Downregulation of liver HSP70 in zebrafish exposed to Jatropha curcas seed cake extract

• Published in: Cogent environmental science 2016-12, Vol.2 (1), p.1179563
• Main Authors: Hallare, A.V., Castillo, R.L., De Guzman, A.L.B., Balolong, M.P.B., Lazaro, J.E.H.
• Format: Article
• Language: English
• Subjects: Acute toxicity; Alternative energy sources; Biodiesel fuels; Biofuels; Danio rerio; downregulation; Exposure; Heat shock; heat shock protein 70; Heat shock proteins; Hepatocytes; Hsp70 protein; Inactivation; Jatropha curcas; Lectins; Liver; mRNA; phorbol esters; Phytochemicals; Polymerase chain reaction; Profiling; Renewable energy; Renewable energy sources; Toxicity testing; Zebrafish
• ISSN: 2331-1843; 2331-1843; 2765-8511
• DOI: 10.1080/23311843.2016.1179563

The use of Jatropha curcas in biodiesel production is one of the emerging alternative sources of renewable energy. However, the extraction process leaves a seed cake waste and the lack of disposal protocols poses serious threat to ecosystems. This study aims to determine the toxicity of the seed cake extract to Danio rerio (zebrafish) through the analysis of heat shock protein 70 expression. The median lethal concentration (LC50) was first determined through an acute toxicity test coupled with probit analysis. For the HSP70 induction test, fishes were subjected to a non-exposed setup and sublethal concentration (50% of LC50) of detoxified, washed, and untreated seed cake. Total RNA from liver samples was isolated and reverse transcribed followed by amplification through polymerase chain reaction (PCR) and gel profiling. Results indicate an LC50 of 0.172% (w/v) or 1.72 g/L. The heat shock induction test showed a significant downregulation of HSP70 expression in fishes exposed to washed and untreated setups. This underexpression may be attributed to phytochemicals such as lectins which have been widely reported to interact with HSP70. Furthermore, mRNA levels in the detoxified setup have no significant difference with the non-exposed setup, possibly indicating a successful inactivation of toxic phytochemicals in the seed cake. Moreover, the present study shows that downregulation of HSP70 is also an indicator of stress, and low HSP70 levels may lead to a decreased resistance of liver cells to stress-inducing agents.