Acute Toxicity of Trypsin Inhibitor from Tamarind Seeds in Embryo and Adult Zebrafish ()

Overview

Journal Toxicol Rep

Date 2024 Oct 29

PMID 39469098

Authors

Yohanna Layssa Dos Santos Melo

Ana Carolina Luchiari

Beatriz Silva Lopes

Maria Gabriela Ferreira Rocha Silva

Tatiana Dos Santos Pais

Joao Eduardo Procopio Gama Cortez

Christina da Silva Camillo

Sergio Adriane Bezerra de Moura

Juliana Kelly da Silva-Maia

Ana Heloneida de Araujo Morais

Affiliations

Soon will be listed here.

Abstract

The trypsin inhibitor isolated from tamarind seeds (TTI) is being investigated for potential applications in the treatment of noncommunicable diseases (NCD), such as hypertension, obesity, and diabetes. This study aimed to assess TTI embryotoxicity and acute toxicity in adult zebrafish (). TTI was extracted and isolated from tamarind seeds. Embryonic and adult zebrafish were exposed for 96 hours to three concentrations of TTI (12.5, 25, and 50 mg/L). Zebrafish embryos (n=60 per group) were evaluated for survival, hatching, malformations, and potential developmental marker alterations, in addition to cardiotoxicity and neurotoxicity tests. For acute toxicity assessment in adults (n=20 per group), survival and locomotor and anxiety-like behaviors were assessed, along with genotoxicity (micronucleus) evaluation. Embryos exposed to TTI showed no significant adverse effects, presented normal heart rates and positive reflex response in the neurotoxicity tests. In adult fish, TTI did not cause mortality or significant behavioral changes, suggesting no neurotoxicity and no genotoxicity. Histopathological analyses of the whole body showed only changes in the liver and spinal cord, similar to those observed in the control group not exposed to TTI. These findings indicate TTI's biosafety and therapeutic potential in complex organisms. Further research is required to evaluate its long-term effects and efficacy in treating non-communicable diseases.

References

Dos Santos Pais T, Luchiari A, Souza A, Medeiros I, Ferreira Rocha Silva M, Dos Santos Y . Assessment of acute toxicity of crude extract rich in carotenoids from Cantaloupe melon (Cucumis melo L.) and the gelatin-based nanoparticles using the zebrafish (Danio rerio) model. Food Chem Toxicol. 2023; 181:114091. DOI: 10.1016/j.fct.2023.114091. View

Zang L, Maddison L, Chen W . Zebrafish as a Model for Obesity and Diabetes. Front Cell Dev Biol. 2018; 6:91. PMC: 6110173. DOI: 10.3389/fcell.2018.00091. View

Ghaddar B, Diotel N . Zebrafish: A New Promise to Study the Impact of Metabolic Disorders on the Brain. Int J Mol Sci. 2022; 23(10). PMC: 9141892. DOI: 10.3390/ijms23105372. View

Linney E, Upchurch L, Donerly S . Zebrafish as a neurotoxicological model. Neurotoxicol Teratol. 2004; 26(6):709-18. DOI: 10.1016/j.ntt.2004.06.015. View

McMullen P, Andersen M, Cholewa B, Clewell 3rd H, Dunnick K, Hartman J . Evaluating opportunities for advancing the use of alternative methods in risk assessment through the development of fit-for-purpose in vitro assays. Toxicol In Vitro. 2018; 48:310-317. DOI: 10.1016/j.tiv.2018.01.027. View

de Medeiros A, de Souza B, Pinheiro Coutinho L, Murad A, Dos Santos P, Monteiro N . Structural insights and molecular dynamics into the inhibitory mechanism of a Kunitz-type trypsin inhibitor from L. J Enzyme Inhib Med Chem. 2021; 36(1):480-490. PMC: 7875565. DOI: 10.1080/14756366.2021.1876686. View

Li S, Liu L, He G, Wu J . Molecular targets and mechanisms of bioactive peptides against metabolic syndromes. Food Funct. 2017; 9(1):42-52. DOI: 10.1039/c7fo01323j. View

de Siqueira Patriota L, do Nascimento Santos D, Barros B, de Souza Aguiar L, Silva Y, Dos Santos A . Evaluation of the In Vivo Acute Toxicity and In Vitro Hemolytic and Immunomodulatory Activities of the Moringa oleifera Flower Trypsin Inhibitor (MoFTI). Protein Pept Lett. 2020; 28(6):665-674. DOI: 10.2174/0929866527999201113105858. View

Ribeiro J, Serquiz A, Silva P, Barbosa P, Sampaio T, de Araujo Junior R . Trypsin inhibitor from tamarindus indica L. seeds reduces weight gain and food consumption and increases plasmatic cholecystokinin levels. Clinics (Sao Paulo). 2015; 70(2):136-43. PMC: 4351314. DOI: 10.6061/clinics/2015(02)11. View

10.

Jegadheeshwari S, Velayutham M, Gunasekaran K, Kesavan M . DbGTi: Thermostable trypsin inhibitor from Dioscorea bulbifera L. ground tubers: assessment of antioxidant and antibacterial properties and cytotoxicity evaluation using zebrafish model. Int J Biol Macromol. 2024; 263(Pt 2):130244. DOI: 10.1016/j.ijbiomac.2024.130244. View

11.

de Oliveira G, Nascimento A, Luz A, Aguiar A, Lima M, Matias L . Prospecting antibacterial activity of a peptide from trypsin inhibitor isolated from tamarind seed. J Enzyme Inhib Med Chem. 2022; 38(1):67-83. PMC: 9621272. DOI: 10.1080/14756366.2022.2134997. View

12.

Johnson A, Loh E, Verbitsky R, Slessor J, Franczak B, Schalomon M . Examining behavioural test sensitivity and locomotor proxies of anxiety-like behaviour in zebrafish. Sci Rep. 2023; 13(1):3768. PMC: 9992706. DOI: 10.1038/s41598-023-29668-9. View

13.

Teixeira J, Souza A, Macedo-Sampaio J, Menezes F, Pereira B, Batistuzzo de Medeiros S . Embryotoxic Effects of Pesticides in Zebrafish (): Diflubenzuron, Pyriproxyfen, and Its Mixtures. Toxics. 2024; 12(2). PMC: 10892354. DOI: 10.3390/toxics12020160. View

14.

Maximino C, de Brito T, Colmanetti R, Pontes A, de Castro H, de Lacerda R . Parametric analyses of anxiety in zebrafish scototaxis. Behav Brain Res. 2010; 210(1):1-7. DOI: 10.1016/j.bbr.2010.01.031. View

15.

Aguiar A, Queiroz J, Santos P, Camillo C, Serquiz A, Costa I . Beneficial Effects of Tamarind Trypsin Inhibitor in Chitosan-Whey Protein Nanoparticles on Hepatic Injury Induced High Glycemic Index Diet: A Preclinical Study. Int J Mol Sci. 2021; 22(18). PMC: 8470918. DOI: 10.3390/ijms22189968. View

16.

Rhea E, Banks W . Role of the Blood-Brain Barrier in Central Nervous System Insulin Resistance. Front Neurosci. 2019; 13:521. PMC: 6558081. DOI: 10.3389/fnins.2019.00521. View

17.

Parasuraman S . Toxicological screening. J Pharmacol Pharmacother. 2011; 2(2):74-9. PMC: 3127354. DOI: 10.4103/0976-500X.81895. View

18.

Costa I, Medeiros A, Carvalho F, Lima V, Serquiz R, Serquiz A . Satietogenic Protein from Tamarind Seeds Decreases Food Intake, Leptin Plasma and CCK-1r Gene Expression in Obese Wistar Rats. Obes Facts. 2018; 11(6):440-453. PMC: 6341364. DOI: 10.1159/000492733. View

19.

Chang C, Li H, Zhang R . Zebrafish facilitate non-alcoholic fatty liver disease research: Tools, models and applications. Liver Int. 2023; 43(7):1385-1398. DOI: 10.1111/liv.15601. View

20.

Carvalho F, Lima V, Costa I, Luz A, Ladd F, Serquiz A . Anti-TNF-α Agent Tamarind Kunitz Trypsin Inhibitor Improves Lipid Profile of Wistar Rats Presenting Dyslipidemia and Diet-induced Obesity Regardless of PPAR-γ Induction. Nutrients. 2019; 11(3). PMC: 6470745. DOI: 10.3390/nu11030512. View