Thuy, Pae, Ae Nim, Jung, Sang Hoon, Lee, Jae Wook
• Format: Article
• Language: English
• Subjects: AKT protein; Animals; Apoptosis; Apoptosis-inducing factor; BAX protein; Bcl-2 protein; bcl-2-Associated X Protein - metabolism; Biocompatibility; Brain-derived neurotrophic factor; Brain-Derived Neurotrophic Factor - metabolism; Calcein; Calcium (intracellular); Calcium ions; Cell culture; Cell death; Cell Line; Cyclic AMP response element-binding protein; Dyes; Ethanol; Extracellular signal-regulated kinase; Flow cytometry; Fluo-3; Fluorescent dyes; Fluorescent indicators; Glutamic Acid - metabolism; Herbal medicine; Hippocampus - metabolism; HT22 cells; Intracellular; Investigations; Ischemia; Kinases; Magnoliopsida - chemistry; MAP kinase; Molecular modelling; Neurons - drug effects; Neuroprotection; Neuroprotective Agents - pharmacology; Oxidative Stress; Phosphorylation; Plant extracts; Plant Extracts - pharmacology; Polyscias fruticosa; Proteins; Proto-Oncogene Proteins c-akt - metabolism; reactive oxygen species; Reactive Oxygen Species - metabolism; Reagents; Toxicity; Transcription factors; Traumatic brain injury
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24043969

In traditional herbal medicine, the has been frequently used for the treatment of ischemia and inflammation. Oxidative stress mediated by elevated glutamate levels cause neuronal cell death in ischemia and various neurodegenerative diseases. However, so far, the neuroprotective effects of this plant extract against glutamate-mediated cell death have not been investigated in cell models. The current study investigates the neuroprotective effects of ethanol extracts of (EEPF) and elucidates the underlying molecular mechanisms of EEPFs relevant to neuroprotection against glutamate-mediated cell death. The oxidative stress-mediated cell death was induced by 5 mM glutamate treatment in HT22 cells. The cell viability was measured by a tetrazolium-based EZ-Cytox reagent and Calcein-AM fluorescent dye. Intracellular Ca and ROS levels were measured by fluorescent dyes, fluo-3 AM and 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA), respectively. Protein expressions of p-AKT, BDNF, p-CREB, Bax, Bcl-2, and apoptosis-inducing factor (AIF) were determined by western blot analysis. The apoptotic cell death was measured by flow cytometry. The in vivo efficacy of EEPF was evaluated using the Mongolian gerbil mouse by surgery-induced brain ischemia. EEPF treatment showed a neuroprotective effect against glutamate-induced cell death. The EEPF co-treatment reduced the intracellular Ca and ROS and apoptotic cell death. Furthermore, it recovered the p-AKT, p-CREB, BDNF, and Bcl-2 levels decreased by glutamate. The EEPF co-treatment suppressed the activation of apoptotic Bax, the nuclear translocation of AIF, and mitogen-activated protein kinase (MAPK) pathway proteins (ERK1/2, p38, JNK). Further, EEPF treatment significantly rescued the degenerative neurons in the ischemia-induced Mongolian gerbil in vivo model. EEPF exhibited neuroprotective properties that suppress glutamate-mediated neurotoxicity. The underlying mechanism of EEPF is increasing the level of p-AKT, p-CREB, BDNF, and Bcl-2 associated with cell survival. It has therapeutic potential for the treatment of glutamate-mediated neuropathology.