Protection of Pegagan (Centella asiatica) Extract Through Hsp60 and Bax Expression on Stunting Model Zebrafish Larvae (Danio rerio) By Rotenone Inducted

Abstract. Objective: Rotenone is a pesticide (insecticide and fish killers) that is widely used in
water. This study utilised rotenone as a model to initiate the occurrence of stunting on
zebrafish larvae. Rotenone acts to inhibit the mitochondrial complex I to form an Adenine Tri
Phosphate (ATP), Centella asitica(CA) is commonly used as Ayurveda medication as the
triterpene in CA’s ingredient and antioxidant protect cells from oxidative damage. This study
aimed to know the effect of CA extract on the stunting model zebrafish larvae induced by
rotenone through Hsp60 and Bax expression. Methods: Zebrafish embryos were exposed with
rotenone [10 ppb] and CA extracts(concentration 2,5 μg/mL; 5 μg/mL and 10 μg/Ml) from the
age of 2 to 72 hpf (hour post fertilization).The body length was measured at the age of 3, 6, and
9 dpf (day post fertilization) using Image Raster software. The measurement of Hsp60 and Bax
expression used IHC method on wholemount zebrafish larvae through DAB colouring. Density
integrated was observed with Image J software. Results: The results of the study showed,
There was an increase in Hsp60 and Bax expression on the zebrafish larvae induced by
rotenone. In addition, CA extract with concentration [5 μg/mL] increased the body length at
the age of 9 dpf (p<0,05). The observation of Hsp60 and Bax expression showed there was a
significant decline on the CA extract groups with concentration [5 μg/mL]. Conclusion: CA
extracts with concentration [5 μg/mL] increase the body length of zebrafish larvae and
decreased Hsp60 and Bax expression.