PENGARUH PANGAN FUNGSIONAL BLACK RICE CRUNCH TERHADAP TIKUS PUTIH (Rattus norvegicus Berkenhout, 1769) OBESITAS: KAJIAN HISTOPATOLOGIS HATI DAN PANKREAS, EKSPRESI RELATIF mRNA IL-1BETA PANKREAS

Chesa Ekani Maharesi, Dr. Ardaning Nuriliani, S.Si.,M.Kes ; Prof. Dra. Rarastoeti Pratiwi, M.Sc., Ph.D

2024 | Tesis | S2 Biologi

Obesitas merupakan suatu kondisi akumulasi lemak yang berlebihan dan tidak normal. Obesitas mengalami prevalensi yang signifikan di dunia. Obesitas menjadi masalah kesehatan global karena meningkatkan resiko penyakit kronis lain. Pencegahan dan pengobatan obesitas dapat menggunakan pangan fungsional. Beras hitam memiliki kandungan antosianin tinggi yang berpotensi menjadi pangan fungsional. Penelitian ini adalah untuk mengidentifikasi pengaruh Black Rice Crunch terhadap histopatologi hati dan pankreas serta ekspresi mRNA IL-1beta tikus (Rattus norvegicus Berkenhout, 1769) obesitas. Penelitian ini menggunakan tikus wistar berumur 3 bulan yang dikelompokkan menjadi 5 yaitu kontrol normal (NO), pakan obesitas (BRC 0),  pakan BRC 25% (BRC 1), pakan BRC 50% (BRC 2), dan pakan BRC 75% (BRC 3). Tikus diberi perlakuan pakan obesitas dan pakan BRC selama 8 minggu. Setelah perlakuan terakhir, di timbang berat badan tikus, tikus dikorbankan, organ hati dan pankreas dikoleksi, dan timbang berat organ hati dan pankreas. Hati dan pankreas dilakukan penyimpanan sediaan histologi dengan fiksasi menggunakan Neutral Buffer Formalin (NBF) 1%  dibuat preparat histologis menggunakan metode parafin dengan pewarnaan hematoksilin-eosin. Pankreas untuk pengukuran ekspresi mRNA IL-1beta  disimpan dalam RNA later. Parameter yang diamati pada hati yaitu nekrosis, perlemakan, degenerasi sel, arsitektur parenkim, infiltrasi leukosit. Parameter yang diamati pada pankreas yaitu cloudy swelling, degenerasi sel, fibrosis, nekrosis. Kerusakan arsitektur, dan vakuolisasi. Parameter yang diamati pada ekspresi mRNA IL-1beta   yaitu perbedaan nilai quantification cycle (cq) dari proses Qrt-PCR mRNA gen target. Metode kualitatif pada hati dan pankreas dilakukan dengan analisis deskripsi komparatif menggunakan metode binomial scoring system. Metode kuantitatif terkait pengukuran indeks organ, parameter kerusakan hati dan pankreas dilakukan dengan analisis statistik satu arah (One-Way) ANOVA serta uji Duncan (pmenggunakan software SPSS. Analisis data hasil ekspresi mRNA IL-1beta yaitu quantification cycle (Cq) dari qPCR menggunakan metode Livak dan analisis data ANOVA dengan signifikansi 5 %  IL-1beta pada pankreas tikus obesitas.

Obesity is a condition of excessive and abnormal fat accumulation. Obesity has a significant prevalence in the world. Obesity is a global health problem because it increases the risk of other chronic diseases. Prevention and treatment of obesity can use functional foods. Black rice has a high anthocyanin content which has the potential to be a functional food. This study was to identify the effect of Black Rice Crunch on liver and pancreas histopathology as well as IL-1beta mRNA expression in obese rat (Rattus norvegicus Berkenhout, 1769). This study used 3 month old Wistar rats which were grouped into 5, namely normal control (NO), obese feed (BRC 0), BRC 25% (BRC 1), BRC 50% (BRC 2), and BRC 75% ( BRC 3). Rat were treated with obesity food and BRC food for 8 weeks. After the final treatment, the rat’s body weight was weighed, the rat were sacrificed, the liver and pancreas were collected, and the weight of the liver and pancreas were weighed. The liver and pancreas were stored in histological preparations with fixation using 1% Neutral Buffer Formalin (NBF) and histological preparations were made using the paraffin method with hematoxylin-eosin staining. Pancreas for measuring IL-1beta mRNA expression was stored in RNA later. The parameters observed in the liver are necrosis, fat, cell degeneration, parenchymal architecture, leukocyte infiltration. The parameters observed in the pancreas are cloudy swelling, cell degeneration, fibrosis, necrosis. Architectural damage, and vacuolization. The parameters observed in IL-1beta mRNA expression were differences in quantification cycle (cq) values from the qPCR process of target gene mRNA. The qualitative method on the liver and pancreas was carried out by comparative descriptive analysis using the binomial scoring system method. Quantitative methods related to measuring organ indices, liver and pancreatic damage parameters were carried out using one-way ANOVA statistical analysis and Duncan’s test (pIL-1? mRNA expression results, namely quantification cycle (Cq) from qPCR using the Livak method and ANOVA (p = 5%) The results of this study show that administering BRC at a dose of 25% is most effective in improving the histological structure of the liver in terms of central vein diameter parameters. Giving BRC at a dose of 50% was most effective in improving the histological structure of the liver in terms of liver damage parameters in obese mice. Giving BRC at a dose of 50% was most effective in improving the histological structure of the pancreas as seen in the parameters of pancreatic damage in obese mice. Giving a 50% dose of BRC was most effective in reducing IL-1beta mRNA expression levels in the pancreas of obese mice.

Kata Kunci : Obesitas, Black Rice Crunch, Histopatologi Hati dan Pankreas, Ekspresi mRNA IL-1Beta