scholarly journals Effects of Diet Containing Soy Protein Isolate on Liver Metabolic Methylation Status Using Obese Zucker Rat Model (P08-033-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Reza Hakkak ◽  
Soheila Korourian ◽  
Oleksandra Pavliv ◽  
Stepan Melnyk
Keyword(s):  
Rat Model ◽  
Soy Protein ◽  
Methylation Status ◽  
Liver Steatosis ◽  
Soy Protein Isolate ◽  
The United States ◽  
Protein Isolate ◽  
Zucker Rats ◽  
Zucker Rat ◽  
Obese Zucker Rat

Abstract Objectives The obesity epidemic is continuing to grow in the United States and world for past two decades. There is a link between obesity and chronic diseases development such as diabetes, cardiovascular disease, certain types of cancer and liver diseases. Previously, we reported that obesity caused a significant increase liver steatosis and feeding soy protein isolate (SPI) reduced liver steatosis. The mechanism of SPI protection against liver steatosis is less known. We hypothesize that soy protein diet will reduce development of liver steatosis caused by obesity in part by changing methylation status. The objective of the present study was to investigate the effects of SPI feeding on liver metabolic methylation status using obese zucker rat model. Methods After one week of acclimation, five weeks old female lean and obese Zucker rats (n = 8/group) were randomly fed AIN-93-G diet with either casein (CAS as control) or SPI as source of protein for 22 weeks. Rats were weighted twice per week. Liver sample metabolites concentrations were measured using HPLC with Electrochemical Detection and LC-MS. Results Our results shows that; 1) obesity increased body weight significantly (P < 0.001) for both CAS and SPI diets; 2) Obese SPI-fed rats significantly (P < 0.001) reduced liver steatosis compared to obese CAS-fed rats. Also, our results show that liver metabolic profile in lean SPI-fed rats significantly (P < 0.025) increased SAM/SAH ratio (methylation ratio) compare to CAS-fed rats. Obese SPI-fed rats significantly (P < 0.001) decrease level of Homocysteine in liver and increase significantly (P < 0.001) Methionine/Homocysteine ratio. Conclusions In summary we showed that SPI diet can reduce liver steatosis by changing methylation status and improved metabolism of Homocysteine, toxic intracellular compound, through remethylation to Methionine. Funding Sources Arkansas Children's Research Institute's University Medical Group Fund grant program and Arkansas.

scholarly journals Liver Proteomics Analysis After Short- and Long-Term Soy Protein Isolate Feeding Using Obese Zucker Rat Model

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 1226-1226
Author(s):  
Melisa Kozaczek ◽  
Walter Bottje ◽  
Reza Hakkak
Keyword(s):  
Rat Model ◽  
Soy Protein ◽  
Liver Steatosis ◽  
Soy Protein Isolate ◽  
Protein Isolate ◽  
Zucker Rats ◽  
P Value ◽  
Proteomics Data ◽  
Proteomics Analysis ◽  
Z Scores

Abstract Objectives To identify possible mechanisms involved in the development and progression of NAFLD through protein expression (shotgun proteomics) analysis on liver samples of obese Zucker rats fed with either casein (CAS) or soy protein isolate (SPI) during 8 and 16 weeks. Methods 7 weeks old rats (n = 8–9 per group) were randomly assigned to an either CAS-based or a SPI-based diet. Rats were sacrificed after 8 weeks or 16 weeks of SPI feeding. Livers were immediately obtained and stored at −80 C. Ingenuity Pathway Analysis (IPA) software was used to facilitate interpretation of proteomics data. Predictions of activation or inhibition of molecules in the data was made based on activation z-score and P value of overlap (P &lt; 0.05). Activation z-scores &gt; 2.0 indicate that a molecule is activated, whereas activation z-scores of &lt; −2.0 indicate that a target molecule is inhibited. Results Upstream regulator analysis by IPA revealed 6 molecules predicted to be activated (z-scores between 2 and 2.8) and 9 inhibited (z-scores between −2 and −2.6) in SPI vs CAS-fed rats in proteomics data at 8 weeks of SPI feeding. In contrast, at 16 weeks of SPI feeding there were 12 molecules activated (z-scores between 2 and 2.5) and 18 inhibited (z-scores between −2 and −2.8) in SPI vs CAS-fed. All p values were &lt;0.05. Regulator effects analysis also revealed that some of these molecules would be participating, directly or indirectly, in the inhibition of the immune response of cells (such as IL27 and CSF2) and synthesis of lipids (CEBPA, Ins1 and IRF8) in SPI-fed rats. Conclusions These molecules and their downstream target proteins may provide clues by which soy protein produces the observed attenuation of liver steatosis that can be tested in future experiments in this obese rat model. Funding Sources This study was supported in part by the College of Medicine's University Medical Group (RH) and the Arkansas Biosciences Institute (WB, RH).

scholarly journals Long-Term Soy Protein Isolate Consumption Reduces Liver Steatosis Through Changes in Global Transcriptomics in Obese Zucker Rats

2020 ◽  
Vol 7 ◽  
Author(s):  
Melisa Kozaczek ◽  
Walter Bottje ◽  
Byungwhi Kong ◽  
Sami Dridi ◽  
Diyana Albataineh ◽  
...  
Keyword(s):  
Regression Analysis ◽  
Soy Protein ◽  
Liver Steatosis ◽  
Soy Protein Isolate ◽  
Protein Isolate ◽  
Differentially Expressed ◽  
Zucker Rats ◽  
Obese Zucker Rats ◽  
Upstream Regulators

To determine how soy protein isolate (SPI) ameliorated liver steatosis in male obese Zucker rats, we conducted global transcriptomic expression (RNAseq) analysis on liver samples of male rats fed either the SPI or a control casein (CAS)-based diet (n = 8 per group) for 16 weeks. Liver transcriptomics were analyzed using an Ilumina HiSeq system with 2 × 100 base pair paired-end reads method. Bioinformatics was conducted using Ingenuity Pathway Analysis (IPA) software (Qiagen, CA) with P &lt; 0.05 and 1.3-fold differential expression cutoff values. Regression analysis between RNAseq data and targeted mRNA expression analysis of 12 top differentially expressed genes (from the IPA program) using quantitative PCR (qPCR) revealed a significant regression analysis (r2 = 0.69, P = 0.0008). In addition, all qPCR values had qualitatively similar direction of up- or down-regulation compared to the RNAseq transcriptomic data. Diseases and function analyses that were based on differentially expressed target molecules in the dataset predicted that lipid metabolism would be enhanced whereas inflammation was predicted to be inhibited in SPI-fed compared to CAS-fed rats at 16 weeks. Combining upstream regulator and regulator effects functions in IPA facilitates the prediction of upstream regulators (e.g., transcription regulators) that could play important roles in attenuating or promoting liver steatosis due to SPI or CAS diets. Upstream regulators that were predicted to be activated (from expression of down-stream targets) linked to increased conversion of lipid and transport of lipid in SPI-fed rats included hepatocyte nuclear factor 4 alpha (HNF4A) and aryl hydrocarbon receptor (AHR). Upstream regulators that were predicted to be activated in CAS-fed rats linked to activation of phagocytosis and neutrophil chemotaxis included colony stimulating factor 2 and tumor necrosis factor. The results provide clear indication that long-term SPI-fed rats exhibited diminished inflammatory response and increased lipid transport in liver compared to CAS-fed rats that likely would contribute to reduced liver steatosis in this obese Zucker rat model.

scholarly journals Effects of Obesity and Short-Term Metformin Treatment on Liver Steatosis in Female Zucker Rats

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1640-1640
Author(s):  
Reza Hakkak ◽  
David Irby ◽  
Shannon Rose ◽  
Sirish Bennuri ◽  
Beverly Spray ◽  
...  
Keyword(s):  
Liver Disease ◽  
Rat Model ◽  
Liver Steatosis ◽  
Serum Markers ◽  
Zucker Rats ◽  
Metformin Treatment ◽  
Zucker Rat ◽  
Short Term ◽  
Obese Zucker Rat ◽  
Obese Rats

Abstract Objectives Non-alcoholic fatty liver disease (NAFLD) is the leading cause of liver disease in adolescents in the US and World, and the risk has increased with the rise in obesity. We reported that obesity increases Fatty liver (steatosis) using an obese Zucker rat model. Metformin is an oral anti-hyperglycemic agent approved by the FDA to treat type 2 diabetes (T2D) in adults and children older than 10 years of age. There is insufficient evidence regarding the effects of metformin in pediatric liver steatosis. The objective of this study was to investigate the effects of short-term metformin treatment on liver steatosis and related serum markers for liver damage. Methods Five week old lean (n = 16) and obese (n = 16) female Zucker rats after one week of acclimation, received AIN-93 G diet for 8 weeks to induce NAFLD. After 8 weeks, lean and obese rats were randomly assigned to the following four groups (8 rats/group): 1) lean without metformin (LC), 2) lean with metformin (LMet), 3) obese without metformin (ObC), and 4) obese with metformin (ObMet). Metformin were mixed with AIN-93 G diet at 1000 mg/kg of diet. Rats were weighed twice per week. All rats were sacrificed 10 weeks post-metformin treatment and serum and livers were collected. Steatosis was semiquantitated as a score of 1 to 4 based upon the relative degree of steatosis within hepatocytes: 1) &lt; 25%, 2) 25–50%, 3) 50–75%, and 4) &gt;75%. Serums were collected to measure the levels of Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT) on a clinical analyzer. Results Obese rats gained significantly more weight (P &lt; .001) than lean rats for both control and Metformin treatment groups and there was no significant difference between ObC vs. ObMet group (P = 0.20). The mean + SD liver steatosis scores for the LC, LMet, ObC and ObMet groups were 0.13 + o.3, 0.13 + o.3, 3.67 + 0.52 and 3.00 + 0.82. The ObMet treated rats had lower (P &lt; 0.04) liver steatosis than ObC rats. There were no significant differences for the serum ALT and AST levels between groups. However, obesity increased significantly (P &lt; 0.01) serum AST levels compared to LC but not in the metformin group. Conclusions In summary, in the obese zucker rat model, short-term metformin treatment decreased liver steatosis but did not impact serum markers of liver steatosis. Funding Sources Arkansas Children Research Institute/Arkansas Bioscience Institute.

A diet containing a high- versus low-daidzein level does not protect against liver steatosis in the obese Zucker rat model

2017 ◽  
Vol 8 (3) ◽  
pp. 1293-1298 ◽  
Author(s):  
Andrea Bell ◽  
Soheila Korourian ◽  
Huawei Zeng ◽  
Joshua Phelps ◽  
Reza Hakkak
Keyword(s):  
Rat Model ◽  
Liver Steatosis ◽  
Zucker Rat ◽  
Obese Zucker Rat ◽  
Body Weights

Low daidzeinversushigh daidzein mean (±SD) body weights over 8 weeks.

scholarly journals Effects of Short-Term Metformin Treatment on Gut Microbiota Profile Using Female Obese Zucker Rat Model

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 1213-1213
Author(s):  
Reza Hakkak ◽  
Chris Randolph ◽  
Sirish Bennuri ◽  
Michael Robeson
Keyword(s):  
Gut Microbiota ◽  
Rat Model ◽  
Zucker Rats ◽  
Metformin Treatment ◽  
Zucker Rat ◽  
Short Term ◽  
Fecal Samples ◽  
Obese Zucker Rat ◽  
Significant Difference ◽  
Obese Control

Abstract Objectives The correlation of short-term metformin treatment and specific alterations to the gut microbiota in obese models is less known. So, the objectives of this experiment was to investigate the effects of short-term metformin treatment on population of gut microbiota profile in obese rat model. Methods Five week old obese (n = 16) female Zucker rats after one week of acclimation, received AIN-93 G diet for 8 weeks and then rats were randomly assigned 8 rats/group): to 1) obese without metformin (ObC), or 2) obese with metformin (ObMet). Metformin were mixed with AIN-93G diet at 1000 mg/kg of diet. Rats were weighed twice per week. All rats were sacrificed 10 weeks post-metformin treatment and fecal samples were collected and kept at − 80c. Total microbial DNA were collected directly from the fecal samples using a PowerSoil® DNA isolation kit. Isolated DNA were used for shotgun-metagenomics data collection using Illumina NextSeq500 and analyzed using MetaPlAn and HUMAnN. DEICODE and Songbird used calculate log-ratios and differential ranks of taxa and functional pathways associated with metformin treatment respectively. The were then visualized using Qurro. Results There was no significant difference between ObC vs. ObMet group body weight (P = 0.20). Overall microbial beta-diversity (DEICODE), showed significant separation between the obese control and metformin samples (P = 0.0007). Differential ranking (Songbird) of Bacteroides dorei and B. massiliensis vs. all other Bacteroides spp., revealed that B. dorei and B. massiliensis were enriched in the obese metformin group, while the remaining Bacteroides spp. where enriched in the obese control group (P = 0.002). The differential ranking of pathway diversity contributed by the Bacteroides were also associated with treatment group (P = 0.008). Conclusions In summary, in the obese zucker rat model, short-term metformin treatment changes the gut microbiota profile. Funding Sources Arkansas Biosciences Institute.

scholarly journals Effects of Short and Long-Term Soy Protein Isolate Intake on Hepatic Cytochrome P450 Expression in Obese Zucker Rats

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 1225-1225
Author(s):  
Melisa Kozaczek ◽  
Walter Bottje ◽  
Reza Hakkak
Keyword(s):  
Gene Expression ◽  
Soy Protein ◽  
Soy Protein Isolate ◽  
Global Gene Expression ◽  
Protein Isolate ◽  
Zucker Rats ◽  
Z Score ◽  
Short And Long Term ◽  
Obese Zucker Rats

Abstract Objectives To determine the effects feeding for 8 (short-term) and 16 weeks (long-term) soy protein isolate on hepatic CYP gene expression. Methods 7-weeks old rats were randomly assigned to either a casein (CAS) or a soy protein isolate (SPI) diet. They were provided the diets ad libitum for 8 and 16 weeks. Rats were euthanized and livers were stored at − 80°C. RNA was extracted from liver samples, and sequenced to obtain transcriptomic data (RNAseq). Ingenuity Pathway Analysis software (IPA, Qiagen, CA) was used in the analysis of global gene expression data. This analysis includes predictions of activation or inhibition of molecules or upstream regulators and functions based on a generated z-score and p-value of overlap (P = 0.05). Z-scores were consider significant when &gt; 2 (activation) and &lt; −2 (inhibition). Results Comparing short- vs long-term feeding revealed an increase in the number of down-regulated CYP genes from only 3 at 8 weeks of SPI diet to 10 at 16 weeks of same diet (P &lt; 0.05). In contrast, upregulated CYP gene numbers showed a small increase in long-term SPI diet compared to short-term, from 14 genes at 8 weeks to 17 genes at 16 weeks (P &lt; 0.05). In addition, we present a predicted activation of the transcription factor Aryl hydrocarbon receptor (AHR, activation z-score = 2.146, P = 4.20E-11), linked to the subsequent activation or up-regulation of various CYPs genes, indirectly leading to the activation and inhibition of two main metabolic functions under SPI feeding: conversion of lipid (lipid metabolism) –predicted to be activated (z-score = 2.089, P = 2.77E-08), and recruitment of phagocytes (inflammatory response) –predicted to be inhibited (z-score = −2.311, P = 2.10E-05). Conclusions Through global gene expression analysis we showed that gene expression of drug-metabolizing cytochrome P450 genes was modified in genetically obese Zucker rats after being fed a soy-based diet for short- and long-term, and that this change could have an important role in attenuation of liver steatosis. Further research is needed to corroborate these results. Funding Sources This study was supported in part by the College of Medicine's University Medical Group (RH) and the Arkansas Biosciences Institute (WB, RH).

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