Propolis Ability on Gene Expression Regulation of cetp, hmgcra, mtp, and cyp7a1a in Zebrafish (Danio rerio) with High Cholesterol Diet Induction

• Main Author: Sativa, Novriza
• Format: Theses
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/36267
• Institution: Institut Teknologi Bandung

Cardiovascular disease (CVD) is the leading cause of serious death with a high mortality rate in the worldwide, including in Indonesia. This disease can be caused by high levels of cholesterol which capable of causing atherosclerosis by forming plaque in blood vessels. Statins was used in several countries to control high cholesterol levels to inhibit the formation of cholesterol, but clinically side effect of this drug for long term still remain unknown. Various studies have been conducted to find out the natural product for alternative treatments in lowering cholesterol levels, one using the propolis. Propolis as a natural material has a high levels of polyphenol compounds which could be expected to inhibit the formation of plaque in blood vessels. But the use of propolis in the society has grown in the absence of preclinical testing in the laboratory to determine the treatment target and mechanism of material extracts that can treat a variety of diseases, including hypercholesterolemia. Propolis is known to able lower cholesterol levels from various studies, but the molecular mechanisms still questionable. Therefore, it is necessary to study the pathways associated with cholesterol to prevent the occurrence of acuumulation of cholesterol and plaques in blood vessels using animal models zebrafish larvae. The aim of this study was to determine the ability of propolis on relative gene expression hmgcra, mtp, cyp7a1a and cetp in zebrafish with high cholesterol diet induction. In this study the compounds or red propolis was analyzed by using GC-MS. The extract of propolis is used to treated 5dpf (days post fertilization) larval zebrafish for 7 days after feeding high cholesterol diet (HCD) every day for 3 hours. HCD iv additional foods containing 4% (w / w) cholesterol accompanied by the BODIPY 0.002% (w / w) as fluorescent marker cholesteryl ester in each group. Concentration 1 ppm of propolis from preliminary study with serial concentrations was used in group treatment. Positive control group was treated with ezetimibe as a safe drug in lowering hypercholesterolemia for use in larval zebrafish, then molecular analysis of gene cetp, hmgcra, mtp and cyp7a1a performed by qPCR. The results show that red propolis from bee Trigona sp. have high levels of polyphenols which is equal to 58.11%. Fluorescence microscope observation showed that induction with high cholesterol diet can formed accumulation of cholesterol in blood vessels larval zebrafish. Administration of propolis on larval zebrafish is able to prevent the formation of accumulation of cholesterol. This data is supported by the relative gene expression of cetp (as a marker gene of hypercholesterolemia), mtp (a lipid transfer protein required for assembly and secretion of lipoprotein), and cyp7a1a (a rate limiting enzyme for bile acid biosynthesis) are decreasing, while hmgcra (a rate limiting enzyme in cholesterol biosynthesis) increased. This shows that the administration of propolis is able to prevent the accumulation of cholesterol. From this study can be concluded that the polyphenol compounds from propolis are able to prevent cholesterol accumulation in blood vessels. Down regulation mechanism on cetp and mtp genes which give hypocholesterolemic effects is thought to be caused by polyphenol. The results from this study, is expected to be applied propolis in humans to prevent and control the conditions of hypercholesterolemia. The development of further research is expected to test variations of propolis dosage to humans.