EVALUATION OF SERUM LEVELS OF CYP2D6, CYP3A4, GSH, AND MDA AS PREDICTIVE BIOMARKERS FOR METHAMPHETAMINE-INDUCED LIVER DYSFUNCTION USING ADVANCED STATISTICAL MODELING
DOI:
https://doi.org/10.22159/ijap.2026v18i2.57318Keywords:
Methamphetamine, Cytochrome P-450 CYP2D6, Cytochrome P-450 CYP3A4, Glutathione, Malondialdehyde, Principal component analysisAbstract
Objective: This study aimed to evaluate the predictive potential of serum protein levels of CYP2D6, CYP3A4, glutathione (GSH), and malondialdehyde (MDA) as early biomarkers of METH-induced liver dysfunction using multivariate statistical approaches, compared to standard LFTs.
Methods: This case–control study included 90 METH users and 45 healthy control individuals. Serum CYP2D6, CYP3A4, GSH, and MDA levels, and routine LFTs were determined. Analysis was performed using principal component analysis (PCA), receiver operating characteristic (ROC) curves, and decision tree modeling.
Results: METH users showed significantly decreased serum levels of CYP2D6, CYP3A4, and GSH and higher MDA levels (p<0.0001), suggestive of oxidative metabolic disequilibrium. While most routine LFTs were normal, other markers, including AST, AST/ALT ratio, and albumin-to-globulin ratio, increased significantly. PCA demonstrated that early injury components could be divided into separate groups according to the markers of their metabolic and oxidative components. CYP2D6 ≤ 1.98 ng/ml was the best discriminating predictor according to the decision tree with 93.3% accuracy (AUC = 94).
Conclusion: Serum protein levels of CYP2D6, CYP3A4, GSH, and MDA demonstrated high sensitivity for detecting subclinical liver injury in METH users before conventional LFTs became abnormal. Their integration into diagnostic models may facilitate early interventions in high-risk populations.
References
1. Kondza M, Brizic I, Jokic S. Flavonoids as CYP3A4 inhibitors in vitro. Biomedicines. 2024 Mar;12(3):644. doi: 10.3390/biomedicines12030644, PMID 38540257.
2. Elfaki I, Almutairi FM, Mir R, Khan R, Abu-duhier F. Cytochrome P450 CYP1B12 gene and its association with T2D in tabuk population, northwestern region of Saudi Arabia. Asian J Pharm Clin Res. 2018 Jan 1;11(1):55-9. doi: 10.22159/ajpcr.2018.v11i1.21657.
3. Zhao M, Ma J, Li M, Zhang Y, Jiang B, Zhao X. Cytochrome P450 enzymes and drug metabolism in humans. Int J Mol Sci. 2021 Nov;22(23):12808. doi: 10.3390/ijms222312808, PMID 34884615.
4. Kumar A, Estrada DF. Structural basis of bidirectional allostery across the heme in a cytochrome P450 enzyme. J Biol Chem. 2023;299(8):104977. doi: 10.1016/j.jbc.2023.104977, PMID 37390989.
5. Loos NH, Beijnen JH, Schinkel AH. The mechanism-based inactivation of CYP3A4 by ritonavir: what mechanism? Int J Mol Sci. 2022 Aug;23(17):9866. doi: 10.3390/ijms23179866, PMID 36077262.
6. Rohmanika R, Arozal W, Louisa M, Gayatri A, Kekalih A, Marzuki HY JE. Patterns of methamphetamine use and its associations with psychiatric symptoms in patients upon admission at the national rehabilitation center lido bogor Indonesia. Int J Appl Pharm. 2022;14(5):96-100. doi: 10.22159/ijap.2022.v14s5.16.
7. Frederiksen T. Using population pharmacokinetic analyses of drugs metabolized by CYP2D6 to study the genotype-phenotype translation. Basic Clin Pharmacol Toxicol. 2023 Aug;133(2):113-23. doi: 10.1111/bcpt.13903, PMID 37221697.
8. Tu DZ, Hu XY, Lei JX, Liu SY, Xiao ZP, Yang L. A patent review of CYP3A4 inhibitors (2018-present). Expert Opin Ther Pat. 2025 May;35(5):503-13. doi: 10.1080/13543776.2025.2470294, PMID 39976548.
9. Yoon S, Jeong S, Jung E, Kim KS, Jeon I, Lee Y. Effect of CYP3A4 metabolism on sex differences in the pharmacokinetics and pharmacodynamics of zolpidem. Sci Rep. 2021 Sep;11(1):19150. doi: 10.1038/s41598-021-98689-z, PMID 34580385.
10. Ma Y, Wu H, Wang H, Chen F, Xie Z, Zhang Z. Psychiatric comorbidities and liver injury are associated with unbalanced plasma bile acid profile during methamphetamine withdrawal. Front Endocrinol (Lausanne). 2021;12:801686. doi: 10.3389/fendo.2021.801686, PMID 35046900.
11. Lewis D, Kenneally M, Van Denheuvel C, Byard RW. Methamphetamine deaths: changing trends and diagnostic issues. Med Sci Law. 2021 Apr;61(2):130-7. doi: 10.1177/0025802420986707, PMID 33423599.
12. Al-Hakeim HK, Altufaili MF, Almulla AF, Moustafa SR, Maes M. Increased lipid peroxidation and lowered antioxidant defenses predict methamphetamine induced psychosis. Cells. 2022 Nov;11(22):3694. doi: 10.3390/cells11223694, PMID 36429122.
13. Ramli FF, Rejeki PS, Ibrahim N, Abdullayeva G, Halim S. A mechanistic review on toxicity effects of methamphetamine. Int J Med Sci. 2025;22(3):482-507. doi: 10.7150/ijms.99159, PMID 39898237.
14. Song YY, Lu Y. Decision tree methods: applications for classification and prediction. Shanghai Arch Psychiatry. 2015 Apr;27(2):130-5. doi: 10.11919/j.issn.1002-0829.215044, PMID 26120265.
15. Dida N, Kassa Y, Sirak T, Zerga E, Dessalegn T. Substance use and associated factors among preparatory school students in bale zone oromia regional state southeast Ethiopia. Harm Reduct J. 2014 Aug;11(21):21. doi: 10.1186/1477-7517-11-21, PMID 25108629.
16. Jolliffe IT, Cadima J. Principal component analysis: a review and recent developments. Philos Trans A Math Phys Eng Sci. 2016 Apr;374(2065):20150202. doi: 10.1098/rsta.2015.0202, PMID 26953178.
17. Dostalek M, Jurica J, Pistovcakova J, Hanesova M, Tomandl J, Linhart I. Effect of methamphetamine on cytochrome P450 activity. Xenobiotica. 2007 Dec;37(12):1355-66. doi: 10.1080/00498250701652877, PMID 17922362.
18. Yan B, Ye X, Wang J, Han J, Wu L, He S. An algorithm framework for drug-induced liver injury prediction based on genetic algorithm and ensemble learning. Molecules. 2022 May;27(10):3112. doi: 10.3390/molecules27103112, PMID 35630587.
19. Zhang KK, Wang H, Qu D, Chen LJ, Wang LB, Li JH. Luteolin alleviates methamphetamine-induced hepatotoxicity by suppressing the p53 pathway-mediated apoptosis, autophagy and inflammation in rats. Front Pharmacol. 2021;12:641917. doi: 10.3389/fphar.2021.641917, PMID 33679421.
20. Rifai N. Tietz textbook of clinical chemistry and molecular diagnostics. 7th ed. St. Louis, MO: Elsevier Health Sciences; 2017. Available from: https://www.nursingbridgetestbank.Com/wpcontent/uploads/2025/12/Download-Sample.pdf.
21. Elliott AC, Woodward WA. Statistical analysis quick reference guidebook: with SPSS examples. Sage Publications; 2007.
22. Fu S, Wu D, Jiang W, Li J, Long J, Jia C. Molecular biomarkers in drug-induced liver injury: challenges and future perspectives. Front Pharmacol. 2020;10:1667. doi: 10.3389/fphar.2019.01667, PMID 32082163.
23. Gerth K, Kodidela S, Mahon M, Haque S, Verma N, Kumar S. Circulating extracellular vesicles containing xenobiotic metabolizing CYP enzymes and their potential roles in extrahepatic cells via cell–cell interactions. Int J Mol Sci. 2019;20(24):6178. doi: 10.3390/ijms20246178, PMID 31817878.
24. Palomo L, Mleczko JE, Azkargorta M, Conde Vancells J, Gonzalez E, Elortza F. Abundance of cytochromes in hepatic extracellular vesicles is altered by drugs related with drug-induced liver injury. Hepatol Commun. 2018;2(9):1064-79. doi: 10.1002/hep4.1210, PMID 30202821.
25. Soo JY, Wiese MD, Dyson RM, Gray CL, Clarkson AN, Morrison JL. Methamphetamine administration increases hepatic CYP1A2 but not CYP3A activity in female guinea pigs. PLOS One. 2020;15(5):e0233010. doi: 10.1371/journal.pone.0233010, PMID 32396581.
26. Loftis JM, Janowsky A. Neuroimmune basis of methamphetamine toxicity. Int Rev Neurobiol. 2014;118:165-97. doi: 10.1016/B978-0-12-801284-0.00007-5, PMID 25175865.
27. Wang L, Bai M, Jin T, Zheng J, Wang Y, He Y. Effects of CYP3A4 polymorphisms on drug addiction risk among the Chinese han population. Front Public Health. 2019;7:315. doi: 10.3389/fpubh.2019.00315, PMID 31799230.
28. Zhou S, Cheng K, Peng Y, Liu Y, Hu Q, Zeng S. Regulation mechanism of endoplasmic reticulum stress on metabolic enzymes in liver diseases. Pharmacol Res. 2024 Sep;207:107332. doi: 10.1016/j.phrs.2024.107332, PMID 39089398.
29. Harbrecht BG, Frye RF, Zenati MS, Branch RA, Peitzman AB. Cytochrome P-450 activity is differentially altered in severely injured patients. Crit Care Med. 2005;33(3):541-6. doi: 10.1097/01.CCM.0000155989.54344.E0, PMID 15753745.
30. Hossam Abdelmonem B, Abdelaal NM, Anwer EK, Rashwan AA, Hussein MA, Ahmed YF. Decoding the role of CYP450 enzymes in metabolism and disease: a comprehensive review. Biomedicines. 2024 Jul;12(7):1467. doi: 10.3390/biomedicines12071467, PMID 39062040.
31. McDonnell-Dowling K, Kelly JP. The role of oxidative stress in methamphetamine induced toxicity and sources of variation in the design of animal studies. Curr Neuropharmacol. 2017;15(2):300-14. doi: 10.2174/1570159x14666160428110329, PMID 27121285.
32. Yamamoto BK, Moszczynska A, Gudelsky GA. Amphetamine toxicities: classical and emerging mechanisms. Ann N Y Acad Sci. 2010 Feb;1187:101-21. doi: 10.1111/j.1749-6632.2009.05141.x, PMID 20201848.
33. Turan C, Senormancı G, Neselioglu S, Budak Y, Erel O, Senormancı O. Oxidative stress and inflammatory biomarkers in people with methamphetamine use disorder. Clin Psychopharmacol Neurosci. 2023 Aug;21(3):572-82. doi: 10.9758/cpn.22.1047, PMID 37424424.
34. Bogoje Raspopovic A, Balta V, Vodopic M, Drobac M, Boros A, Dikic D. The possible role of oxidative stress marker glutathione in the assessment of cognitive impairment in multiple sclerosis. Open Med (Wars). 2024;19(1):20240952. doi: 10.1515/med-2024-0952, PMID 38623459.
35. Cherian DA, Peter T, Narayanan A, Madhavan SS, Achammada S, Vynat GP. Malondialdehyde as a marker of oxidative stress in periodontitis patients. J Pharm Bioallied Sci. 2019 May;11(Suppl 2):S297-300. doi: 10.4103/JPBS.JPBS_17_19, PMID 31198357.
36. Eskandari MR, Rahmati M, Khajeamiri AR, Kobarfard F, Noubarani M, Heidari H. A new approach on methamphetamine-induced hepatotoxicity: involvement of mitochondrial dysfunction. Xenobiotica. 2014 Jan;44(1):70-6. doi: 10.3109/00498254.2013.807958, PMID 23786375.
37. Ajoolabady A, Kaplowitz N, Lebeaupin C, Kroemer G, Kaufman RJ, Malhi H. Endoplasmic reticulum stress in liver diseases. Hepatology. 2023 Feb;77(2):619-39. doi: 10.1002/hep.32562, PMID 35524448.
38. Mohammed NS, Ali ZQ, Mohamed AS, Mirza SA. The impact of methamphetamine on liver injury in Iraqi male addicts. Toxicol Rep. 2024 Dec;13:101806. doi: 10.1016/j.toxrep.2024.101806, PMID 39624223.
39. Bihari S, Bannard Smith J, Bellomo R. Albumin as a drug: its biological effects beyond volume expansion. Crit Care Resusc. 2020 Sep;22(3):257-65. doi: 10.1016/S1441-2772(23)00394-0, PMID 32900333.
40. Damborska A, Hanakova L, Pindurova E, Horska K. Case report: therapeutic drug monitoring and CYP2D6 phenoconversion in a protracted paroxetine intoxication. Front Pharmacol. 2024;15:1444857. doi: 10.3389/fphar.2024.1444857, PMID 39295933.
41. Zhou Z, Chen C, Sun M, Xu X, Liu Y, Liu Q. A decision tree model to predict liver cirrhosis in hepatocellular carcinoma patients: a retrospective study. Peer J. 2023;11:e15950. doi: 10.7717/peerj.15950, PMID 37641600.
42. Ouyang X, Fan Q, Ling G, Shi Y, Hu F. Identification of diagnostic biomarkers and subtypes of liver hepatocellular carcinoma by multi-omics data analysis. Genes (Basel). 2020 Sep;11(9):1051. doi: 10.3390/genes11091051, PMID 32899915.
43. Kumar S, Sinha N, Gerth KA, Rahman MA, Yallapu MM, Midde NM. Specific packaging and circulation of cytochromes P450, especially 2E1 isozyme in human plasma exosomes and their implications in cellular communications. Biochem Biophys Res Commun. ScienceDirect. 2017;491(3):675-80. doi: 10.1016/j.bbrc.2017.07.145, PMID 28756226.
44. Kumar S, Singla B, Singh AK, Thomas Gooch SM, Zhi K, Singh UP. Hepatic extrahepatic and extracellular vesicle cytochrome P450 2E1 in alcohol and acetaminophen-mediated adverse interactions and potential treatment options. Cells. 2022;11(17):2620. doi: 10.3390/cells11172620, PMID 36078027.
Published
How to Cite
Issue
Section
Copyright (c) 2026 ALI FARIED SALMAN, GHID HASSAN ABDULHADI, MUSHTAQ TALIB HASHIM
This work is licensed under a Creative Commons Attribution 4.0 International License.