Author(s):
Malarkodi K P, Sivaprasad R
Email(s):
pramalanand@gmail.com
DOI:
10.52711/0974-360X.2026.00154 
Address:
Malarkodi K P1,2*, Sivaprasad R1,2
1Department of Biochemistry, PSG College of Arts and Science, Civil Aerodrome Post, Coimbatore - 641014, Tamilnadu, India.
2Department of Medical Biochemistry, Dr. A. L. Mudaliar Post Graduate Institute of Basic Medical Sciences, University of Madras (Taramani), Chennai - 600113, Tamilnadu, India.
*Corresponding Author
Published In:
Volume - 19,
Issue - 3,
Year - 2026
ABSTRACT:
The use of adriamycin (ADR) as a potent chemotherapeutic agent is severely limited due to its cytotoxic side effects. The present study was carried out in Vero cell line to establish the protective role of lipoic acid (LA) in combating ADR-induced toxicities invitro. The highlight of the study was in assessing lipid peroxidation (LPO) and key cellular enzymes that were released in the spent medium as a result of ADR-induced cell damage. Apoptosis was assessed using the acridine orange and ethidium bromide (AO/EB) staining technique. Cells visualized under fluorescent microscopy showed yellow chromatin and orange chromatin indicating the different stages of apoptotic onset by ADR administration at two different time intervals. LPO and key cellular enzymes [lactate dehydrogenase (LDH), a cytoplasmic marker; N-acetyl-ß-glucosaminidase (NAG), a lysosomal marker and gamma glutamyl transferase (?-GT), a brush-border membrane marker] assessed showed a significant increase during ADR exposure. They were well under control upon pretreatment with LA. As a result, it can be deduced that a major factor contributing to ADR-induced apoptosis in Vero cells could be the result of reactive oxygen species (ROS) mediated oxidative stress that was opposed by LA pretreatment.
Cite this article:
Malarkodi K P, Sivaprasad R. Arbitration of Adriamycin - induced Apoptosis and Cellular Injury In Vero cells by DL-α-lipoic acid. Research Journal Pharmacy and Technology. 2026;19(3):1085-0. doi: 10.52711/0974-360X.2026.00154
Cite(Electronic):
Malarkodi K P, Sivaprasad R. Arbitration of Adriamycin - induced Apoptosis and Cellular Injury In Vero cells by DL-α-lipoic acid. Research Journal Pharmacy and Technology. 2026;19(3):1085-0. doi: 10.52711/0974-360X.2026.00154 Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2026-19-3-15
REFERENCES:
1. Galluzzi L, et al. Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death. Cell Death Differ. 2018; 25: 486–541. doi:10.1038/s41418-017-0012-4.
2. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007; 35 (4): 495–516. doi: 10.1080/01926230701320337.
3. Raudenská M, et al. Cell death in head and neck cancer pathogenesis and treatment. Cell Death Dis. 2021; 12: 192. doi: 10.1038/s41419-021-03474-5.
4. Podkowińska A, Formanowicz D. Chronic Kidney Disease as Oxidative Stress- and Inflammatory-Mediated Cardiovascular Disease. Antioxidants (Basel). 2020;9(8):752. doi: 10.3390/antiox9080752.
5. Archana V, Indumathy R. In- vitro Antioxidant and Cardio-protective effect of Delonix elata (L.) Leaf extract against Doxorubicin induced toxicity in H9c2 Cardio-myocyte cell line. Research J Pharm and Tech. 2021; 14(11): 5635-41. doi: 10.52711/0974-360X.2021.00980.
6. Satish Chandra A, Shanmugapandiyan P. Cardioprotective efficacy of Tridax procumbens methanolic extract in Doxorubicin induced Oxidative Cardiac Damage. Research J Pharm and Tech. 2020; 13(1): 110-13. doi: 10.5958/0974-360X.2020.00022.0
7. Doroshow JH. Mechanisms of anthracycline-enhanced reactive oxygen metabolism in tumor cells. Oxid Med Cell Longev. 2019; Dec 3. doi: 10.1155/2019/9474823.
8. Pourahmad J, et al. Role of oxygen free radicals in cancer development and treatment. Free Radic Dis 2016. Oct 26. doi.org/10.5772/64787.
9. Yang H, et al. The role of cellular reactive oxygen species in cancer chemotherapy. J Exp Clin Cancer Res. 2018; 37(1): 266. doi: 10.1186/s13046-018-0909-x.
10. Juan CA, et al. The Chemistry of Reactive Oxygen Species (ROS) Revisited: Outlining Their Role in Biological Macromolecules (DNA, Lipids and Proteins) and Induced Pathologies. Int J of Mol Sci. 2021; 22(9): 4642. doi: 10.3390/ijms22094642.
11. An X, et al. Oxidative cell death in cancer: mechanisms and therapeutic opportunities. Cell Death Dis. 2024; 15: 556. doi.org/10.1038/s41419-024-06939-5.
12. Chandrasekar R, et al. A Pharmacoeconomic Focus on Medicinal Plants with Anticancer Activity. Res J Pharmacognosy and Phytochem. 2018; 10(1): 91-100. doi: 10.5958/0975-4385.2018.00015.8.
13. Sritharan S, Sivalingam N. A comprehensive review on time-tested anticancer drug doxorubicin. Life Sci. 2021; 278: 119527. doi: 10.1016/j.lfs.2021.119527.
14. Taymaz-Nikerel H, et al. Doxorubicin induces an extensive transcriptional and metabolic rewiring in yeast cells. Sci Rep. 2018; 8(1): 13672. doi: 10.1038/s41598-018-31939-9.
15. Benzer F, et al. Chemoprotective Effects of curcumin on doxorubicin-induced nephrotoxicity in wistar rats: by modulating inflammatory cytokines, apoptosis, oxidative stress and oxidative DNA damage. Arch Physiol Biochem. 2018; 124(5): 448–57. doi: 10.1080/13813455.2017.1422766.
16. Aruna P, Gayathiri N M. Cardioprotective Activity of Telmisartan, Metformin and its Combination against Doxorubicin Induced Myocardial Infarction in Rat Model. Research J Pharm and Tech. 2018; 11(12): 5293-96. doi: 10.5958/0974-360X.2018.00964.2.
17. Ahmed AZ, et al. Methyl Gallate Mitigates Doxorubicin-Induced Peripheral Cytopenias: A Preclinical Experimental Study. Research J Pharm and Tech. 2021; 14(9): 4529-4. doi: 10.52711/0974-360X.2021.00788.
18. Jalaiah M, et al. Krill Oil attenuates doxorubicin induced cognitive impairment by inhibiting oxidative stress markers in the brains of wistar rats. Research J Pharm and Tech. 2023; 16(2): 652-8. doi: 10.52711/0974-360X.2023.00111.
19. Mani V, et al. Levetiracetam mitigates doxorubicin-induced Oxidative stress associated with Cognitive impairments and Neurodegeneration in experimental rats: In vivo and Molecular Modelling Studies. Research J Pharm and Tech. 2024; 17(7): 3261-7. doi: 10.52711/0974-360X.2024.00511.
20. Sivaprasad R, et al. Combined effects of lipoic acid and dimercaptosuccinic acid in improving lead-induced alterations on glucose metabolism, energy metabolism and tissue histology. Ind J Clin Biochem. 2025. doi.org/10.1007/s12291-025-01330-3.
21. Kushwaha K, et al. Epigenetic modifying potential of Lipoic acid: Implications in curing diabetes. Research J Pharm and Tech. 2021; 14(12): 6747-2. doi: 10.52711/0974-360X.2021.01165.
22. Vani R, et al. Analytical Method Development and Validation for the Determination of Allopurinol and Alphalipoic Acid Using Reverse Phase HPLC Method in Bulk and Tablet Dosage Form. Research J Pharm and Tech. 2015; 8(2): 207-11. doi: 10.5958/0974-360X.2015.00038.4.
23. Chen Y, et al. Research progress on morphology and mechanism of programmed cell death. Cell Death. 2024; (15): 327. doi.org/10.1038/s41419-024-06712-8.
24. Gherghi IC, et al. Study of interactions between DNA-ethidium bromide (EB) and DNA-acridine orange (AO), in solution, using hanging mercury drop electrode (HMDE) Talanta. 2003; 61: 103–12. doi: 10.1016/S0039-9140(03)00238-8.
25. Baskić D, et al. Analysis of cycloheximide-induced apoptosis in human leukocytes: fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide. Cell Biol Int. 2006; 30: 924–32. doi: 10.1016/j.cellbi.2006.06.016.
26. Renvoize C, et al. Apoptosis: identification of dying cells. Cell Biol Toxicol. 1998; 14(2): 111-20. doi: 10.1023/a:1007429904664.
27. Krishnasamy L, et al. Anticancer property of Colchicine isolated from Indigofera aspalathoids. Research J Pharm and Tech. 2016; 9(4): 386-90. doi: 10.5958/0974-360X.2016.00069. X.
28. Devasagayam TPA, et al. Activity of thiols as singlet molecular quenchers. J Photochem Photobiol B. 1999; 9 (1): 105-16. doi: 10.1016/1011-1344(91)80008-6.
29. King J. The dehydrogenases or oxidoneductase-lactate dehydrogenase. In: Practical clinical enzymology. Van, D. (ed.), Nostrand Company Ltd., London, 1965, pp.83-93.
30. Maruhn D. Rapid colorimetric assay of β-galactosidase and N-acetyl-β-D-galactosaminidase in human urine. Clin Chim Acta. 1976; 73 (3): 453-61. doi: 10.1016/0009-8981(76)90147-9.
31. Orlowski M, Meister A. Isolation of γ-glutamyl transpeptidase from hog kidney. J Biol Chem. 1965; 240(1): 338-347. doi.org/10.1016/S0021-9258(18)97654-4.
32. Anggi V, Adikusuma W. Total Antioxidant and In-Vitro Cytotoxic of Abelmoschus Manihot (L.) Medik from palu of Central Sulawesi and Doxorubicin on 4T1 cells line and Vero Cells. Research J Pharm and Tech. 2019; 12(11): 5472-76. doi: 10.5958/0974-360X.2019.00949.1
33. Divya B, et al. Cardioprotective activity of Lawsonia inermis roots against Doxorubicin Treated Mice. Research J Pharm and Tech. 2020; 13(7): 3279-3283. doi: 10.5958/0974-360X.2020.00581.8
34. Baxter-Holland M, Dass CR. Doxorubicin, mesenchymal stem cell toxicity and antitumour activity: implications for clinical use. J Pharm Pharmacol. 2018; 70(3): 320-27. doi: 10.1111/jphp.12869.
35. Gilleron et al. NADPH oxidases participate to doxorubicin-induced cardiac myocyte apoptosis. Biochem Biophys Res Commun. 2009; 388(4): 727-31. doi: 10.1016/j.bbrc.2009.08.085.
36. Verhaegen S, et al. Inhibition of apoptosis by antioxidants in the human HL-60 leukemia cell line. Biochem Pharmacol. 1995; 50(7): 1021-29. doi: 10.1016/0006-2952(95)00233-p.
37. Handelman GJ, et al. Alpha-lipoic acid reduction by mamalian cells to the dithiol form and release into the culture medium. Biochem Pharmacol. 1994; 47(10): 1725-30. doi: 10.1016/0006-2952(94)90298-4.
38. Pierce RH, et al. Disruption of redox homeostasis in tumour necrosis factor-induced apoptosis in a murine hepatocyte cell line. Am J Pathol. 2000; 157(1): 221-36. doi: 10.1016/S0002-9440(10)64533-6.
39. Sen CK, et al. Fas mediated apoptosis of human Jurkat T-cells: Intracellular events and potentiation by redox-active alpha-lipoic acid. Cell Death Differ. 1999; 6(5): 481-91. doi: 10.1038/sj.cdd.4400514.
40. Ritov VB, et al. Non-random peroxidation of different classes of membrane phospholipids in live cells detected by metabolically integrated cis- parinaric acid. Biochim Biophys Acta. 1996; 1283(2): 127-40. doi.org/10.1016/0005-2736(96)00083-1
41. Benchekroun MN, Robert J. Measurement of doxorubicin-induced lipid peroxidation under the conditions that determine cytotoxicity in cultured tumor cells. Anal Biochem. 1992 201(2): 326-30. doi: 10.1016/0003-2697(92)90346-9.
42. Zhao S, et al. The RNA binding protein hnRNPK protects against adriamycin-induced podocyte injury. Ann Transl Med. 2021; 9(16):1303. doi: 10.21037/atm-21-3577.
43. Hanigan MH, Frierson HF Jr. Immunohistochemical detection of gamma-glutamyl transpeptidase in normal human tissue. J Histochem Cytochem. 1996; 44(10):1101-8. doi: 10.1177/44.10.8813074.
44. Sivaprasad TR, et al. Therapeutic efficacy of lipoic acid in combination with dimercaptosuccinic acid against lead-induced renal tubular defects and on isolated kidney brush-border enzyme activities. Chem Biol Interact. 2004; 147(3): 259-271. doi: 10.1016/j.cbi.2004.01.004.