Protection of Centella asiatica Extract Through BDNF Expression on Stunting Model Zebrafish Larvae ( Danio rerio ) by Rotenone Induced

Abstract

Objectives: Stunting in WHO Child Growth Standards is based on the length-forage indexes with less than-2 SD of z score. Rotenone is a pesticide model that acts as an mitochondrial complex I inhibitor and Endocrine Disrupting Chemicals (EDCs). Brain Derived Neurotrophic Factor (BDNF) is one of growth factors mostly found in central nervous system and slightly in peripheral nerves. One of natural herb, Centella asiatica, has antioxidant effects and grows in tropical areas in Indonesia. The aim of this research was to know the effects of Centella asiatica extracts on zebrafish larvae (Danio rerio) by Rotenone induced, through Brain Derived Neurotropic Factor (BDNF) expression. Methods: Zebrafish embryos were divided into five groups including control (C), rotenone group (R), and Centella asiatica group (Ca1;Ca2;Ca3) with concentration of the each 2.5, 5, and 10 g/mL. The entire groups had been started since 2 until 72 hpf (half post fertilization) and followed its development up to the age of larvae 9 dpf (days post fertilization). It was followed by measuring the body length at the age of 3, 6 and 9 dpf using Image Raster software. After termination zebrafish larvae, the BDNF expression measurement was measure by using an Immunohistochemistry (IHC) wholemount method with diamino benzidine (DAB). Brown colour density values was quantified by using Image J software. Results : There was a significant difference in the mean of body length which was shown by Ca2 group (5 g/ml) compared with rotenone group (p < 0,05). The BDNF expression increased in all three Centella asiatica groups compared with rotenone group. However, only Centella asiatica group (5 g/ml) showed the most significant effect on the increase of BDNF expression (p <0.05). Conclusion: Protection of Centella asiatica extract increases the body length and BDNF expression on the stunting model zebrafish larvae induced by rotenone.