The Effect of Virgin Coconut Oil on IL-6 Gene Expression and Body Length in Stunted Zebrafish (Danio rerio) Larvae

Nurdiana Nurdiana; Dian Sartika; Dina Anggriani; Claudia Claudia; Ni Mastuti; Ariani Ariani; Takhta Khalasha; Ihda Kusuma

doi:10.26538/tjnpr/v9i8.44

Authors

Nurdiana Nurdiana Departement of Pharmacology, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Dian Sartika Master Program of Midwifery, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Dina D Anggriani Master Program of Midwifery, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Claudia Claudia Master Program of Midwifery, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Ni Luh P H Mastuti Departement of Midwifery, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Ariani Ariani Departement of Pediatrics, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Takhta Khalasha Departement of Pharmacology, Faculty of Medicine, Brawijaya University, Malang, Indonesia
Ihda D Kusuma Departement of Pathological Anatomy, Faculty of Medicine, Brawijaya University, Malang, Indonesia

DOI:

https://doi.org/10.26538/tjnpr/v9i8.44

Keywords:

growth, rotenone, body length, Interleukin-6, zebrafish model, stunting, Virgin Coconut Oil

Abstract

Stunting poses a major global health concern, impairing physical and cognitive development, particularly in developing nations; it persists despite various nutrition initiatives. Virgin Coconut Oil (VCO) derived from local resources is emerging as an effective option to enhance nutrition and promote brain and motor skills growth in children suffering from stunting. The objective of this study is to evaluate the effect of VCO on the expression of Interleukin-6 (IL-6) and body length in zebrafish larvae induced with stunting by rotenone exposure. Zebrafish larvae 2 hours post-fertilization were treated with rotenone to induce stunting and subsequently administered VCO at various concentrations. IL-6 expression was measured using the Reverse Transcription Polymerase Chain Reaction (RT-PCR) method, and body length was measured at 3, 6, and 9 days post-fertilization. The results of the study showed that the body length of zebrafish larvae exposed to rotenone significantly decreased compared to the negative control group, measuring 3.04 mm on day 3 and 3.72 mm on day 9. Administration of VCO at a dose of 6.25% was able to increase the body length of the larvae to 3.32 mm on day 3 and 4.06 mm on day 9. The expression of the IL-6 gene, statistical analysis using one-way ANOVA revealed a highly significant difference among the groups (p < 0.001). These findings indicate that VCO effectively reduces inflammation and improves growth inhibited by rotenone exposure.

References

⦁ Siagian FE. Role of Intestinal Parasitic Infection in Stunting. South Asian J Res Microbiol. 2023;15(2):41–50.

⦁ Pelosi L, Berardinelli MG, Forcina L, Ascenzi F, Rizzuto E, Sandri M, Benedetti FD, Scicchitano BM and Musaro A. Sustained systemic levels of IL-6 impinge early muscle growth and induce muscle atrophy and wasting in adulthood. Cells. 2021;10(7):1–15.

⦁ Cansiz D, Unal I, Ustundag UV, Alturfan AA, A.Altinoz M, Elmacr I, Alturfan EE . Caprylic acid ameliorates rotenone induced inflammation and oxidative stress in the gut-brain axis in Zebrafish. Mol Biol Rep. 2021;48(6):5259–5273.

⦁ Iswati RS, Indria Nuraini. Effectiveness of Virgin Coconut Oil (VCO) on Changes in Weight and Height among Under-five Children with Stunting. Embrio J Kebidanan. 2023;15(1):112–118.

⦁ Khotimah H, Alfred T, Wangta C, Chandra K, Tjan B, Christianto A, Ali M, Kalsum U. Miracle tree (Moringa oleifera) extract prevents rotenone-induced stunting in zebrafish (Danio rerio) through decreasing caspase-3. Int Conf Life Sci Technol. 2021;030003.

⦁ Sorribes A, Porsteinsson H, Arnardóttir H, Jóhannesdóttir IP, Sigurgeirsson B, de Polavieja GG, Karlsson KAE. The ontogeny of sleep-wake cycles in zebrafish: A comparison to humans. Front Neural Circuits. 2013;7(NOV):1–13.

⦁ Cory’ah FAN, Primaditya V, Ariati LIP, Zakiah, Wardani DWKK, Primihastuti D, Yuningsih, Khotimah H, Ali MM, Nurdiana. Ethanolic extract of centella asiatica increased IGF-1 and IRS expression in zebrafish (Danio rerio) rotenone-induced. AIP Conf Proc. 2020;2231(April).

⦁ Christianto A, Khotimah H, Kalsum U. The Effect of Moringa oleifera Leaf Extract on Body Length, Malondialdehyde Levels, and Interleukin-6 in Zebrafish (Danio rerio) Larvae as a Stunting Model Induced by Rotenone.2018;1–114. Available from: https://repository.ub.ac.id/id/eprint/195285/

⦁ Vanhauwaert S, Lefever S, Coucke P, Speleman F, De Paepe A, Vandesompele J, Willaert A. RT-qPCR gene expression analysis in zebrafish: Preanalytical precautions and use of expressed repetitive elements for normalization. Methods Cell Biol. 2016;135:329–342.

⦁ Yuningsih,Wardani DWKK, Ali M, Khotimah H, Nurdiana, Riawan W, Primihastuti D, Cory'ah FAN, Ariati LIP, Primaditya V, Zakiyah. The effect of Centella asiatica ethanolic extract on expression of glucose transporter 1 and osteocalcin on stunting larvae of Zebrafish (Danio rerio). GSC Biol Pharm Sci. 2022;19(3):178–189.

⦁ Wijayanti AR, Ridlayanti A, Ali HKM. Protection of Pegagan (Centella asiatica) Extract Through Hsp60 and Bax Expression on Stunting Model Zebrafish Larvae (Danio rerio) By Rotenone Inducted.pdf. IOP Science; 2021.

⦁ Primaditya V, Cory’ah FAN, Ariati LIP, Zakiah, Wardani DWKK, Yuningsih, Primihastuti D, Khotimah H, Ali MM, Riawan W. Effect of centella asiatica to the glucose transporter 4 and osteocalcin on the rotenonee-induced zebrafish larvae (Danio rerio) stunting model. AIP Conf Proc. 2020;2231.

⦁ Selvaraj KV, Bharathi A, Sivakumar V, Karthikeyan A, Maheswarappa H. Virgin coconut oil: A nutraceutical and therapeutic food. Int J Chem Stud. 2020;8(3):662–663.

⦁ Kusuma MA, Putri NA. Review: Fatty Acids of Virgin Coconut Oil (VCO) and Their Health Benefits. J Agrinika: Journal of Agrotechnology and Agribusiness.2020;4(1):93.

⦁ Rohman A, Irnawati, Erwanto Y, Lukitaningsih E, Rafi M, Fadzilah NA, Windarsih A, Sulaiman A, Zakaria Z. Virgin Coconut Oil: Extraction, Physicochemical Properties, Biological Activities and Its Authentication Analysis. Food Rev Int [Internet]. 2021;37(1):46–66. Available from: https://doi.org/10.1080/87559129.2019.1687515

⦁ Álvarez-Martínez FJ, Barrajón-Catalán E, Herranz-López M, Micol V. Antibacterial plant compounds, extracts and essential oils: An updated review on their effects and putative mechanisms of action. Phytomedicine. 2021;90.

⦁ Roopashree PG, Shetty SS, Suchetha Kumari N. Effect of medium chain fatty acid in human health and disease. J Funct Foods [Internet]. 2021;87:104724. Available from: https://doi.org/10.1016/j.jff.2021.104724

⦁ Kamri AM. Review: Virgin Coconut Oil and The Pharmacology Effect. J Pharmascience. 2024;11(1):115.

⦁ Asish A, Rajan S, Jose SP, Sheethal S, Ratheesh M, Sandya S. Extracellular Modulatory and Anti-Inflammatory Effects of a Combiation of Curcumin, Piperine, and Virgin Coconut Oil in Animal Model. Asian J Pharm Clin Res. 2024;17(12):73–79.

⦁ Chen A Di, Cao JX, Chen HC, Du HL, Xi XX, Sun J, Yin J, Jing YH, Gao LP. Rotenone aggravates PD-like pathology in A53T mutant human α-synuclein transgenic mice in an age-

dependent manner. Front Aging Neurosci. 2022;14:1–15.

⦁ Azimullah S, Jayaraj RL, Meeran MFN, Jalal FY, Adem A, Ojha S, Beiram R. Myrcene Salvages Rotenone-Induced Loss of Dopaminergic Neurons by Inhibiting Oxidative Stress, Inflammation, Apoptosis, and Autophagy. Molecules. 2023;28(2):1–14.

⦁ Widjaja NA, Hamidah A, Purnomo MT, Ardianah E. Effect of lactoferrin in oral nutrition supplement (ONS) towards IL-6 and IL-10 in failure to thrive children with infection. F1000Research. 2023;12:1–40.

⦁ Eddin LB, Azimullah S, Jha NK, Nagoor Meeran MF, Beiram R, Ojha S. Limonene, a Monoterpene, Mitigates Rotenone-Induced Dopaminergic Neurodegeneration by Modulating Neuroinflammation, Hippo Signaling and Apoptosis in Rats. Int J Mol Sci. 2023;24(6).

⦁ Zambruni M, Ochoa TJ, Somasunderam A, Cabada MM, Morales ML, Mitreva M, Rosa BA, Acosta AJ, Riveros M, Arango S, Durand D, Berends MN, Melby P, Utay NS. Stunting is preceded by intestinal mucosal damage and microbiome changes and is associated with systemic inflammation in a cohort of Peruvian infants. Am J Trop Med Hyg. 2019;101(5):1009–1017.

⦁ Anusha G, Sunayana R, Ponnam M, Kumar BA. Asian Journal of Pharmaceutical Research and Development. Asian J Pharm Res Dev. 2020;8(6):77–80.

⦁ Jack KS, Asaruddin MR Bin, Bhawani SA. Pharmacophore study, molecular docking and molecular dynamic simulation of virgin coconut oil derivatives as anti-inflammatory agent against COX-2. Chem Biol Technol Agric [Internet]. 2022;9(1):1–26. Available from: https://doi.org/10.1186/s40538-022-00340-0

⦁ Zeng YQ, He JT, Hu BY, Li W, Deng J, Lin QL. Virgin coconut oil: A comprehensive review of antioxidant activity and mechanisms contributed by phenolic compounds. Crit Rev Food Sci Nutr. 2024;64(4):1052–1075.

⦁ Olędzka AJ, Czerwińska ME. Role of Plant-Derived Compounds in the Molecular Pathways Related to Inflammation. Int J Mol Sci. 2023;24(5).

⦁ Ariani A, Ghofar IF, Khotimah H, Nurdiana N, Rahayu M. Asiatic acid in Centella asiatica extract towards morphological development in an intermittent hypoxia intrauterine embryo model and molecular prediction pathway of insulin-like growth factor-1 receptor signalling. Open Vet J. 2023;13(5):629–637.

⦁ Okolie NP, Falodun A, Davids O. Evaluation of the Antioxidant Activity of Root Extract of Pepper Fruit (Dennetia Tripetala), and It’S Potential for the Inhibition of Lipid Peroxidation. African J Tradit Complement Altern Med. 2014;11(3):221–227.

⦁ Ariani A, Khotimah H, Sulistyarini A, Daniaridevi AS. Centella asiatica extract ameliorates deoxygenation-induced neurological dysfunction in zebrafish larvae. Open Vet J. 2024;14(5):1154–1160.

⦁ Nikmah UK, Pertiwi D, Hussaana A. The Effect of Virgin Coconut Oil (VCO) on Malondialdehyde, Glutathione Peroxidase, and Cortisol Hormone Levels. Int J Multidiscip Res Anal. 2023;06(03):897–903.

⦁ Kandiyil SP, Jose A, Mohanan C, Illam SP, Raghavamenon AC. Virgin coconut oil mitigates ageing-associated oxidative stress and dyslipidaemia in male Wistar rats. Nutr Metab Cardiovasc Dis [Internet]. 2024;34(12):2834–2841. Available from: https://doi.org/10.1016/j.numecd.2024.08.012

⦁ Fadlalla EA, Seddik A. Anti-Inflammatory and Nephroprotective Influence of Virgin Coconut Oil on Gentamicin-Induced Nephrotoxicity in Nondiabetic and Streptozotocin-Induced Diabetic Rats. African J Food, Agric Nutr Dev. 2023;23(2):22492–22512.