Virgin Coconut Oil and Folic Acid Improve Insulin Signaling and Cardiac Function in Rotenone Induced Stunted Zebrafish Larvae

Syahana Aini, Farica Emiliana, Hanida Aisyah Ardiana, Nurdiana Nurdiana, Ariani Ariani, Brigitta Ida Resita Vebrianti Corebima

Abstract


Stunting is a chronic growth disorder caused by prolonged nutritional deficiencies and environmental stress, resulting in reduced height for age. Mitochondrial dysfunction and oxidative stress are increasingly recognized as key contributors to impaired growth and cardiometabolic instability during early development. This study investigated the potential protective effects of Virgin Coconut Oil (VCO) and folic acid on insulin signaling and cardiac finction in a rotenone induced stunted zebrafish larval model. A laboratory experimental study with a true experimental post test only controlled group design was conducted among 30 Zebrafish (Danio rerio) for Irs gene expression at 9 day post fertilization (dpf) as samples per tube, heart rate was measured at 3, 6, and 9 dpf. The total group treatment was five with inclusion and exclusion criterias, negative control (NC), positive control exposed to rotenone (12.5 ppb), VCO treatment (6.25%), folic acid treatment (70 µM), and a combination of VCO and folic acid. Variables assessed in this study included heart rate and insulin receptor substrate (Irs) were evaluated using real time polymerase chain reaction (RT-qPCR) analysis. Data were analyzed using SPSS version 27 for windows. Rotenone significantly reduced Irs expression compared with negative control (0.272 ± 0.128 vc 1.014 ± 0.187; p<0.001). Treatment with VCO (0.678 ± 0.250), folic acid (0.676 ± 0.191), and their combination (0.695 ±  0.231) increased Irs expression relative to the rotenone group (p<0.05), although no significant differences were observed among the treatment groups. Rotenone also elevated heart rate at 9 dpf (236.9 ± 19.2 bpm) compared with the negative control (163.7 ± 10.1 bpm), whereas treatment groups showed improved cardiac parameters. These findings highlight the potential of nutritional interventions targeting mitochondrial oxidative stress to support recovery of insulin signaling and improve metabolic stability in stunting related conditions.  

Keywords


Virgin Coconut Oil; Folic Acid; Insulin Signaling; Stunting; Zebrafish; Oxidative Stress; Heart rate

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Almansa-Ordonez, A., Bellido, R., Vassena, R., Barragan, M., & Zambelli, F. (2020). Oxidative stress in reproduction: A mitochondrial perspective. Biology, 9(9), 1–21. https://doi.org/10.3390/biology9090269

Arunima, S., & Rajamohan, T. (2014). Influence of virgin coconut oil-enriched diet on the transcriptional regulation of fatty acid synthesis and oxidation in rats – a comparative study. 1782–1790. https://doi.org/10.1017/S000711451400004X

Asbaghi, O., Ghanavati, M., Ashtary-larky, D., Bagheri, R., & Kelishadi, M. R. (2021). Effects of Folic Acid Supplementation on Oxidative Stress Markers : A Systematic Review and Meta-Analysis of Randomized Controlled Trials. 1–20.

Ayer, A., Fazakerley, D. J., James, D. E., & Stocker, R. (2022). The role of mitochondrial reactive oxygen species in insulin resistance. Free Radical Biology and Medicine, 179(November 2021), 339–362. https://doi.org/10.1016/j.freeradbiomed.2021.11.007

Chitnis, M. M., Yuen, J. S. P., Protheroe, A. S., Pollak, M., & Macaulay, V. M. (2008). The Type 1Insulin-Like Growth Factor Receptor Pathway. 14(20), 6364–6370. https://doi.org/10.1158/1078-0432.CCR-07-4879

Choi TY, Choi TI, Lee YR, Choe SK, K. C. (2021). Zebra fi sh as an animal model for biomedical research. 310–317. https://doi.org/10.1038/s12276-021-00571-5

Davila, M. P., Muñoz, P. M., Tapia, J. A., Ferrusola, C. O., Da Silva C, C. B., & Peña, F. J. (2015). Inhibition of mitochondrial complex I leads to decreased motility and membrane integrity related to increased hydrogen peroxide and reduced ATP production, while the inhibition of glycolysis has less impact on sperm motility. PLoS ONE, 10(9), 1–21. https://doi.org/10.1371/journal.pone.0138777

Dhalla, N. S., Elimban, V., Bartekova, M., & Adameova, A. (2022). Involvement of Oxidative Stress in the Development of Subcellular Defects and Heart Disease. Biomedicines, 10(2). https://doi.org/10.3390/biomedicines10020393

Díaz Del Moral, S., Benaouicha, M., Muñoz-Chápuli, R., & Carmona, R. (2021). The Insulin-like Growth Factor Signalling Pathway in Cardiac Development and Regeneration. International Journal of Molecular Sciences, 23(1). https://doi.org/10.3390/ijms23010234

Gonchar, O. O., Klymenko, O. O., Drevytska, T. I., Bratus, L. V, & Mankovska, I. M. (2021). oxidative stress in rat heart mitochondria under a rotenone model of parkinson’ disease: a corrective effect of capicor treatment. 93, 21–30.

Guo, C. A., & Guo, S. (2017). Insulin receptor substrate signaling controls cardiac energy metabolism and heart failure. Journal of Endocrinology, 233(3), R131–R143. https://doi.org/10.1530/JOE-16-0679

Heo, G., Sun, M., Jiang, W., Li, X., Lee, S., Guo, J., Zhou, D., & Id, X. C. (2022). Rotenone causes mitochondrial dysfunction and prevents maturation in porcine oocytes. 1–18. https://doi.org/10.1371/journal.pone.0277477

Hsu, H. C., Chiou, J. F., Wang, Y. H., Chen, C. H., Mau, S. Y., Ho, C. Te, Chang, P. J., Liu, T. Z., & Chen, C. H. (2013). Folate deficiency triggers an oxidative-nitrosative stress-mediated apoptotic cell death and impedes insulin biosynthesis in RINm5F pancreatic islet ?-cells: Relevant to the pathogenesis of diabetes. PLoS ONE, 8(11). https://doi.org/10.1371/journal.pone.0077931

Jeevanandam, J., San, Y., & Danquah, M. K. (2019). Zebrafish as a Model Organism to Study Nanomaterial Toxicity. 3(3), 195–208.

Khotimah, H., Arista, D. M., Amelia, R., Ratnaningrum, S. D., & Irwanto, Y. (2025). Rosmarinus officinalis and Centella asiatica in Gestational Diabetes Mellitus. 9(May), 2350–2356.

Kranenburg, E., Kubant, R., Cho, C. E., Yang, Z., Datars, M., Dong, J., & Anderson, G. H. (2025). Folic Acid Reduces Insulin Resistance in Mice With Diet-Induced Obesity by Altering One-Carbon Metabolism and DNA Methylation Patterns of Hypothalamic and Hepatic Insulin Receptor Gene. Molecular Nutrition and Food Research, 69(20), 1–11. https://doi.org/10.1002/mnfr.70181

Li, A., Zheng, N., & Ding, X. (2022). Mitochondrial abnormalities: a hub in metabolic syndrome-related cardiac dysfunction caused by oxidative stress. Heart Failure Reviews, 27(4), 1387–1394. https://doi.org/10.1007/s10741-021-10109-6

Liao, Y. H., Lin, J. G., Lin, C. C., Tsai, C. C., Lai, H. L., & Li, T. C. (2020). Traditional Chinese Medicine Treatment Associated with Female Infertility in Taiwan: A Population-Based Case-Control Study. Evidence-Based Complementary and Alternative Medicine, 2020. https://doi.org/10.1155/2020/3951741

Lin, J., Ma, H., Li, H., Han, J., Guo, T., Qin, Z., Jia, L., & Zhang, Y. (2022). The Treatment of Complementary and Alternative Medicine on Female Infertility Caused by Endometrial Factors. Evidence-Based Complementary and Alternative Medicine, 2022. https://doi.org/10.1155/2022/4624311

Luptak, I., Sverdlov, A. L., Panagia, M., Qin, F., Pimentel, D. R., Croteau, D., Siwik, D. A., Ingwall, J. S., Bachschmid, M. M., Balschi, J. A., & Colucci, W. S. (2018). Decreased ATP production and myocardial contractile reserve in metabolic heart disease. Journal of Molecular and Cellular Cardiology, 116(October 2017), 106–114. https://doi.org/10.1016/j.yjmcc.2018.01.017

Martínez Báez, A., Ayala, G., Pedroza-Saavedra, A., González-Sánchez, H. M., & Chihu Amparan, L. (2024). Phosphorylation Codes in IRS-1 and IRS-2 Are Associated with the Activation/Inhibition of Insulin Canonical Signaling Pathways. Current Issues in Molecular Biology, 46(1), 634–649. https://doi.org/10.3390/cimb46010041

Masenga, S. K., Kabwe, L. S., Chakulya, M., & Kirabo, A. (2023). Mechanisms of Oxidative Stress in Metabolic Syndrome. International Journal of Molecular Sciences, 24(9). https://doi.org/10.3390/ijms24097898

Mccluskey, B. M., & Braasch, I. (2020). Zebrafish Taxonomy and Phylogeny or Taxonomy and Phylogeny. In The Zebrafish in Biomedical Research: Biology, Husbandry, Diseases, and Research Applications. Elsevier. https://doi.org/10.1016/B978-0-12-812431-4.00002-6

Meijles, D. N., Cull, J. J., Markou, T., Cooper, S. T. E., Haines, Z. H. R., Fuller, S. J., Gara, P. O., Sheppard, M. N., Harding, S. E., Sugden, P. H., & Clerk, A. (2020). Activated Protein Kinase ) and Orchestrates Cardiac Remodeling to Hypertension. 1, 1208–1218. https://doi.org/10.1161/HYPERTENSIONAHA.119.14556

Menezo, Y., Elder, K., Clement, A., & Clement, P. (2022). Folic Acid, Folinic Acid, 5 Methyl TetraHydroFolate Supplementation for Mutations That Affect Epigenesis through the Folate and One-Carbon Cycles. Biomolecules, 12(2). https://doi.org/10.3390/biom12020197

Moh, J. (2022). Incorporation of Oxidized Phenylalanine Derivatives into Insulin Signaling Relevant Proteins May Link Oxidative Stress to Signaling Conditions Underlying Chronic Insulin Resistance.

Nurdiana, N., Claudia, C., Sartika, D., Anggriani, D. D., Luh, N., Mastuti, P., Ariani, A., & Kusuma, I. D. (2025). Tropical Journal of Natural Product Research Impact of Virgin Coconut Oil ( VCO ) on Locomotor Speed and Bax Expression in Zebrafish ( Danio rerio ) Larvae Stunting Model. 9(November), 5306–5311.

Nurdiana, N., Sartika, D., Anggriani, D. D., Claudia, C., Mastuti, N. L. P. H., Ariani, A., & Kusuma, I. D. (2025). Tropical Journal of Natural Product Research The Effect of Virgin Coconut Oil on IL-6 Gene Expression and Body Length in Stunted Zebrafish ( Danio rerio ) Larvae. 9(August), 3839–3844.

Prendergast, A. J., Humphrey, J. H., Prendergast, A. J., & Humphrey, J. H. (2014). Paediatrics and International Child Health The stunting syndrome in developing countries The stunting syndrome in developing countries. 9047. https://doi.org/10.1179/2046905514Y.0000000158

Primihastuti, D., Woro, D., Kusumo, K., Primaditya, V., Arsy, F., Cory, N., Ika, L., Ariati, P., Kalsum, U., Khotimah, H., Ali, M., & Riawan, W. (2022). The effect of ethanol extract of pegagan ( Centella asiatica ) on bone ossification and osteoclastogenesis on the stunting model of Zebrafish ( Danio rerio ) larvae induced by rotenone.

Pusdatin. (2022). Infodatin Keluarga Bebas Stunting.

Rains, J. L., & Jain, S. K. (2011). Free Radical Biology & Medicine Oxidative stress , insulin signaling , and diabetes. Free Radical Biology and Medicine, 50(5), 567–575. https://doi.org/10.1016/j.freeradbiomed.2010.12.006

Schraps, N., Tirre, M., Pyschny, S., Reis, A., Schlierbach, H., Seidl, M., Kehl, H. G., Schänzer, A., Heger, J., Jux, C., & Drenckhahn, J. D. (2024). Cardiomyocyte maturation alters molecular stress response capacities and determines cell survival upon mitochondrial dysfunction. Free Radical Biology and Medicine, 213(November 2023), 248–265. https://doi.org/10.1016/j.freeradbiomed.2024.01.034

Shah, A. K., Bhullar, S. K., Elimban, V., & Dhalla, N. S. (2021). Oxidative stress as a mechanism for functional alterations in cardiac hypertrophy and heart failure. Antioxidants, 10(6). https://doi.org/10.3390/antiox10060931

Sherer, T. B., Betarbet, R., Testa, C. M., Seo, B. B., Richardson, J. R., Kim, J. H., Miller, G. W., Yagi, T., Matsuno-yagi, A., & Greenamyre, J. T. (2003). Mechanism of Toxicity in Rotenone Models of Parkinson ’ s Disease. 23(34), 10756–10764.

Son, Y., Cheong, Y., Kim, N., Chung, H., Kang, D. G., & Pae, H. (2011). Mitogen-Activated Protein Kinases and Reactive Oxygen Species : How Can ROS Activate MAPK Pathways ? 2011. https://doi.org/10.1155/2011/792639

Sumiyati, Y. (2024). Colocasia esculentaL. EXTRACT INCREASE GROWTH AND MOTILITYOF ROTENONE INDUCED ZEBRAFISH (Danio rerio) LARVAE STUNTING MODEL. 12(1), 633–642.

Thimmana Gouda B1, M Kumar , S Geethanjali3, V. V. & J. S., & 1Department. (2024). e - Publishing Group A comprehensive review of virgin coconut oil : Extraction methods and diverse applications. 11(2), 1–10.

Toyoshima, Y., Nakamura, K., Taguchi, Y., Tokita, R., Takeuchi, S., Osawa, H., Teramoto, N., Sugihara, H., Yoshizawa, F., Yamanouchi, K., & Minami, S. (2025). Deletion of IRS-1 leads to growth failure and insulin resistance with downregulation of liver and muscle insulin signaling in rats. 1–15.

Wang, L., Duan, Q., Wang, T., Ahmed, M., Zhang, N., Li, Y., Li, L., & Yao, X. (2015). Mitochondrial Respiratory Chain Inhibitors Involved in ROS Production Induced by Acute High Concentrations of Iodide and the Effects of SOD as a Protective Factor. 2015. https://doi.org/10.1155/2015/217670

Wati, S. C. (2018). PENGARUH PEMBERIAN ASAM FOLAT FASE PRENATAL TERHADAP PANJANG BADAN DAN FREKUENSI DETAK JANTUNG PADA LARVA ZEBRAFISH MODEL STUNTING DENGAN INDUKSI ROTENON TUGAS AKHIR Untuk Memenuhi Persyaratan Memperoleh Gelar Sarjana Kedokteran OLEH : Chandra Dewi Sarasw.

WHO. (2023). Stunting prevalence among children under 5 years of age (% height-for-age < -2 SD ), survey-based estimates Year. https://www.who.int/data/gho/data/indicators/indicator-details/GHO/gho-jme-country-children-aged-5-years-stunted-%28-height-for-age--2-sd%29

Wood A.W., D. C. dan B. H. A. (2007). Insulin-like growth factor-binding protein-1 : an evolutionarily conserved fine tuner of insulin-like growth factor action under catabolic and stressful conditions. 71, 309–325. https://doi.org/10.1111/j.1095-8649.2007.01606.x

Woro, D., Kusumo, K., & Primihastuti, D. (2020). Ethanolic Extract of Centella asiatica Increased IGF-1 and IRS Expression in Zebrafish (Danio rerio) Rotenone-induced. June 2019.

Zhao, Y., Zhang, Y., Liu, D., Feng, H., Wang, X., Su, J., Yao, Y., Ng, E. H. Y., Yeung, W. S. B., Li, R. H. W., Rodriguez-Wallberg, K. A., & Liu, K. (2022). Identification of curcumin as a novel potential drug for promoting the development of small ovarian follicles for infertility treatment. PNAS Nexus, 1(3), 1–12. https://doi.org/10.1093/pnasnexus/pgac108




DOI: https://doi.org/10.14421/biomedich.2026.151.199-207

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Copyright (c) 2026 Syahana Aini, Farica Emiliana, Hanida Aisyah Ardiana, Nurdiana, Ariani, Brigitta Ida Resita Vebrianti Corebima



Biology, Medicine, & Natural Product Chemistry
ISSN 2089-6514 (paper) - ISSN 2540-9328 (online)
Published by Sunan Kalijaga State Islamic University & Society for Indonesian Biodiversity.

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