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Differential Molecular Targets for Neuroprotective Effect of Chlorogenic Acid and its Related Compounds Against Glutamate Induced Excitotoxicity and Oxidative Stress in Rat Cortical Neurons

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The present study has been designed to explore the molecular mechanism and signaling pathway targets of chlorogenic acid (CGA) and its main hydrolysates, caffeic (CA) and quinic acid in the protective effect against glutamate-excitotoxicity. For this purpose 8-DIV cortical neurons in primary culture were exposed to 50 μM l-glutamic acid plus 10 µM glycine, with or without 10–100 μM tested compounds. Chlorogenic acid and caffeic 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 Ca2+sensitive fluorescent dye fura-2, were used to measure intracellular ROS accumulation, mitochondrial membrane potential integration and intracellular calcium concentration [Ca2+] i . Results indicate that similarly, CGA acts as a protective agent against glutamate-induced cortical neurons 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 intracellular calcium levels by increasing the rise of intracellular concentrations of Ca2+caused by glutamate overstimulation. PKC signaling cascade appear to be engaged in this 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 suggest that neuroprotective activity of CGA ester may occurs throught its hydrolysate,the caffeic acid and its interaction with intracellular molecules suggesting that CGA exert its neuroprotection via its caffeoly acid group that might potentially be used as a therapeutic agent in neurodegeneratives disorders associated with glutamate excitotoxicity.

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Central nervous system




Fluorescein diacetate

H2O2 :

Hydrogen peroxide


Lactate dehydrogenase


Protein kinase A


Protein kinase C


Phospholipase C


Reactive oxygen species


Mitogen apoptotic protein kinase


2, 2-diphenyl-1-picrylhydrazyl


2’, 7’-dichlorofluorescein


Chlorogenic acid




Quinic acid


Phosphate-buffered saline


Lactate deshydrogenase




Superoxide dismutase




Dimethyl sulfoxide


Hank’s Balanced Salt Solution


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This work was supported by the Research Unit 00-UR-08-01 University of Sciences, Tunis and by a grant from the Tunisian, Ministry of Higher Education and Scientific Research Tunisia. The authors would like to thank Prof. Matute Carlos from the Instituto Del pays Vasco, Spain for their supportive advices. We acknowledge the financial support from the Ministry of Higher Education and Scientific Research (Tunisia).

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Correspondence to Olfa Rebai.

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Rebai, O., Belkhir, M., Sanchez-Gomez, M.V. et al. Differential Molecular Targets for Neuroprotective Effect of Chlorogenic Acid and its Related Compounds Against Glutamate Induced Excitotoxicity and Oxidative Stress in Rat Cortical Neurons. Neurochem Res 42, 3559–3572 (2017).

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