Rasbora borneensis fish and Vigna unguiculata legume protein supplementation restores reproductive hormone profiles in early-life protein-deficient female rats

  • Didik Dwi Sanyoto Anatomy Laboratory, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarmasin https://orcid.org/0000-0001-8006-6787
  • Triawanti Biochemistry and Biomolecular Laboratory, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarmasin https://orcid.org/0000-0002-5753-3620
  • Sherly Limantara Department of Psychiatry, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarmasin.
  • Juhairana Department of Nutritional Sciences, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarmasin. https://orcid.org/0009-0008-2218-6439
  • Nani Zaitun Department of Internal Medicine, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarmasin.
  • Haitami Doctoral Program, Medical Science Study Program, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarbaru
  • Dimas Ikhsan Airlangga Biochemistry and Biomolecular Laboratory, Faculty of Medicine and Health Sciences, Lambung Mangkurat University, Banjarmasin. https://orcid.org/0000-0002-5044-4385
  • Metria Zeta Students of the Medical Study Program, Undergraduate Program, Faculty of Medicine, Lambung Mangkurat University, Banjarmasin.
  • Muhammad Athaya Zein Students of the Medical Study Program, Undergraduate Program, Faculty of Medicine, Lambung Mangkurat University, Banjarmasin.

Abstract

Background: Protein malnutrition during early development can alter the hormonal pathways essential for female reproductive maturation. It remains a significant concern in regions where inadequate protein intake contributes to stunting and long-term reproductive risks. However, evidence remains limited regarding how maternal and early-life protein deficiency affects reproductive hormone formation in offspring, and whether locally available nutrient sources can reverse these disruptions.

Objective: This study evaluated the effects of supplementation of Rasbora borneensis and Vigna unguiculata spp. cylindrica on reproductive hormones in female rat offspring subjected to protein deficiency.

Methods: A protein-deficient model was generated by feeding rats a low-protein AIN-76A diet from birth to weaning, followed by continuation of the same diet in female offspring for four weeks. The offspring were then assigned to a normal control group receiving a standard diet, a negative control group maintained on the low-protein diet, and three treatment groups receiving Seluang fish feed, cowpea cowpea feed, or a combination of both for four weeks. FSH and estrogen levels were assessed to evaluate reproductive endocrine function.

Results: All treatment diets increased FSH and estrogen levels compared with the negative control. FSH concentrations were significantly higher in all treatment groups (p = 0.012), with the combination diet yielding the greatest increase. Estrogen levels also rose substantially in the treatment groups with statistical analyses confirming significant differences (p < 0.001).

Conclusion: Seluang fish and Nagara cowpea effectively improved reproductive hormone profiles in protein-deficient female rats, highlighting their potential as relevant nutritional strategies to counter early-life protein malnutrition.

References

Calcaterra V, Verduci E, Stagi S, Zuccotti G. How the intricate relationship between nutrition and hormonal equilibrium significantly influences endocrine and reproductive health in adolescent girls. Front Nutr. 2024;11: 1337328. https://doi.org/10.3389/fnut.2024.1337328

Kemenkes. Buku Saku Hasil Survei Status Gizi Indonesia (SSGI) 2022. BKPK; 2022.

Sari AN, Diana FM, Nur NC. The association between protein intake and feeding practice with stunting in children aged 24-59 months. Nutr clin diet hosp. 2025;45. https://doi.org/10.12873/451diana

Haryani VM, Putriana D, Hidayati RW. Animal-Based Protein Intake is Associated with Stunting in Children in Primary Health Care of Minggir. Amerta Nutr. 2023;7: 139-146. https://doi.org/10.20473/amnt.v7i2SP.2023.139-146

Tang X, Zhao Y, Liu Q, Hu D, Li G, Sun J, et al. The effect of risk accumulation on childhood stunting: A matched case-control study in China. Front Pediatr. 2022;10: 816870. https://doi.org/10.3389/fped.2022.816870

De Sanctis V, Soliman A, Alaaraj N, Ahmed S, Alyafei F, Hamed N. Early and long-term consequences of nutritional stunting: From childhood to adulthood. Acta Biomed. 2021;92: e2021168.

Vargas R, Martins IP, Matiusso CCI, Casagrande RA, Zara CB, de Souza AC, et al. Protein restriction during lactation causes transgenerational metabolic dysfunction in adult rat offspring. Front Nutr. 2022;9: 1062116. https://doi.org/10.3389/fnut.2022.1062116

Benabbou M, Allaoui A, Zahzeh AKM, Boualga A, Zahzeh T. Protein vs. Energy restriction impact on rat testis' function, redox status, and histomorphometry. Prev Nutr Food Sci. 2023;28: 61-68. https://doi.org/10.3746/pnf.2023.28.1.61

Cao Y, Yang M, Song J, Jiang X, Xu S, Che L, et al. Dietary protein regulates female estrous cyclicity partially via fibroblast growth factor 21. Nutrients. 2023;15: 3049. https://doi.org/10.3390/nu15133049

Khorram O, Keen-Rinehart E, Chuang T-D, Ross MG, Desai M. Maternal undernutrition induces premature reproductive senescence in adult female rat offspring. Fertil Steril. 2015;103: 291-8.e2. https://doi.org/10.1016/j.fertnstert.2014.09.026

Spicer J, Malaspina D, Blank S V, Goosens KA. Follicle-stimulating hormone: More than a marker for menopause: FSH as a frontier for women's mental health. Psychiatry Res. 2025;345: 116239. https://doi.org/10.1016/j.psychres.2024.116239

Liu W, Xin Q, Wang X, Wang S, Wang H, Zhang W, et al. Estrogen receptors in granulosa cells govern meiotic resumption of pre-ovulatory oocytes in mammals. Cell Death Dis. 2017;8: e2662-e2662. https://doi.org/10.1038/cddis.2017.82

Valera H, Chen A, Grive KJ. The hypothalamic-pituitary-ovarian axis, ovarian disorders, and brain aging. Endocrinology. 2025;166. https://doi.org/10.1210/endocr/bqaf137

Lopes CVA, de Sousa Alves Neri JL, Hunter J, Ronto R, Mihrshahi S. Interventions and Programs Using Native Foods to Promote Health: A Scoping Review. Nutrients. 2024;16: 4222. https://doi.org/10.3390/nu16234222

Triawanti T, Yunanto A, Dwi Sanyoto D, Wana Nuramin H. Nutritional Status Improvement in Malnourished Rat (Rattus norvegicus) after Seluang Fish (Rasbora spp.) Treatment. Curr Res Nutr Food Sci J. 2018;6: 127-134. https://doi.org/10.12944/CRNFSJ.6.1.14

Hustiany R, Rahmawati E, Rahmi A. Development potential of nagara bean (vigna unguiculata ssp. Cylindrica) cultivated in freshwater swamplands for processed food. Tropical Wetland Journal. 2016;2: 30-36. https://doi.org/10.20527/twj.v2i3.37

Wulansari A, Hidayat AT, Wisudariani E, Asty ZF. The potential of fish powder Seluang fish (Rasbora argytaenia) as a source of protein in complementary feeding for toddlers. Proceedings Academic Universitas Jambi. 2025. pp. 848-853.

Mohanty B, Mahanty A, Ganguly S, Sankar T V, Chakraborty K, Rangasamy A, et al. Amino Acid compositions of 27 food fishes and their importance in clinical nutrition. J Amino Acids. 2014;2014: 269797. https://doi.org/10.1155/2014/269797

Salamanca N, Herrera M, de la Roca E. Amino Acids as Dietary Additives for Enhancing Fish Welfare in Aquaculture. Animals. 2025;15: 1293. https://doi.org/10.3390/ani15091293

Aragão C, Engrola S, Costas B. Amino acid supplementation in fish nutrition and welfare. Animals (Basel). 2025;15: 1656. https://doi.org/10.3390/ani15111656

Hustiany R. Sustainability of protein potential in nagara beans (Vigna unguiculata ssp Cylindrica) from South Kalimantan. IOP Conf Ser Earth Environ Sci. 2024;1302: 12085. https://doi.org/10.1088/1755-1315/1302/1/012085

Shea Z, Ogando do Granja M, Fletcher EB, Zheng Y, Bewick P, Wang Z, et al. A Review of Bioactive Compound Effects from Primary Legume Protein Sources in Human and Animal Health. Curr Issues Mol Biol. 2024;46: 4203-4233. https://doi.org/10.3390/cimb46050257

da Silva A, de Freitas Barbosa M, da Silva P, de Oliveira J, da Silva T, Lopes Teixeira Junior D, et al. Health benefits and industrial applications of functional Cowpea seed proteins. Grain and Seed Proteins Functionality. IntechOpen; 2021.

Setyarini AI, Barlianto W, Nugrahenny D, Winarsih S. Analysis of Phytoestrogen Compounds from Vigna Unguiculata Using LC-MS/MS Approach. Advanced Journal of Chemistry. 2025;8: 1431-1441.

Fithri AN, Yueniwati Y, Arsana IW, Khotimah H, Nurwidyaningtyas W. The potential health benefit of Cowpea estrogen-like activity to restore vaginal epithelial cells thinning due to menopausal syndrome. Trop J Nat Prod Res. 2024;8: 6624-6630. https://doi.org/10.26538/tjnpr/v8i3.22

Sombié PAED, Compaoré M, Coulibaly AY, Ouédraogo JT, Tignégré J-BDLS, Kiendrébéogo M. Antioxidant and phytochemical studies of 31 cowpeas (Vigna unguiculata (Walp L)) genotypes from Burkina. Foods. 2018;7: 143. https://doi.org/10.3390/foods7090143

Padhi SR, Bartwal A, John R, Tripathi K, Gupta K, Wankhede DP, et al. Evaluation and multivariate analysis of Cowpea [Vigna unguiculata (L.) Walp] germplasm for selected nutrients-mining for nutri-dense accessions. Front Sustain Food Syst. 2022;6. https://doi.org/10.3389/fsufs.2022.888041

Jayathilake C, Visvanathan R, Deen Afka and Bangamuwage R, Jayawardana BC, Nammi S, Liyanage R. Cowpea: an overview on its nutritional facts and health benefits. J Sci Food Agric. 2018;98: 4793-4806. https://doi.org/10.1002/jsfa.9074

Shevkani K, Shivani B, Dhaka SS, Patil C. Cowpeas for sustainable agriculture and nutrition security: an overview of their nutritional quality and agroeconomic advantages. Discov Food. 2025;5. https://doi.org/10.1007/s44187-025-00382-x

Augustin V, Badanthadka M, R J M, Dsouza V, Kumar BM, Shetty AV. Longitudinal evaluation of developmental protein malnutrition resembling marasmic-kwashiorkor condition in Wistar rats. Turk J Pharm Sci. 2024;21: 474-482.

Abey NO, Ebuehi OAT, Imaga NOA. Intergenerational protein deficiency and adolescent reproductive function of subsequent female generations (F1 and F2) in rat model. Curr Res Physiol. 2022;5: 16-24. https://doi.org/10.1016/j.crphys.2021.12.003

Picó C, Palou M, Pomar CA, Rodr'iguez AM, Palou A. Leptin as a key regulator of the adipose organ. Rev Endocr Metab Disord. 2022;23: 13-30. https://doi.org/10.1007/s11154-021-09687-5

Guzmán C, Cabrera R, Cárdenas M, Larrea F and Nathanielsz PW, Zambrano E. Protein restriction during fetal and neonatal development in the rat alters reproductive function and accelerates reproductive ageing in female progeny. J Physiol. 2006;572: 97-108. https://doi.org/10.1113/jphysiol.2005.103903

Sathishkumar K, Elkins R, Yallampalli U, Yallampalli C. Protein restriction during pregnancy induces hypertension in adult female rat offspring-influence of oestradiol. Br J Nutr. 2012;107: 665-673. https://doi.org/10.1017/S0007114511003448

Blakemore J, Naftolin F. Aromatase: Contributions to physiology and disease in women and men. Physiology (Bethesda). 2016;31: 258-269. https://doi.org/10.1152/physiol.00054.2015

Metallinou C, Staneloudi C, Nikolettos K, Asimakopoulos B. NGF, EPO, and IGF-1 in the male reproductive system. J Clin Med. 2024;13: 2918. https://doi.org/10.3390/jcm13102918

Deer E, LaMarca B, Reckelhoff Jane F and Shawky NM, Edwards K. The role of mitochondrial dysfunction and oxidative stress in women's reproductive disorders: Implications for polycystic ovary syndrome and preeclampsia. Int J Mol Sci. 2025;26: 6439. https://doi.org/10.3390/ijms26136439

Ju W, Yan B, Li D, Lian F, Xiang S. Mitochondria-driven inflammation: a new frontier in ovarian ageing. J Transl Med. 2025;23: 1005. https://doi.org/10.1186/s12967-025-06966-6

Gorzkiewicz J, Bartosz G, Sadowska-Bartosz I. The potential effects of phytoestrogens: The role in neuroprotection. Molecules. 2021;26: 2954. https://doi.org/10.3390/molecules26102954

van Duursen MBM. Modulation of estrogen synthesis and metabolism by phytoestrogensin vitroand the implications for women's health. Toxicol Res (Camb). 2017;6: 772-794. https://doi.org/10.1039/c7tx00184c

Lephart ED. Modulation of aromatase by phytoestrogens. Enzyme Res. 2015;2015: 594656. https://doi.org/10.1155/2015/594656

Dom'inguez-López I, Yago-Aragón Maria and Salas-Huetos A, Tresserra-Rimbau A, Hurtado-Barroso S. Effects of dietary phytoestrogens on hormones throughout a human lifespan: A review. Nutrients. 2020;12: 2456. https://doi.org/10.3390/nu12082456

Olaniyan EJ, Emokpae MA, Oyakhire FO, Adeagbo AL, Esezobor IK, Olaniyan SO. Impact of soybean phytoestrogen-rich extract on markers of inflammation markers in 4-vinyl cyclohexane diepoxide-induced menopause in albino rats. Med Lab Technol J. 2023. https://doi.org/10.31964/mltj.v9i2.549

Garner TB, Hester JM, Carothers A, Diaz FJ. Role of zinc in female reproduction. Biol Reprod. 2021;104: 976-994. https://doi.org/10.1093/biolre/ioab023

Zia-Ul-Haq M, Ahmad S, Amarowicz Ryszard and De Feo V. Antioxidant activity of the extracts of some cowpea (Vigna unguiculata (L) Walp.) cultivars commonly consumed in Pakistan. Molecules. 2013;18: 2005-2017. https://doi.org/10.3390/molecules18022005

Published
2026-02-06
How to Cite
Sanyoto, D. D., Triawanti, Limantara, S., Juhairana, Zaitun, N., Haitami, Airlangga, D. I., Zeta, M., & Zein, M. A. (2026). Rasbora borneensis fish and Vigna unguiculata legume protein supplementation restores reproductive hormone profiles in early-life protein-deficient female rats. Acta Biochimica Indonesiana, 9(1), 227. https://doi.org/10.32889/actabioina.227