Chemical modification of ascorbic acid to L-ascorbyl-6-palmitate: A novel approach for improved antioxidant therapy in traumatic brain injury

Abstract

Oxidative stress, caused by an excessive amount of reactive oxygen species is a major factor in the pathophysiology of complications following traumatic brain injury (TBI). Ascorbic acid, a vital antioxidant, has been employed in TBI therapy, but its instability, limited bioavailability, rapid oxidation, and pro-oxidant effects pose significant limitations. To overcome these drawbacks, the ascorbic acid was chemically modified resulting in a fat-soluble L-ascorbyl-6-palmitate. The effects of L-ascorbyl-6-palmitate on oxidative stress biomarkers in TBI rats were subsequently evaluated. TBI was developed in rats by a weight drop method. The study involved five experimental groups: ascorbic acid group, L-ascorbyl-6-palmitate group, dimethyl sulfoxide group, traumatized non-treated group, and non-traumatized non-treated control group. A total of twenty-five rats were used in the experiment, with five rats in each group (n=5). The levels of malondialdehyde and the activity of antioxidant enzymes such as glutathione peroxidase, catalase, and superoxide dismutase were assessed in serum and brain tissue samples. In both serum and brain tissue, ascorbic acid, L-ascorbyl-6-palmitate, and dimethyl sulfoxide showed significant (P<0.05) elevation in enzyme activities and reduction in malondialdehyde levels compared to the traumatized non-treated group. Additionally, L-ascorbyl-6-palmitate treatment demonstrated higher antioxidant potential and scavenging ability than ascorbic acid treatment, as evidenced by significantly (P<0.05) increased superoxide dismutase, catalase, and glutathione peroxidase activities, and reduced malondialdehyde levels. These findings demonstrate the neuroprotective effects of L-ascorbyl-6-palmitate in managing TBI-induced oxidative stress. Further studies should investigate the underlying molecular mechanisms and long-term effects of L-ascorbyl-6-palmitate treatment on neurological recovery and functional outcomes in TBI, as well as explore its potential synergistic effects with other antioxidants or neuroprotective strategies.

Keywords

• Traumatic Brain Injury
• Oxidative stress
• Ascorbic Acid
• L-ascorbyl-6-palmitate
• Antioxidant enzymes

References

1. Orman JAL, Selassie AW, Perdue CL, Thurman DJ, Kraus JF. Surveillance of Traumatic Brain Injury. In: Li G, Baker S, editors. Injury Research. MA: Boston; Springer; 2012, p. 61-85.
2. Sivanandam TM, Thakur MK. Traumatic brain injury: a risk factor for Alzheimer's disease. Neurosci Biobehav Rev. 2012 May;36(5):1376-81.
3. Sahuquillo J, Poca MA, Amoros S. Current aspects of pathophysiology and cell dysfunction after severe head injury. Curr Pharm Des. 2001 Oct;7(15):1475-503.
4. Chan PH. Mitochondria and Neuronal Death/Survival Signaling Pathways in Cerebral Ischemia. Neurochem Res. 2004; 29(11): 1943-1949.
5. Xiong Y, Gu Q, Peterson P, Muizelaar J, Lee C. Mitochondrial Dysfunction and Calcium Perturbation Induced by Traumatic Brain Injury. J Neurotrauma. 1997; 14(1): 23-34.
6. Ames BN, Shigenaga MK, Hagen TM. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):7915-22.
7. Hanafy KA, Selim MH. Antioxidant Strategies in Neurocritical Care. Neurotherapeutics. 2011; 9(1): 44-55.
8. He L, He T, Farrar S, Ji L, Liu T, Ma X. Antioxidants Maintain Cellular Redox Homeostasis by Elimination of Reactive Oxygen Species. Cell Physiol Biochem. 2017; 44(2): 532-553.

9. Pun PBL, Lu J, Moochhala S. Involvement of ROS in BBB dysfunction. Free Radic Research. 2009; 43(4): 348-364.
10. Ishaq GM, Saidu Y, Bilbis LS, Muhammad SA, Jinjir N, Shehu BB. Effects of α-tocopherol and ascorbic acid in the severity and management of traumatic brain injury in albino rats. J Neurosci Rural Pract. 2013 Jul;4(3):292-7.
11. May JM. Vitamin C Transport and Its Role in the Central Nervous System. Subcell Biochem. 2011; 58: 85-103.
12. Harrison FE, May JM. Vitamin C function in the brain: vital role of the ascorbate transporter SVCT2. Free Radic Biol Med. 2009; 46(6): 719-730.
13. Santos Ítala MS, Tomé A da R, Saldanha GB, Ferreira PMP, Militão GCG, De Freitas RM. Oxidative Stress in The Hippocampus During Experimental Seizures Can Be Ameliorated with The Antioxidant Ascorbic Acid. Oxid Med Cell Longev. 2009; 2(4): 214-221.
14. Weber V, Coudert P, Rubat C, Duroux E, Leal F, Couquelet J. Antioxidant Properties of Novel Lipophilic Ascorbic Acid Analogues. J Pharm Pharmacol. 2000; 52(5): 523-530.
15. Tripathi R, Singh B, Bisht S, Pandey J. L-Ascorbic Acid in Organic Synthesis: An Overview. Curr Org Chem. 2009; 13(1): 99-122.
16. Bindhumol V, Chitra K, Mathur P. Ascorbic Acid induces reactive oxygen species generation in the liver of male rats. Toxicology. 2003; 188(2): 117-124.
17. Mohamed R, Dharmappa KK, Tarannum S, Jameel NM, Kannum SA, Ashrafulla HS, et al. Chemical modification of ascorbic acid and evaluation of its lipophilic derivatives as inhibitors of secretory phospholipase A2 with anti-inflammatory activity. Mol Cell Biochem. 2010; 345(1): 69-76.
18. Marmarou A, Foda MAA-E, Brink W van den, Campbell J, Kita H, Demetriadou K. A new model of diffuse brain injury in rats. J Neurosurg. 1994; 80(2): 291-300.
19. Rezanejad Keshteli FZ, Parivar K, Joghatayi MT, Beik HA. Study of the differentiation of rat omentum stem cells to nerve cells using brain tissue extract of Wistar rats. International J Cell Mol Biotechnol. 2014; 1-13.
20. Marklund SL. Analysis of extracellular superoxide dismutase in tissue homogenates and extracellular fluids. Methods Enzymol. 1990;186:260-5.
21. Johansson LH, Håkan Borg L. A spectrophotometric method for determination of catalase activity in small tissue samples. Anal Biochem. 1988; 174(1): 331-336.
22. Ursini F, Maiorino M, Gregolin C. The selenoenzyme phospholipid hydroperoxide glutathione peroxidase. Biochim Biophys Acta. 1985 Mar 29;839(1):62-70.
23. Goulart M, Batoréu M, Rodrigues A, Laires A, Rueff J. Lipoperoxidation products and thiol antioxidants in chromium exposed workers. Mutagenesis. 2005; 20(5): 311-315.
24. Rodriguez-Rodriguez A, Egea-Guerrero J, Murillo-Cabezas F, Carrillo-Vico A. Oxidative Stress in Traumatic Brain Injury. Curr Med Chem. 2014; 21(10): 1201-1211.
25. Verma RS, Mehta A, Srivastava N. In vivo chlorpyrifos induced oxidative stress: Attenuation by antioxidant vitamins. Pestic Biochem Physiol. 2007; 88(2): 191-196.
26. Pritam P, Deka R, Bhardwaj A, Srivastava R, Kumar D, Jha AK, Jha NK, Villa C, Jha SK. Antioxidants in Alzheimer’s Disease: Current Therapeutic Significance and Future Prospects. Biology. 2022; 11(2): 212.
27. Bulama I, Suleiman N, Bilbis L, Abbas A, Jinjiri N, Saidu Y, et al. Antioxidative and Neurotherapeutic Effect of Ascorbic acid on Albino rats Induced with Traumatic Brain Injury. J Cell Neurosci Oxid Stress. 2020; 12(1): 922-936.
28. Devi SA, Vani R, Subramanyam MVV, Reddy SS, Jeevaratnam K. Intermittent hypobaric hypoxia-induced oxidative stress in rat erythrocytes: protective effects of vitamin E, vitamin C, and carnitine. Cell Biochem Func. 2007; 25(2): 221-231.
29. Zaidi SMKR, Al-Qirim TM, Banu N. Effects of Antioxidant Vitamins on Glutathione Depletion and Lipid Peroxidation Induced by Restraint Stress in the Rat Liver. Drugs R D. 2005; 6(3): 157-165
30. Austria R, Semenzato A, Bettero A. Stability of vitamin C derivatives in solution and topical formulations. J Pharm Biomed Anal. 1997; 15(6): 795-801.
31. Cort WM. Antioxidant activity of tocopherols, ascorbyl palmitate, and ascorbic acid and their mode of action. J Am Oil Chem Soc. 1974; 51(7): 321-325.
32. Ross D, Mendiratta S, Qu Z- chao, Cobb CE, May JM. Ascorbate 6-palmitate protects human erythrocytes from oxidative damage. Free Radic Biol Med. 1999; 26(1): 81-89.
33. May JM, Qu Z-C, Cobb CE. Accessibility and reactivity of ascorbate 6-palmitate bound to erythrocyte membranes. Free Radic Biol Med. 1996; 21(4): 471-480.
34. Pokorski M, Marczak M, Dymecka A, Suchocki P. Ascorbyl palmitate as a carrier of ascorbate into neural tissues. J Biomed Sci. 2003; 10(2): 193-198.
35. Mäkinen M, Kähkönen M, Hopia A. Ascorbic acid and ascorbyl palmitate have only minor effect on the formation and decomposition of methyl linoleate hydroperoxides. Eur Jof Lipid Sci Technol. 2001; 103(10): 683-687.
36. Li Q, Wu Y, Chen XS, Zeng T, Liu LL, Feng ZQ, Liu DY, et al. Ascorbic acid 6-palmitate modulates microglia M1/M2 polarization in lipopolysaccharide-stimulated BV-2 cells via PERK/elF2α mediated endoplasmic reticulum stress. BMC Complement Med Ther. 2022 Nov 18;22(1):302.
37. Yilmaz O, Ozkan Y, Yildirim M, Oztürk AI, Erşan Y. Effects of alpha lipoic acid, ascorbic acid-6-palmitate, and fish oil on the glutathione, malonaldehyde, and fatty acids levels in erythrocytes of streptozotocin induced diabetic male rats. J Cell Biochem. 2002;86(3):530-9.
38. Bulama I, Saidu UF, Suleiman N, Abbas A, Saidu Y, Yakubu Y, et al. Antioxidative Strategy in Traumatic Brain Injury: Role of Low-Molecular-Weight Antioxidants. Exp Appl Med Sci. 2024; 4(4): 573-594.