Vahid Reza Askari1, Narges Fereydouni2, Vafa Baradaran Rahimi3, Nafiseh Askari4, Amir Hossein Sahebkar5, Pouria Rahmanian-Devin6, Alireza Samzadeh-Kermani7. 1. Student Research Committee, Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Neurogenic Inflammation Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: vahidrezaaskary@yahoo.com. 2. Student Research Committee, Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. 3. Student Research Committee, Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. 4. Student Research Committee, Sabzevar University of Medical Sciences, Sabzevar, Iran. 5. Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. 6. Student Research Committee, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. 7. Department of Chemistry, Faculty of Science, University of Zabol, Zabol, Iran. Electronic address: arsamzadeh@yahoo.com.
Abstract
BACKGROUND: Inflammation is a primary response to infection that can pathologically lead to various diseases including neurodegenerative diseases. The purpose of this study was to evaluate the effect of β-Amyrin, a naturally occurring pentacyclic triterpenoid compound, on inflammation induced by lipopolysaccharide (LPS) and interferone-γ (IFN-γ) in rat microglial cells. MATERIALS AND METHODS: Cytotoxicity of β-Amyrin (3-100) μM on microglial cells was evaluated using the MTT assay. Also, the protective effect of various β-Amyrin (2-16 μM) concentrations with LPS/IFN-γ-induced mice microglial cells was studied. The concentrations of TNF-α (Tumor Necrosis Factor-α), IL-1β (Interleukin-1β), IL-6 (Interleukin-6) and PGE-2 (Prostaglandin E2) were evaluated using ELISA. Gene expressions of TNF-α, IL-1β, IL-6, COX-2 (Cyclooxygenase-2), iNOS and arginase-1 were also evaluated using the Real-Time PCR method. Nitrite oxide and urea were measured using biochemical methods. RESULTS: The studied concentrations of β-Amyrin had no significant effects on the viability of microglial cells. Interestingly, β-Amyrin concentration dependently and significantly increased the reduced cell proliferation concerning to LPS/IFN-γ exposure (p < 0.001). The concentrations and expression levels of pro-inflammatory factors including TNF-α, IL-1β, IL-6, PGE-2, COX-2 were significantly reduced after β-Amyrin treatment in LPS/IFN-γ-induced microglial cells (p < 0.05-0.001). β-Amyrin also decreased the levels of nitric oxide, increased urea and down regulated the expression of nitric oxide synthesis while arginase-1 expression was enhanced (p < 0.001). The ratio of NO/urea and iNOS/Arg1 were also markedly increased in comparison to the LPS/IFN-g group (p < 0.001). CONCLUSION: β-Amyrin reduces inflammation in microglial cells and can be used as a potential anti-inflammatory agent in central nervous system neurodegenerative diseases such as Alzheimer and multiple sclerosis, by affecting the inflammatory cytokine and differentiation of microglia as resident CNS macrophages.
BACKGROUND:Inflammation is a primary response to infection that can pathologically lead to various diseases including neurodegenerative diseases. The purpose of this study was to evaluate the effect of β-Amyrin, a naturally occurring pentacyclic triterpenoid compound, on inflammation induced by lipopolysaccharide (LPS) and interferone-γ (IFN-γ) in rat microglial cells. MATERIALS AND METHODS:Cytotoxicity of β-Amyrin (3-100) μM on microglial cells was evaluated using the MTT assay. Also, the protective effect of various β-Amyrin (2-16 μM) concentrations with LPS/IFN-γ-induced mice microglial cells was studied. The concentrations of TNF-α (Tumor Necrosis Factor-α), IL-1β (Interleukin-1β), IL-6 (Interleukin-6) and PGE-2 (Prostaglandin E2) were evaluated using ELISA. Gene expressions of TNF-α, IL-1β, IL-6, COX-2 (Cyclooxygenase-2), iNOS and arginase-1 were also evaluated using the Real-Time PCR method. Nitrite oxide and urea were measured using biochemical methods. RESULTS: The studied concentrations of β-Amyrin had no significant effects on the viability of microglial cells. Interestingly, β-Amyrin concentration dependently and significantly increased the reduced cell proliferation concerning to LPS/IFN-γ exposure (p < 0.001). The concentrations and expression levels of pro-inflammatory factors including TNF-α, IL-1β, IL-6, PGE-2, COX-2 were significantly reduced after β-Amyrin treatment in LPS/IFN-γ-induced microglial cells (p < 0.05-0.001). β-Amyrin also decreased the levels of nitric oxide, increased urea and down regulated the expression of nitric oxide synthesis while arginase-1 expression was enhanced (p < 0.001). The ratio of NO/urea and iNOS/Arg1 were also markedly increased in comparison to the LPS/IFN-g group (p < 0.001). CONCLUSION: β-Amyrin reduces inflammation in microglial cells and can be used as a potential anti-inflammatory agent in central nervous system neurodegenerative diseases such as Alzheimer and multiple sclerosis, by affecting the inflammatory cytokine and differentiation of microglia as resident CNS macrophages.
Authors: Karen Jaqueline Paredes-Ruiz; Karla Chavira-Ramos; Mario Orozco-Morales; Cimen Karasu; Alexey A Tinkov; Michael Aschner; Abel Santamaría; Ana Laura Colín-González Journal: Neurotox Res Date: 2021-11-06 Impact factor: 3.911