Comparative Estimation of ROS Levels using DCFDA in Zebrafish Larvae Model on the Treatment of Camphene and Doxorubicin for Antioxidant Property

Department of Biotechnology, Saveetha School of Engineering, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, Tamil Nadu, India. Pincode : 602105.

^* Corresponding author: prakashkalpana067@gmail.com

Abstract

To assess the impact of Camphene and Doxorubicin on ROS levels in Zebrafish larvae, the study induced oxidative stress using 1 mM H2O2. DCFDA enzymatic assay was utilised to detect intracellular ROS levels in the zebrafish larvae. The compounds were prepared into a solution by diluting the stock solution with DMSO. Zebrafish were bred in a spawning tank at a 1:1 female-to-male ratio, and their eggs were collected, segregated into four petri plates, and exposed to Camphene from 4 hours post fertilization (hpf) until 96hpf. The hatching and mortality rates were monitored at 96hpf, calculated as the ratio of hatched larvae to the total number of live larvae at observation. Statistical analysis was conducted using SPSS software with preset parameters (threshold of 0.05, 80% G power, sample size n=15, 95% confidence interval, and an enrolment ratio of 1). The study revealed significant differences (p=0.000, p<0.05) between the compound-exposed embryos (62.4667±1.88) and the control group (34.4000±0.07). Early exposure to Camphene at 26 μmol demonstrated a positive impact on the antioxidant capacity in zebrafish larvae, as indicated by fluorescence intensity. This suggests that Camphene treatment effectively counteracts H2O2-induced ROS production in zebrafish larvae. Specifically, fluorescence intensity was 30% lower in the Camphene-exposed group compared to the Doxorubicin-exposed group.