Protective effects of red ginger extract (RG) on oxidative stress markers in a zebrafish model of gestational diabetes mellitus
Abstract
Background: Gestational diabetes mellitus (GDM) results from pregnancy-induced insulin resistance and increases oxidative stress, marked by decreased Superoxide Dismutase (SOD) and elevated Malondialdehyde (MDA). Current therapies, including Insulin and Metformin, have limitations during pregnancy. Red ginger extract (RG) (Zingiber officinale var. rubrum) contains antioxidant compounds and may reduce oxidative stress.
Objective: This study aimed to analyze the effect of red ginger extract (RG) on SOD and MDA levels in zebrafish larvae as hyperglycemia-induced oxidative stress relevant to GDM.
Methods: This study used a true experimental laboratory design with a post-test only control group approach conducted from January to April 2026 at the Pharmacology Laboratory. A total of 450 zebrafish embryos were allocated into five groups consisting of three biological replicates of embryos each (90 embryos/group): a control group, 3% glucose, and three treatment groups receiving red ginger extract (RG) at 5, 10, and 15 µg/mL. Hyperglycemia was induced from 2–72 hpf, and SOD and MDA levels were measured at 6 dpf using ELISA.
Results: The 3% glucose group exhibited reduced SOD and increased MDA levels compared with controls. Red ginger extract (RG)at all doses tended to increase SOD levels, although the differences were not statistically significant. MDA levels were significantly reduced in all treatment groups, with the most optimal effect observed at a dose of 10 µg/mL. SOD and MDA levels showed a significant negative correlation with moderate strength.
Conclusion: Red ginger extract (RG) may serve as a potential complementary antioxidant agent.
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References
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3. Adli FK. Diabetes Melitus Gestasional : Diagnosis dan Faktor Risiko. Jurnal Medika Hutama. 2021;03(01):1545-1551.
4. González P, Lozano P, Ros G, Solano F. Hyperglycemia and Oxidative Stress: An Integral, Updated and Critical Overview of Their Metabolic Interconnections. Int J Mol Sci. 2023;24(11). doi:10.3390/ijms24119352
5. Zheng ZX, Feng X, Zhuang L. The Effect of Oxidative Stress and Antioxidants Treatment on Gestational Diabetes Mellitus Outcome: A Scoping Review. Cell Biochem Biophys. Springer. 2024;82(4):3003-3013. doi:10.1007/s12013-024-01417-3
6. Mittal R, Prasad K, Lemos JRN, Arevalo G, Hirani K. Unveiling Gestational Diabetes: An Overview of Pathophysiology and Management. Int J Mol Sci. 2025;26(5). doi:10.3390/ijms26052320
7. Naureen T, Syeda Fouzia Mazloum, Laiba Shaukat, Qural UL ain Fatima, Sadia Mubarak, Mahvash Khan. SERUM MALONDIALDEHYDE AS A MARKER OF OXIDATIVE STRESS IN GESTATIONAL DIABETES MELLITUS: A COMPARATIVE STUDY. Pakistan Journal of Physiology. 2025;21(3):3-6. doi:10.69656/pjp.v21i3.1836
8. Asfandiyar, Hadi N, Ali Zaidi I, et al. Estimation of Serum Malondialdehyde (a Marker of Oxidative Stress) as a Predictive Biomarker for the Severity of Coronary Artery Disease (CAD) and Cardiovascular Outcomes. Cureus. Published online September 20, 2024. doi:10.7759/cureus.69756
9. Plows JF, Stanley JL, Baker PN, Reynolds CM, Vickers MH. The pathophysiology of gestational diabetes mellitus. Int J Mol Sci. 2018;19(11). doi:10.3390/ijms19113342
10. Mirabelli M, Tocci V, Donnici A, et al. Maternal Preconception Body Mass Index Overtakes Age as a Risk Factor for Gestational Diabetes Mellitus. J Clin Med. 2023;12(8). doi:10.3390/jcm12082830
11. Fernández-Valero A, Peña-Montero N, Lima-Rubio F, et al. Changes in Oxidative Stress and Intestinal Permeability during Pregnancy in Women with Gestational Diabetes Mellitus Treated with Metformin or Insulin and Healthy Controls: A Randomized Controlled Trial. Antioxidants. 2023;12(11). doi:10.3390/antiox12111981
12. Hajimoosayi F, Sadatmahalleh SJ, Kazemnejad A, Pirjani R. Effect of ginger on the blood glucose level of women with gestational diabetes mellitus (GDM) with impaired glucose tolerance test (GTT): A randomized doubleblind placebo-controlled trial. BMC Complement Med Ther. 2020;20(1). doi:10.1186/s12906-020-02908-5
13. Paudel KR, Orent J, Penela OG. Pharmacological properties of ginger (Zingiber officinale): what do meta-analyses say? a systematic review. Front Pharmacol. Frontiers Media SA. 2025;16. doi:10.3389/fphar.2025.1619655
14. Ayustaningwarno F, Anjani G, Ayu AM, Fogliano V. A critical review of Ginger’s (Zingiber officinale) antioxidant, anti-inflammatory, and immunomodulatory activities. Front Nutr. 2024;11. doi:10.3389/fnut.2024.1364836
15. Manjunathan T, Guru A, Arokiaraj J, Gopinath P. 6-Gingerol and Semisynthetic 6-Gingerdione Counteract Oxidative Stress Induced by ROS in Zebrafish. Chem Biodivers. 2021;18(12). doi:10.1002/cbdv.202100650
16. Ayustaningwarno F, Anjani G, Ayu AM, Fogliano V. A critical review of Ginger’s (Zingiber officinale) antioxidant, anti-inflammatory, and immunomodulatory activities. Front Nutr. 2024;11. doi:10.3389/fnut.2024.1364836
17. Yang L, Yang F, Teng L, Katayama I. 6-shogaol protects human melanocytes against oxidative stress through activation of the nrf2-antioxidant response element signaling pathway. Int J Mol Sci. 2020;21(10). doi:10.3390/ijms21103537
18. Heckler K, Kroll J. Zebrafish as a model for the study of microvascular complications of diabetes and their mechanisms. Int J Mol Sci. 2017;18(9). doi:10.3390/ijms18092002
19. Alla RK, Konakanchi A, Nallakunta R, A. V. R, M. C. SS, Mantena SR. The Zebrafish (Danio rerio): A Dynamic Model Organism for Scientific Research. UTTAR PRADESH JOURNAL OF ZOOLOGY. 2024;45(15):526-540. doi:10.56557/upjoz/2024/v45i154269
20. Khotimah H, Prima WE, Weningtyas A, et al. Neuroprotective Activity and Antioxidant Effect of Salacca zalacca Peel Ethanol Extract on High Glucose Induced Zebrafish (Danio rerio) Embryo. Tropical Journal of Natural Product Research. 2021;5(12):2079-2084. doi:10.26538/tjnpr/v5i12.6
21. Konadu B, Cox CK, Garrett MR, Gibert Y. Excess glucose or fat differentially affects metabolism and appetite-related gene expression during zebrafish embryogenesis. iScience. 2023;26(7). doi:10.1016/j.isci.2023.107063
22. Darwitri, Yuliyani T, Nuraenah E, et al. Centella asiatica increased the body length through the modulation of antioxidant in rotenone-induced zebrafish larvae. Biomedical and Pharmacology Journal. 2018;11(2):827-833. doi:10.13005/bpj/1438
23. Thompson E, Hensley J, Taylor RS. Effect of High Glucose on Embryological Development of Zebrafish, Brachyodanio rerio through Wnt Pathway. Published online July 11, 2024. doi:10.20944/preprints202407.0951.v1
24. Saucedo R, Ortega-Camarillo C, Ferreira-Hermosillo A, Díaz-Velázquez MF, Meixueiro-Calderón C, Valencia-Ortega J. Role of Oxidative Stress and Inflammation in Gestational Diabetes Mellitus. Antioxidants. 2023;12(10). doi:10.3390/antiox12101812
25. Ibrahim A, Khoo MI, Ismail EHE, et al. Oxidative stress biomarkers in pregnancy: a systematic review. Reproductive Biology and Endocrinology. 2024;22(1). doi:10.1186/s12958-024-01259-x
26. Luo L, Zhu S, Akbari A, Tan B. Ginger Could Improve Gestational Diabetes by Targeting Genes Involved in Nutrient Metabolism, Oxidative Stress, Inflammation, and the WNT/β-Catenin/GSK3β Signaling Pathway. Nat Prod Commun. 2022;17(12). doi:10.1177/1934578X221141276
27. Zhang S, Kou X, Zhao H, Mak KK, Balijepalli MK, Pichika MR. Zingiber officinale var. rubrum: Red Ginger’s Medicinal Uses. Molecules. 2022;27(3). doi:10.3390/molecules27030775
28. Handayani, Serihati AYT, Yuniarti CA, et al. Metode Penelitian Bagi Mahasiswa Dan Tenaga Kesehatan. (Arsya UT, ed.). PT Adab Indonesia; 2025. https://www.google.co.id/books/edition/Metode_Penelitian_Bagi_Mahasiswa_Dan_Ten/no1gEQAAQBAJ?hl=en&gbpv=0
29. Wang R, Liu K, Zhang Y, Chen X, Wang X. Evaluation of the Developmental Toxicity Induced by E804 in Zebrafish Embryos. Front Pharmacol. 2020;11. doi:10.3389/fphar.2020.00032
30. OECD/OCDE. Test Guideline No. 236 Fish Embryo Acute Toxicity (FET) Test. In: OECD Guidelines for the Testing of Chemicals. Section 2. Effects on Biotic Systems; 2025. http://www.oecd.org/termsandconditions/
31. Khotimah H, Prima WE, Weningtyas A, et al. Neuroprotective Activity and Antioxidant Effect of Salacca zalacca Peel Ethanol Extract on High Glucose Induced Zebrafish (Danio rerio) Embryo. Tropical Journal of Natural Product Research. 2022;5(12):2079-2084. doi:10.26538/tjnpr/v5i12.6
32. Ibrahim AH, Hasan H, Sy. Pakaya M. SKRINING FITOKIMIA DAN UJI DAYA HAMBAT EKTRAK DAUN JAHE MERAH (Zingiber officinale var rubrum) TERHADAP BAKTERI Staphylococcus Epidermidis DAN Escherichia Coli. Indonesian Journal of Pharmaceutical Education. 2021;1(2):107-118. doi:10.37311/ijpe.v1i2.10547
33. Hoyberghs J, Bars C, Ayuso M, Van Ginneken C, Foubert K, Van Cruchten S. DMSO Concentrations up to 1% are Safe to be Used in the Zebrafish Embryo Developmental Toxicity Assay. Frontiers in Toxicology. 2021;3(804033). doi:10.3389/ftox.2021.804033
34. Nam YH, Hong BN, Rodriguez I, et al. Steamed ginger may enhance insulin secretion through Katp channel closure in pancreatic β-cells potentially by increasing 1-dehydro-6-gingerdione content. Nutrients. 2020;12(2):1-15. doi:10.3390/nu12020324
35. Zhang S, Kou X, Zhao H, Mak KK, Balijepalli MK, Pichika MR. Zingiber officinale var. rubrum: Red Ginger’s Medicinal Uses. Molecules. 2022;27(3). doi:10.3390/molecules27030775
36. Ahmadifar E, Sheikhzadeh N, Roshanaei K, Dargahi N, Faggio C. Can dietary ginger (Zingiber officinale)alter biochemical and immunological parameters and gene expression related to growth, immunity and antioxidant system in zebrafish (Danio rerio)? Aquaculture. 2019;507:341-348. doi:10.1016/j.aquaculture.2019.04.049
37. Matin M, Wysocki K, Horbańczuk JO, Rossi L, Atanasov AG. Ginger (Zingiber officinale) dietary supplementation in mice regulates liver antioxidant defense systems in a dose- and age-dependent. Front Pharmacol. 2025;16. doi:10.3389/fphar.2025.1597599
38. Darwitri, Yuliyani T, Nuraenah E, et al. Centella asiatica increased the body length through the modulation of antioxidant in rotenone-induced zebrafish larvae. Biomedical and Pharmacology Journal. 2018;11(2):827-833. doi:10.13005/bpj/1438
39. Hajimoosayi F, Sadatmahalleh SJ, Kazemnejad A, Pirjani R. Effect of ginger on the blood glucose level of women with gestational diabetes mellitus (GDM) with impaired glucose tolerance test (GTT): A randomized doubleblind placebo-controlled trial. BMC Complement Med Ther. 2020;20(1). doi:10.1186/s12906-020-02908-5
40. Tumilaar SG, Hardianto A, Dohi H, Kurnia D. A Comprehensive Review of Free Radicals, Oxidative Stress, and Antioxidants: Overview, Clinical Applications, Global Perspectives, Future Directions, and Mechanisms of Antioxidant Activity of Flavonoid Compounds. J Chem. Hindawi Limited. 2023;2024. doi:10.1155/2024/5594386
41. Rossi M, Teitsworth TS, McKenzie E, Caruso A, Thieke N, Caruso F. Ginger (Zingiber officinale) and Zingerone Antioxidant Properties Studied Using Hydrodynamic Voltammetry, Zingerone Crystal Structure and Density Functional Theory (DFT)—Results Support Zingerone Experimental Catalytic Behavior Similar to Superoxide Dismutases (SODs). Int J Mol Sci. 2025;26(21). doi:10.3390/ijms262110645
42. Fernández-Valero A, Peña-Montero N, Lima-Rubio F, et al. Changes in Oxidative Stress and Intestinal Permeability during Pregnancy in Women with Gestational Diabetes Mellitus Treated with Metformin or Insulin and Healthy Controls: A Randomized Controlled Trial. Antioxidants. 2023;12(11). doi:10.3390/antiox12111981
43. Saucedo R, Ortega-Camarillo C, Ferreira-Hermosillo A, Díaz-Velázquez MF, Meixueiro-Calderón C, Valencia-Ortega J. Role of Oxidative Stress and Inflammation in Gestational Diabetes Mellitus. Antioxidants. 2023;12(1812). doi:10.3390/antiox12101812
44. Sun C, Shen J, Fang R, et al. The impact of environmental and dietary exposure on gestational diabetes mellitus: a comprehensive review emphasizing the role of oxidative stress. Front Endocrinol (Lausanne). 2025;16(1393883). doi:10.3389/fendo.2025.1393883
45. Paudel KR, Orent J, Penela OG. Pharmacological properties of ginger (Zingiber officinale): what do meta-analyses say? a systematic review. Front Pharmacol. 2025;16. doi:10.3389/fphar.2025.1619655
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Authors
Purwasuyitno, D. . N., Astuti, C. N. K., Florencia, D. J., Rahardjo, B., & Sulistomo, H. W. (2026). Protective effects of red ginger extract (RG) on oxidative stress markers in a zebrafish model of gestational diabetes mellitus. JURNAL RISET KESEHATAN POLTEKKES DEPKES BANDUNG, 18(2), 520–535. https://doi.org/10.34011/juriskesbdg.v18i2.3322
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