Vol.:(0123456789) 1 3 Neurochem Res DOI 10.1007/s11064-017-2403-9 ORIGINAL PAPER Diferential Molecular Targets for Neuroprotective Efect of Chlorogenic Acid and its Related Compounds Against Glutamate Induced Excitotoxicity and Oxidative Stress in Rat Cortical Neurons Olfa Rebai 1  · Manel Belkhir 1  · María Victoria Sanchez‑Gomez 3  · Carlos Matute 3  · Sami Fattouch 2  · Mohamed Amri 1   Received: 21 June 2017 / Revised: 19 August 2017 / Accepted: 11 September 2017 © Springer Science+Business Media, LLC 2017 protective mechanism. Interseling, CGA and CA also exhibit antiapoptotic properties against glutamate-induced cleaved activation of pro-caspases; caspase 1,8 and 9 and calpain (PD 150606,Calpeptin and MDL 28170).These data sug- gest that neuroprotective activity of CGA ester may occurs throught its hydrolysate,the cafeic acid and its interaction with intracellular molecules suggesting that CGA exert its neuroprotection via its cafeoly acid group that might poten- tially be used as a therapeutic agent in neurodegeneratives disorders associated with glutamate excitotoxicity. Keywords Glutamate · Neuroprotection · Chlorogenic acid · Antioxidant · Apoptosis · Molecular mechanisms Abbreviations CNS Central nervous system DCF Dichlorofuorescein FDA Fluorescein diacetate H 2 O 2 Hydrogen peroxide LDH Lactate dehydrogenase PKA Protein kinase A PKC Protein kinase C PLC Phospholipase C ROS Reactive oxygen species MAPK Mitogen apoptotic protein kinase DPPH 2, 2-diphenyl-1-picrylhydrazyl DCF 2’, 7’-dichlorofuorescein CGA Chlorogenic acid CA Cafeic QA Quinic acid PBS Phosphate-bufered saline LDH Lactate deshydrogenase CAT Catalase SOD Superoxide dismutase NMDA N-methyl-D-aspartate Abstract The present study has been designed to explore the molecular mechanism and signaling pathway targets of chlorogenic acid (CGA) and its main hydrolysates, caf- feic (CA) and quinic acid in the protective efect against glutamate-excitotoxicity. For this purpose 8-DIV cortical neurons in primary culture were exposed to 50 μM L-glu- tamic acid plus 10 µM glycine, with or without 10–100 μM tested compounds. Chlorogenic acid and cafeic acid via their antioxidant properties inhibited cell death induced by glutamate in dose depended manner. However, quinic acid slightly protects neurons at a higher dose. DCF, JC-1 and Ca 2+ sensitive fuorescent dye fura-2, were used to measure intracellular ROS accumulation, mitochondrial membrane potential integration and intracellular calcium concentra- tion [Ca 2+ ] i . Results indicate that similarly, CGA acts as a protective agent against glutamate-induced cortical neu- rons injury by suppressing the accumulation of endogenous ROS and restore the mitochondrial membrane potential, activate the enzymatic antioxidant system by the increase levels of SOD activity and modulate the rise of intracel- lular calcium levels by increasing the rise of intracellular concentrations of Ca 2+ caused by glutamate overstimula- tion. PKC signaling cascade appear to be engaged in this * Olfa Rebai olfa.rebai@yahoo.fr 1 Research Unit of Functional Neurophysiology and Pathology, 00/UR/08-01, Department of Biological Sciences, Faculty of Science of Tunis, University of Tunis El Manar, 2092 Tunis, Tunisia 2 Laboratoire de Biochimie Alimentaire, INSAT, University of Carthage, Tunis, Tunisia 3 Departamento de Neurociencias, Facultad de Medicina y Odontologia, Universidad Del Paıs Vasco, Leioa, Vizcaya, Spain