Data of multiple different high-fat diets
Creators: name: Mustika, Syifa
name: Santosaningsih, Dewi
name: Handayani, Dian
name: Rudijanto, Achmad
Publisher: name: Zenodo
Year: 2023
Subjects: normal diet
Rattus norvegicus
metabolism
inflammatory
dysbiosis
liver histology
non-alcoholic steatohepatitis
high fat diet
western diet
high fat high fructose diet

description: <strong>Background and Objectives:</strong> Non-alcoholic steatohepatitis (NASH) is a spectrum of non-alcoholic fatty liver disease (NAFLD) that can progress to advanced liver disease. Several types of diets are known to play important roles in the development of NAFLD/NASH. This study aims to determine the most appropriate composition of diet to induced NASH in animal models. <strong>Methods:</strong> This research used <em>Rattus</em> <em>norvegicus</em> strain Wistar (n = 27) which were divided into four groups and given each diet for 12 weeks: normal diet (ND, n=7), high-fat diet (HFD, n=6), western diet (WD, n=7), high-fat-high-fructose diet (HFHFD, n=7). Subjects were documented for body weight. Blood samples were taken for biochemical analysis: <em>low-density lipoprotein</em> (LDL), triglyceride, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), hepatic lipase, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and lipopolysaccharide (LPS). Feces were taken for <em>short-chain fatty acids</em> (SCFAs) analysis. Liver histology was analyzed using NAS (NAFLD activity score). The comparison test was carried out using one-way ANOVA or Kruskal Wallis test. <strong>Results: </strong>The highest mean of body weight was in the WD group (346.14 g). Liver enzymes, LDL, triglyceride, propionic acid, and acetic acid in each group were not significantly different. TNF-α, IL-6, and hepatic lipase were significant (p = 0.000; p = 0.000; p = 0.004) and the highest was in the HFD group. Butyrate level was significant (p = 0.021) and the least was in the HFHFD group (4.77 mMol/g). Only WD and HFHFD had NAS ≥ 5 (14% and 14%). The highest percent of borderline NAS was found in WD (57%). <strong>Conclusions:</strong><strong> </strong>The HFD group induced significant liver inflammation but did not produce NASH histologically, whereas the WD and HFHFD group had the potential to develop NASH because it affected metabolic and inflammatory parameters as well as liver histology.
description: Software that is required to open the data files is SPSS.
description: All procedures were reviewed and approved by Ethical Committee in Faculty of Medicine, Universitas Brawijaya (No. 66/EC/KEPK/02/2021). Twenty seven male Wistar rats with 8–12 weeks of age were obtained from Universitas Gadjah Mada. Wistar rats were kept in a humidity and temperature control room and ad libitum access of food and water. Wistar rats were randomly assigned into 4 groups: normal diet (67% carbohydrate, 21% protein, 7% fat, 5% fiber); high-fat diet (67.1% carbohydrate, 16.5% fat, 16.4% protein), western diet (52% carbohydrate, 16.1% protein, 31.7% fat), and high-fat high fructose diet (41.5% carbohydrate, 10.3% fat, 10.2% protein, 38% fructose). All diet were given for 12 weeks. All four groups were euthanized with ketamine-xylazine intravenously at the same day. All of the livers were removed and weighed. Livers were collected and stored at -20°C or fixed in 10% paraformaldehyde for further analysis. The extraction of faecal SCFAs samples was performed following the protocols described by a previous study. A total of 0.5 gr faecal samples were used, and a dilution ratio of 1:4 between samples and sterilized water was established. We subsequently vortexed the samples for 1 min and centrifuged the homogenate at 4,800 g for 15min at 4°C. Filtration of SCFA-containing supernatant was completed by using cellulose acetate membrane (GyroDisc, Orange Scientific, CA, US) and stored at -80°C until further analysis was performed. Rat serum was used to analyze biochemical parameters in Clinical Pathology Laboratory, Universitas Brawijaya , Indonesia (12). Serum ALT, AST, and ALP were performed chemically using colorimetric analysis (ADVIA 2400 Clinical Chemistry System (Siemens, Germany). Serum TNF-α , IL-6, LPS were analyzed with sandwich enzyme-linked immunosorbent assay (ELISA) method (13). Rat feces were collected from the colon, 0.5 gram for each to analyze SCFA and stored at -25°C before measurements. SCFA analysis was performed with gas chromatography method in Agricultural Techonology Laboratory at Gadjahmada University, Indonesia (14). The livers were sliced and fixed with 10% buffered formalin, embedded in paraffin, cut at a thickness of 5μm, and then stained with hematoxylin-eosin (HE) staining (12). Sample preparation was performed in the anatomical pathology laboratory in Brawijaya University, Indonesia. Liver histopathology was used to find NAFLD Activity Score (NAS). Three parameters (steatosis score 0-3; lobules inflammation score 0-3; ballooning score 0-2) were used to know NAFLD staging. Scores of 0–2 is defined non-NASH, scores of 3–4 is defined borderline, while scores ≥ 5 is considered diagnostic of NASH (15). Data are presented as the mean ± standard deviation and were analyzed with SPSS 25.0 for Windows. One-way ANOVA was carried out when the data was normally distributed and then continued with the Tukey HSD post hoc test if the data were significant. The Kruskal-Wallis test was used when the data distribution was not normal. If the results are significant then continued with the Mann-Whitney test. Data were considered significant when p &lt; 0.05.