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.
KeywordsGlutamate Neuroprotection Chlorogenic acid Antioxidant Apoptosis Molecular mechanisms
Central nervous system
Protein kinase A
Protein kinase C
Reactive oxygen species
Mitogen apoptotic protein kinase
Hank’s Balanced Salt Solution
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).
- 7.Kim J, Lee S, Shim J, Kim HW, Kim J, Young JJ, Yang H, Park J, Choi SH, Yoon JH, Lee KW, Lee HJ (2012) Caffeinated coffee, decaffeinated coffee, and the phenolic phytochemical chlorogenic acid up-regulate NQO1 expression and prevent H2O2-induced apoptosis in primary cortical neurons. Neurochem Int 60:466–474CrossRefPubMedGoogle Scholar
- 19.Zhang YM, Bhavnani B (2005) Glutamate-induced apoptosis in primary cortical neurons is inhibited by equine estrogens via down-regulation of caspase-3 and prevention of mitochondrial cytochrome c release. BMC Neurosci 6:13. https://bmcneurosci.biomedcentral.com/articles/10.1186/1471-2202-6-13
- 21.Anggreani E, Lee CY (2017) Neuroprotective effect of chlorogenic acids against Alzheimer’s disease. Int J Food Sci Nutr Diet 6(1):330–337Google Scholar
- 44.Poręba M, Stróżyk A, Salvesen GS, Drąg M (2013) Caspase substrates and inhibitors. Cold Spring Harb Perspect Biol 5(8). doi: 10.1101/cshperspect.a008680
- 50.Wingrave JM, Schaecher KE, Sribnick EA, Wilford GG, Ray SK, Hazen-Martin DJ, Hogan EL, Banik L (2003) Early induction of secondary injury factors causing activation of calpain and mitochondria-mediated neuronal apoptosis following spinal cord injury in rats. J Neurosci Res 73(1):95–104CrossRefPubMedGoogle Scholar