Subchronic toxicological evaluation of the Holothuria atra extract on the histopathology of digestive organs of rats (Rattus norvegicus)

Muhammad Nursid Endar Marraskuranto Ariyanti S. Dewi Gintung Patantis Tiara Silva Khatulistiani Yann Yeo Sri Estuningsih   

Open Access   

Published:  Mar 10, 2025

DOI: 10.7324/JAPS.2025.216477
Abstract

The objective of this study was to determine the effect of ethanolic extract of sea cucumber Holothuria atra on the digestive organs of Rattus norvegicus rats for 90 days. The experimental rats were divided into six groups, namely, group A (control), group B (dose of 25-mg/kg BW), group C (50-mg/kg BW), group D (100-mg/kg BW), group E (200-mg/kg BW), and group F (400-mg/kg BW). Histopathological studies of the small intestine, pancreas, and stomach were conducted after 90 days. According to the investigation, the extract from H. atra was not harmful to the small intestine, pancreas, or stomach. The feed of H. atra extract at a dose of 50-mg/kg BW increased gastric gland cells, reduced damage to pancreatic acinar cells, increased the height and surface area of villi, and increased the number of small intestinal crypts. Ethanolic extract at this dose also showed an effect on reducing the number of inflammatory cells in the pancreas and small intestinal submucosa and maintaining the surface of the gastric mucosa by reducing lesions. The H. atra ethanolic extract was considered safe for the digestive organs of R. norvegicus rats. Apart from being safe, at the right dose, H. atra extract provides benefits for digestive organs.


Keyword:     Subchronic toxicity digestive organs Rattus norvegicus sea cucumber Holothuria atra


Citation:

Nursid M, Marraskuranto E, Dewi AS, Patantis G, Khatulistiani TS, Yeo Y, Estuningsih S. Subchronic toxicological evaluation of the Holothuria atra extract on the histopathology of digestive organs of rats (Rattus norvegicus). J Appl Pharm Sci. 2025. Online First. http://doi.org/10.7324/JAPS.2025.216477

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

HTML Full Text

Reference

1. Sensoy I. A review on the food digestion in the digestive tract and the used in vitro models. Curr Res Food Sci. 2021;4:308–19. Available from: https://www.sciencedirect.com/science/article/pii/S2665927121000307

2. Zimmermann C, Wagner A. Impact of food-derived bioactive compounds on intestinal immunity. Biomolecules. 2021;11(12):1901. Available from: https://www.mdpi.com/2218-273X/11/12/1901

3. Xu T, Sun R, Zhang Y, Zhang C, Wang Y, Wang Z, et al. Recent research and application prospect of functional oligosaccharides on intestinal disease treatment. Molecules [Internet]. 2022;27(21):7622. Available from: https://www.mdpi.com/1420-3049/27/21/7622

4. Essa M, Bishir M, Bhat A, Chidambaram S, Al-Balushi B, Hamdan H, et al. Functional foods and their impact on health. J Food Sci Technol [Internet]. 2023;60(3):820–34. doi: https://doi.org/10.1007/s13197-021-05193-3

5. Yan T, Babji A, Lim S, Sarbini S. A systematic review of edible swiftlet’s nest (ESN): nutritional bioactive compounds, health benefits as functional food, and recent development as bioactive ESN glycopeptide hydrolysate. Trends Food Sci Technol [Internet]. 2021;115:117–32. Available from: https://www.sciencedirect.com/science/article/pii/S0924224421004088

6. Hossain A, Dave D, Shahidi F. Antioxidant potential of sea cucumbers and their beneficial effects on human health. Mar Drugs [Internet]. 2022;20(8):521. Available from: https://www.mdpi.com/1660-3397/20/8/521

7. Nursid M, Marraskuranto E, Chasanah E. Cytotoxicity and apoptosis induction of sea cucumber Holothuria atra extracts. Pharmacognosy Res. 2019;11(1):41–6.

8. Grauso L, Yegdaneh A, Sharifi M, Mangoni A, Zolfaghari B, Lanzotti V. Molecular networking-based analysis of cytotoxic saponins from sea cucumber Holothuria atra. Mar Drugs [Internet]. 2019;17(2):86. Available from: https://www.mdpi.com/1660-3397/17/2/86

9. Shahinozzaman M, Ishii T, Takano R, Halim M, Hossain M, Tawata S. Cytotoxic desulfated saponin from Holothuria atra predicted to have high binding affinity to the oncogenic kinase PAK1: a combined in vitro and in silico study. Sci Pharm [Internet]. 2018;86(3):32. Available from: https://www.mdpi.com/2218-0532/86/3/32

10. Puspitasari Y, Tuenter E, Foubert K, Herawati H, Hariati A, Aulanni’am A, et al. Saponin and fatty acid profiling of the sea cucumber Holothuria atra, α-glucosidase inhibitory activity and the identification of a novel triterpene glycoside. Nutrients [Internet]. 2023;15(4):1033. Available from: https://www.mdpi.com/2072-6643/15/4/1033

11. Nursid M, Patantis G, Dewi A, Achmad M, Sembodo P, Estuningsih S. Immunnostimulatory activity of Holothuria atra sea cucumber. Pharmacia [Internet]. 2021;68:121–7. Available from: https://doi.org/10.3897/pharmacia.68.e58820

12. Dakrory A, Fahmy S, Soliman A, Mohamed A, Amer S. Protective and curative effects of the sea cucumber Holothuria atra extract against DMBA-induced hepatorenal diseases in rats. Biomed Res Int [Internet]. 2015;2015(1):563652. Available from: https://onlinelibrary.wiley.com/doi/abs/10.1155/2015/563652

13. Ikhsan I, Idroes R, Azharuddin A, Nasution R, Yusnaini R, Iqhrammullah M. Fatty acid-rich extract from Holothuria atra for hyperuricemia via expressions modulation of GLUT9a and GLUT9b in rat model. Molecules [Internet]. 2023;28(10):3981. Available from: https://www.mdpi.com/1420-3049/28/10/3981

https://www.mdpi.com/1420-3049/28/10/3981

14. Hashim R, Azizan N, Zamli Z, Zulkipli F, Mazlan N, Althunibat O. Toxicity effects of water extracts of Holothuria atra jaeger in mice. Asian Pac J Trop Biomed. 2014;4(8):614–7. https://doi.org/10.12980/APJTB.4.201414B154

15. Hanafi P, Sutrisno A, Murniasih T, Harijono, Putra M, Untari F. Acute oral toxicity assessment of the ethanol extract of Holothuria atra in mice. Asian J Pharm Clin Res. 2019;13:150–3. https://doi.org/10.22159/ajpcr.2020.v13i2.36316

16. Nursid M, Dewi A, Patantis G, Marraskuranto E, Habibullah U, Subagja M, et al. Subchronic toxicity of Holothuria atra ethanol extract for 90 days in histopathology of the liver and kidney of rats. Farmacia. 2023;71(2):322–8. https://doi.org/10.31925/farmacia.2023.2.12

17. Roepstorff A, Nansen P. Epidemiology, diagnosis and control of helminth parasites of swine. Rome, Italy: Food and Agriculture Organization of the United Nations; 1998. p 161.

18. OECD. OECD guideline for the testing of chemicals; test no. 408: repeated dose 90-day oral toxicity study in rodents [Internet]. (OECD guidelines for the testing of chemicals, Section 4). Paris, France: OECD; 2018 [cited 2024 Aug 1]. Available from: https:// www.oecd-ilibrary.org/environment/test-no-408-repeated-dose-90- day-oral-toxicity-study-in-rodents_9789264070707-en

19. Cheville NF. Introduction to veterinary pathology. 3rd ed. Hoboken, NJ: Wiley-Blackwell; 2006. 1–384 pp.

20. Iji P, Saki A, Tivey D. Body and intestinal growth of broiler chicks on a commercial starter diet. 1. Intestinal weight and mucosal development. Br Poult Sci [Internet]. 2001;42(4):505–13. doi: https://doi.org/10.1080/00071660120073151

21. Gartner L, Hiatt J. Color textbook of histology. 3rd ed. Philadelphia, PA: Elsevier Health Sciences; 2007. p 396.

22. Reece J, Urry L, Cain M, Wasserman S, Minorsky P, Jackson R. Campbell biology. 9th ed. Boston, MA: Pearson Learning Solutions; 2011. p 1464.

23. Xu RJ, Cranwell PD, editors. The neonatal pig: gastrointestinal physiology and nutrition. England: Nottingham University Press; 2003. p 1–360.

24. Engelking L. Review of veterinary physiology. Jackson, WY: Teton NewMedia; 2002. pp 1–657.

25. Parker G, Papenfuss T. Chapter 10?Immune system. In: Parker GA, Picut CA, editors. Atlas of histology of the juvenile rat [Internet]. Boston, MA: Academic Press; 2016. pp. 293–347. Available from: https://www.sciencedirect.com/science/article/pii/B9780128026823000100

26. Ermund A, Schütte A, Johansson M, Gustafsson J, Hansson G. Studies of mucus in mouse stomach, small intestine, and colon. I. Gastrointestinal mucus layers have different properties depending on location as well as over the Peyer’s patches. Am J Physiol- Gastrointest Liver Physiol [Internet]. 2013;305(5):G341–7. doi: https://doi.org/10.1152/ajpgi.00046.2013

27. Kang W, Rathinavelu S, Samuelson L, Merchant J. Interferon gamma induction of gastric mucous neck cell hypertrophy. Lab Investig. 2005;85(5):702–15.

28. Fahmy S, Amer M, Al-Killidar M. Ameliorative effect of the sea cucumber Holothuria arenicola extract against gastric ulcer in rats. J Basic Appl Zool [Internet]. 2015;72:16–25. Available from: https://www.sciencedirect.com/science/article/pii/S2090989615000223

29. Subramaniam B, Amuthan A, D’almeida P, Arunkumar H. Efficacy of gamat extract in wound healing in albino wistar rats. Int J Pharm Sci Rev Res [Internet]. 2013;20(1):142–5. Available from: https://api.semanticscholar.org/CorpusID:46960424

30. Chauhan R. Illustrated veterinary pathology: general and systematic pathology. 2nd ed. Lucknow, India: International Book Distributing Co.; 2008.

31. Jones T, Popp J, Mohr U. Digestive system [Internet]. Berlin, Germany: Springer Berlin Heidelberg; 1997. (Monographs on pathology of laboratory animals). Available from: https://books.google.co.id/books?id=Fy1tAAAAMAAJ

32. Toljamo K. Gastric erosions?clinical significance and pathology : a long-term follow-up study [Internet]. Oulu, Finland: University of Oulu; 2012. Available from: https://api.semanticscholar.org/CorpusID:43797210

33. Antonucci L, Fagman J, Kim J, Todoric J, Gukovsky I, Mackey M, et al. Basal autophagy maintains pancreatic acinar cell homeostasis and protein synthesis and prevents ER stress. Proc Natl Acad Sci U S A [Internet]. 2015;112(45):E6166–74. Available from: https://www.pnas.org/doi/abs/10.1073/pnas.1519384112

34. Shirahata S, Hamasaki T, Teruya K. Advanced research on the health benefit of reduced water. Trends Food Sci Technol [Internet]. 2012;23(2):124–31. Available from: https://www.sciencedirect.com/science/article/pii/S0924224411002408

35. Suttie A, Masson R, Schutten M. Chapter 10?Exocrine pancreas. In: Suttie AW, editor. Boorman’s pathology of the rat [Internet]. 2nd ed. Boston, MA: Academic Press; 2018. pp. 107–22. Available from: https://www.sciencedirect.com/science/article/pii/B9780123914484000101

36. Saad D, Baiomy A, Mansour A. Antiseptic effect of sea cucumber (Holothuria atra) against multi-organ failure induced by sepsis: molecular and histopathological study. Exp Ther Med. 2016;12(1):222–30. https://doi.org/10.3892/etm.2016.3321

37. Munday JS. Pathologic basis of veterinary disease [Internet]. In: Zachary JF, editor. Veterinary dermatology. 6th ed. St. Louis, MO: Elsevier; 2017. Vol. 28, p 258. Available from: https://onlinelibrary.wiley.com/doi/abs/10.1111/vde.12434

38. Bordbar S, Anwar F, Saari N. High-value components and bioactives from sea cucumbers for functional foods—a review. Mar Drugs [Internet]. 2011;9(10):1761–805. Available from: https://www.mdpi.com/1660-3397/9/10/1761

39. Damjanov I. Pathology secrets. Maryland Heights, MO: Mosby/ Elsevier; 2009. p 509.

40. Reilly J. Euthanasia of animals used for scientific purposes. Adelaide, Australia: ANZCCART; 2001. p 106.

41. Zolhavarieh S, Nourian A, Sadeghi-Nasab A. A new method for on-farm euthanasia with animal welfare considerations. Iran J Vet Surg [Internet]. 2011;6(1–2):55–64. Available from: https://www.ivsajournals.com/article_3134.html

42. Duke-Novakovski T, de Vries M, Seymour C. BSAVA manual of canine and feline anaesthesia and analgesia. 3rd ed. Chorley, UK: British Small Animal Veterinary Association; 2016. pp 1–464.

43. Buchanan N. Diet formulation and manufacturing technique interactions affect pellet quality and broiler growth [dissertation]. Morgantown, WV: West Virginia University; 2008.

44. Tufarelli V, Desantis S, Zizza S, Laudadio V. Performance, gut morphology and carcass characteristics of fattening rabbits as affected by particle size of pelleted diets. Arch Anim Nutr [Internet]. 2010;64(5):373–82. doi: https://doi.org/10.1080/1745039X.2010.496945

45. Fox J, Barthold S, Davisson M, Newcomer C, Quimby F, Smith A. The mouse in biomedical research: normative biology, husbandry, and models. 2nd ed. In: Fox JG, Barthold S, Davisson M, Newcomer CE, Quimby FW, Smith A, editors. San Diego, CA: Academic Press; 2006. Vol. 3.

46. Sherwood L. Human physiology: from cells to systems. 6th ed. Melbourne, Australia: Cengage Learning Australia; 2016.

47. Clevers H. The intestinal crypt, a prototype stem cell compartment. Cell [Internet]. 2013;154(2):274–84. Available from: https://www.sciencedirect.com/science/article/pii/S0092867413008386

48. Clevers H, Bevins C. Paneth cells: maestros of the small intestinal crypts. Annu Rev Physiol [Internet]. 2013;75:289–311. Available from: https://www.annualreviews.org/content/journals/10.1146/annurev-physiol-030212-183744

49. Olivera-Castillo L, Grant G, Kantún-Moreno N, Acevedo-Fernández J, Puc-Sosa M, Montero J, et al. Sea cucumber (Isostichopus badionotus) body-wall preparations exert anti-inflammatory activity in vivo. PharmaNutrition [Internet]. 2018;6(2):74–80. Available from: https://www.sciencedirect.com/science/article/pii/S2213434418300264

50. Wargasetia T, Ratnawati H, Widodo N, Widyananda M. Antioxidant and anti-inflammatory activity of sea cucumber (Holothuria scabra) active compounds against KEAP1 and iNOS protein. Bioinform Biol Insights [Internet]. 2023;17:11779322221149612. Available from: https://doi.org/10.1177/11779322221149613

51. Erben U, Loddenkemper C, Doerfel K, Spieckermann S, Haller D, Heimesaat M, et al. A guide to histomorphological evaluation of intestinal inflammation in mouse models. Int J Clin Exp Pathol [Internet]. 2014;7(8):4557–76. Available from: www.ijcep.com/

52. Sharp P, Villano J. The laboratory rat. 2nd ed. Boca Raton, FL: CRC Press; 2012.

53. Eurell J, Frappier B. Dellmann’s textbook of veterinary histology. 6th ed. Eurell JA, Frappier BL, editors. Ames, IA: Blackwell Publishing Professional; 2006.

54. Treuting P, Dintzis S, Frevert C, Liggitt D, Montine K. Comparative anatomy and histology: a mouse and human atlas (Expert consult). 1st ed. London, UK: Academic Press; 2012.

55. Esmat AY, Said MM, Soliman AA, El-Masry KSH, Badiea EA. Bioactive compounds, antioxidant potential, and hepatoprotective activity of sea cucumber (Holothuria atra) against thioacetamide intoxication in rats. Nutrition [Internet]. 2013;29(1):258–67. Available from: https://www.sciencedirect.com/science/article/pii/S089990071200264X

Article Metrics
28 Views 14 Downloads 42 Total

Year

Month

Related Search

By author names