THE ABILITY OF ETHANOLIC EXTRACT OF PROPOLIS (EEP) TO INDUCE CAUDAL FIN REGENERATION IN HYPERGLYCEMIC ZEBRAFISH (Danio rerio) THROUGH CANONICAL WNT SIGNALING PATHWAY

• Main Author: Devi Indriani, Annisa
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/34187
• Institution: Institut Teknologi Bandung

Diabetes mellitus (DM) is a complex metabolic syndrome disorder with a high mortality rate in the world, and characterized by hyperglycemia condition. Impaired wound healing is one of the long-term complications caused by DM. Hyperglycemia causes a decrease in regeneration which is known to be associated with decreased proliferation. The canonical Wnt signaling pathway is one of the pathways that playing roles in the proliferation, migration and differentiation of cells known to be involved in delay of wound healing. One of the approaches taken in the development of diabetes drugs through the canonical Wnt signaling cascade is the use of phytochemical compounds as naturally based alternative therapies. The Ethanolic extracts of propolis (EEP) with its phenolic compounds is known to have a high antioxidant activity which is supposed to lower blood glucose levels and enhance tissue regeneration along with increased expression of ?-catenin and LEF-1 genes. in Wnt signaling cascade in the regeneration process. The purpose of this study were to determine the effect of EEP towards the blood glucose levels of hyperglycemic zebrafish (Danio rerio) induced by alloxan, to study EEP effect on the caudal fins of hyperglycemic zebrafish regeneration, as well as to study the effect of EEP on gene expression of ?-catenin gene and LEF-1 gene at the mRNA level as reviewed through canonical Wnt signaling cascade. This study used seven treatment groups as following: control (K), control of propylene glycol (KPG 1%) as the solvent, control of propolis (KP 15 ppm), hyperglycemia (P1), hyperglycemia + propylene glycol 1% (P2), hyperglycemia + EEP 15 ppm (P3), hyperglycemia + metformin 10 mL (P4), with 20 zebrafish in each treatment group. Four treatment groups with the induction of hyperglycemia were treated by exposing or allowing 20 zebrafish (10 zebrafish / aquarium) to swim in a solution of 400 mg of alloxan/ 100 ml NaCl 0,45% with the duration of 1 hour per day for 5 days. Then, the fish were transferred into 2 L of 2% glucose solution with the duration of 24 hours per day for 5 days (the replacement of glucose solution performed once every 24 hours). Afterwards, the caudal fins were cut after the induction hyperglycemia on the first iv day, followed by exposing or allowing the zebrafish of each groups to swim in 1% PG solution, 15 ppm EEP solution, and a 10 mL solution of metformin for 4 days after amputation until the caudal fin regeneration occurred. Blood glucose level of 5 zebrafish of each group was measured using glucometer in the forth day after amputation. Quantification of the caudal fin regeneration was conducted by using Image J software. The blood glucose levels and caudal fins regeneration in zebrafish were analyzed using normality test and one-way ANOVA to determine the significant difference between the treatment groups. Meanwhile, the analysis of ?-catenin gene expression profiles and LEF-1 mRNA levels were performed using RT-qPCR. The result of this study showed that the concentration of 15 ppm EEP was able to lower blood glucose levels significantly (P<0.05) from 339,4 mg/dL to 243,2 mg/dL in hyperglicemic zebrafish. However it had not reached normal level of blood glucose yet. The 15 ppm EEP was also able to increase the percentage of the caudal fin blastema regeneration significantly (P<0.05) and increase the expression of ?-catenin and LEF-1 genes. Based on the result of this study, it concluded that the concentration of 15 ppm EEP was able to lower hiperglicemia levels although it had not reached normal level of blood glucose yet for 4 days. However, the concentration of the EEP was able to increase the caudal fin regeneration and the relative expression of genes ?-catenin and LEF-1 in hyperglycemic zebrafish.