Ferulic acid protects PC12 neurons against hypoxia by inhibiting the p-MAPKs and COX-2 pathways

Document Type: Original Article


1 Department of Optometry, Yuanpei University, Hsinchu, Taiwan

2 Department of Physical Education Office, Yuanpei University, Hsinchu, Taiwan

3 Department of Biotechnology, Yuanpei University, Hsinchu, Taiwan


Objective(s):Hypoxia induces cellular oxidative stress that is associated with neurodegenerative diseases. Here, the protective effects of ferulic acid (FA) on hypoxia-induced neurotoxicity in PC12 cells were evaluated.
Materials and Methods:We investigated the effect of FA on PC12 cells subjected to hypoxia stress, in vitro.
Results:FA increased cell viability, prevented membrane damage (LDH release), scavenged free radicals, increased superoxide dismutase (SOD) activity, and attenuated the elevation of intracellular free Ca2+, lipid peroxidation, apoptosis (evaluated by TUNEL staining) and PGE2 production in hypoxia-stressed PC12 cells. MAPKs were activated during hypoxia. FA reduced p-p38 MAPK, caspase-3, and COX-2 activation which correlated well with diminished LDH release in PC12 cells under hypoxia. Furthermore, FA reduced lipid peroxidation in PC12 cells subjected to hypoxia.
Conclusion:Taken together, these results indicate that FA antioxidant effects could partly be involved in inhibition of p38 MAPK pathway and apoptosis through scavenging ROS in hypoxia-stressed PC12 cells.


1. Almeida A, Delgado-Esteban M, Bolanos JP, Medina JM. Oxygen and glucose deprivation induces mitochondrial dysfunction and oxidative stress in neurons but not in astrocytes in primary culture. J Neurochem 2002; 81:207-217.

2. Lievre V, Becuwe P, Bianchi A. Intracellular generation of free radicals and modifications of detoxifying enzymes in cultured neurons from the developing rat forebrain in response to transient hypoxia. Neuroscience 2001; 105:287-297.

3. Gonzalez-Perez O, Gonzalez-Castaneda RE, Huerta M, Luquin S, Gomez-Pinedo U, Sanchez-Almaraz E,et al. Beneficial effects of alpha-lipoic acid plus vitamin E on neurological deficit, reactive gliosis and neuronal remodeling in the penumbra of the ischemic rat brain. Neurosci Lett 2002; 321:100-104.

4. Ikeda K, Negishi H, Yamori Y. Antioxidant nutrients and hypoxia/ischemia brain injury in rodents. Toxicology 2003; 189:55-61.

5. Wang J, Lou J, Luo C. Phenolic Compounds from halimodendron halodendron (Pall.) voss and their antimicrobial and antioxidant activities. Int J Mol Sci 2012; 13:11349-11364.

6. Mori H, Kawabata K, Yoshimi N, Tanaka T, Murakami T, et al. Chemopreventive effects of ferulic acid on oral and rice germ on large bowel carcinogenesis. Anticancer Res 1999; 19:3775-3778.

7. Kim HY, Park J, Lee KH, Lee DU, Kwak JH, Kim YS ,et al. Ferulic acid protects against carbon tetrachloride-induced liver injury in mice. Toxicology 2011; 282:104-111.

8. Hou RC, Huang HM, Tzen JT, Jeng KC. Protective effects of sesamin and sesamolin on hypoxic neuronal and PC12 cells. J Neurosci Res 2003; 74:123-133.

9. Wang MJ, Lin WW, Chen HL, Chang YH, Ou HC, Kuo JS, et al. Silymarin protects dopaminergic neurons against lipopolysaccharide-induced neurotoxicity by inhibiting microglia activation. Eur J Neurosci 2002; 16:2103-2112.

10. Nozaki K, Nishimura M, Hashimoto N. Mitogen-activated protein kinases and cerebral ischemia. Mol Neurobiol 2001; 23:1-19.

11. Regan RF, Wang Y, Ma X, Chong A, Guo Y. Activation of extracellular signal-regulated kinases potentiates hemin toxicity in astrocyte cultures. J Neurochem 2001; 79:545-555.

12. Franke WW, Grund C, Achtstätter T. Co-expression of cytokeratins and neurofilament proteins in a permanent cell line: cultured rat PC12 cells combine neuronal and epithelial features. J Cell Biol 1986; 103:1933-1943.

13. Drukarch B, Schepens E, Stoof JC, Langeveld CH, Van Muiswinkel FL.  Astrocyte-enhanced neuronal survival is mediated by scavenging of extracellular reactive oxygen species. Free Radic Biol Med 1998; 25:217-220.

14. Hwang SL, Yen GC. Neuroprotective effects of the citrus flavanones against H2O2-induced cytotoxicity in PC12 cells. J Agric Food Chem 2008; 56:859-864.

15. Paul A, Belton A, Nag S, Martin I, Grotewiel MS, Duttaroy A. Reduced mitochondrial SOD displays mortality characteristics reminiscent of natural aging.Mech Ageing Dev 2007; 128:706-716.

16. Chao CC, Hu S, Peterson PK. Activated microglia mediate neuronal cell injury via a nitric oxide mechanism. J Immunol 1992; 149:2736-2741.

17. Zhang ZT, Cao XB, Xiong N, Wang HC, Huang JS, et al. Morin exerts neuroprotective actions in Parkinson disease models in vitro and in vivo. Acta Pharmacol Sin 2010; 31:900-906.

18. Huang HM, Ou HC, Hsieh SJ. Antioxidants prevent amyloid peptide-induced apoptosis and alteration of calcium homeostasis in cultured cortical neurons. Life Sci 2000; 66:1879-1892.

19. Floyd RA. Antioxidants oxidative stress and degenerative neurological disorders. Proc Soc Exp Biol Med 1999; 222:236-245.

20. Homi HM, Freitas JJ, Curi R, Velasco IT, Junior BA. Changes in superoxide dismutase and catalase activities of rat brain regions during early global transient ischemia/reperfusion. Neurosci Lett 2002; 333:37-40.

21. Wang CC, Fang KM, Yang CS, Tzeng SF. Reactive oxygen species-induced cell death of rat primary astrocytes through mitochondria-mediated mechanism. J Cell Biochem 2009; 107:933-943.

22. Lennon AM, Ramauge M, Dessouroux A, Pierre M. MAP kinase cascades are activated in astrocytes and preadipocytes 12–14 by 15-deoxy-prostaglandin J2 and the thiazolidinedione ciglitazone through peroxisome proliferator activator receptor gamma-independent mechanisms involving reactive oxygenated species. J Biol Chem 2002; 277:29681-29685.

23. Legos JJ, McLaughlin B, Skaper SD, Strijbos PJ, Parsons AA, et al. The selective p38 inhibitor SB-239063 protects primary neurons from mild to moderate excitotoxic injury. Eur J Pharmacol 2002; 447:37-42.

24. Tabakman R, Jiang H, Levine RA, Kohen R, Lazarovici P. Apoptotic characteristics of cell death and the neuroprotective effect of homocarnosine on pheochromocytoma PC12 cells exposed to ischemia. J Neurosci Res 2004; 75:499-507.

25. Jiang J, Borisenko GG, Osipov A, Martin I, Chen R, Shvedova AA, et al. Arachidonic acid-induced carbon-centered radicals and phospholipid peroxidation in cyclo-oxygenase-2-transfected PC12 cells. J Neurochem 2004; 90:1036-1049.

26. Yoo KY, Hwang IK, Kim JD, Kang IJ, Park J, et al. Antiinflammatory effect of the ethanol extract of Berberis koreana in a gerbil model of cerebral ischemia/reperfusion. Phytother Res 2008; 22:1527-1532.