The effect of ethanol extract of pegagan (Centella asiatica) on bone ossification and osteoclastogenesis on the stunting model of Zebrafish (Danio rerio) larvae induced by rotenone

Abstract

Stunting, according to the WHO, is defined as body length or body height with respect to the same age that is less than minus two standard deviations (<-2SD) from the median standard of the WHO for children growth. Rotenone is a pesticide that can induce an occurrence of disrupted growth on the larvae of zebrafish. Centella asiatica or pegagan is a plant that has antioxidant effects. Zebrafish embryos were treated with pegagan at concentrations of 1.25, 2.5, and 5 µg/ml. Exposure to treatments began from 2 to 72 hpf and development was monitored until the larval age of 9 dpf. Rotenone at a concentration of 12.5 ppb can induce stunting. There was a decrease in the process of bone ossification and an increase in the expression of RANKL in zebrafish larvae induced by rotenone. Meanwhile, the extract of Centella asiatica at concentrations of 2.5 µg/ml and 5 µg/ml was able to increase body length. Pegagan 5 µg/m shown the most significant effect for the increase in bone ossification (p <0.05). Observations of RANKL expression indicated significant reductions in the Centella asiatica extract treatment groups from the lowest concentration 1.25 µg/ml, up to 2.5 µg/ml and 5 µg/ml. Centella asiatica extract of 2.5 µg/ml and 5 µg/ml concentrations can increase the body length of zebrafish larvae, increase bone ossification, and lower the expression of RANKL, which has an effect on the process of osteoclastogenesis.