Assessing Antioxidant Enzyme Activity in Zebrafish Larvae Treated with Diosgenin and Doxorubicin

Abstract

In this research, zebrafish larvae models were employed to evaluate the activity of antioxidant enzymes—GSH, GPX, and GST—in response to treatments with doxorubicin and a novel drug, diosgenin. Solutions for the two exposure groups, Doxorubicin (30 µM) and diosgenin (37.8 µM), were prepared by diluting stock solutions in egg water. The zebrafish maintenance and egg collection process involved housing breeding groups in a specific spawning tank with a 1:1 male-to-female ratio. The tank was equipped with a collection box and mesh at the bottom to gather embryos and prevent consumption by adult fish. The study comprised two groups, each with 15 samples, totaling 30 samples. Group 1 assessed doxorubicin’s effects on larval zebrafish, while Group 2 evaluated diosgenin’s impact. Embryos, with n=15 per plate, were sorted into petri plates for each exposure group and exposed to diosgenin from 4 hpf to 96 hpf (Hours Post Fertilisation). All conditions aligned with the OECD guidelines for fish embryo toxicity assays, ensuring the study’s validation. Every experiment was conducted in triplicate. Statistical analysis utilised the Statistical Packages for the Social Sciences (SPSS) software. The analysis maintained a confidence ratio of 95%, a threshold of 0.01, an 80% G power, and an enrolment ratio of 1. The findings revealed that zebrafish larvae treated with the novel drug, diosgenin, exhibited heightened levels of GPx, GSH, and GST compared to the doxorubicin-treated group. This increase was statistically significant with a p-value of 0.000 (p<0.05). Consequently, this study demonstrated that diosgenin and doxorubicin treatments upregulated antioxidant enzyme activity—GPX, GSH, and GST—in zebrafish larvae, highlighting diosgenin as an effective antioxidant agent.