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Rotenone, a natural pesticide inhibit mitochondrial complex I enzyme that caused the increase of reactive oxygen species (ROS), reduced ATP production and stimulate apoptosis. This condition will affect the proliferation and development. IGF-1 (Insulin-Like Growth Factor-1) is a protein that responsible for growth and development in prenatal and postnatal of somatic cells. IRS (insulin receptor substrate) responsible for signal transduction of IGF-1 for cell differentiation, proliferation, and anti-apoptosis. Gotu kola (Centella asiatica) had long been used as herbal medicine. Its contains triterpenes as antioxidant and anti-inflammatory properties, macro and micronutrients as well. This study observed the effect of Centella asiatica (CA) extract to the expression of IGF-1, IRS and the linear growth of rotenone-induced zebrafish larvae. The zebrafish embryos were divided into 5 groups: control, Rotenone 12 ppb, Rotenone and CA extract concentration 2.5, 5 and 10 μg/ml respectively exposed from 2-hours post fertilization (hpf) until 3-days post fertilization (dpf). The expression of IGF-1 and IRS were conducted at 9 dpf by immunohistochemistry whole-mount and quantify the expressions by Image J software. The body length measured at 3, 6 and 9 dpf using Image Raster from optilab v2.2.1. The data analyzed using one way anova from SPSS v.18. The results showed that rotenone significantly decreased the IGF-1 and IRS expression (p<0.05). The administration of CA extract could be increased the expression of IGF-1 and IRS in rotenone-induced zebrafish larvae. The measurement of body length showed that rotenone reduced the body leng of larvae after 6 dpf, and 5 µg/mL of CA extract significantly increased the body length at 6 and 9 dpf. It can be concluded that CA extract increased the body length of zebrafish larvae through the increasing of IGF-1 and IRS signaling.