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肥胖性肝脂肪变性大鼠模型中,连续 8 周给予大豆蛋白分离物或酪蛋白为基础的饮食后,采用 RNAseq 和 PCR 分析比较肝脏基因表达。

Comparison of liver gene expression by RNAseq and PCR analysis after 8 weeks of feeding soy protein isolate- or casein-based diets in an obese liver steatosis rat model.

机构信息

Department of Poultry Science & The Center of Excellence for Poultry Science, University of Arkansas, 1260 W. Maple, Fayetteville, AR 72701, USA.

出版信息

Food Funct. 2019 Dec 11;10(12):8218-8229. doi: 10.1039/c9fo01387c.

Abstract

Previously, we reported that feeding soy protein isolate (SPI) reduced liver steatosis in obese rats compared to those fed a casein (CAS)-based diet; however, the mechanism for this protection is unknown. To gain insight into the ability of SPI to ameliorate liver steatosis, we conducted transcriptomic (RNAseq) analysis on liver samples from obese rats fed either the SPI- or CAS-based diets (n = 8 per group) for 8 weeks using an Ilumina HiSeq with 100 base paired end reads for sequencing. Data were analyzed by Ingenuity Pathway Analysis (IPA) software using a P < 0.05 and 1.3-fold differential expression cutoff values between the SPI- and CAS-based groups. To independently validate the RNAseq data, we conducted targeted mRNA expression analysis using quantitative PCR (qPCR) on a subset of differently expressed genes. The results indicate that mRNA expression by qPCR concurred with RNAseq for NPTX2, GPT, INMT, and HAL that were up-regulated in SPI-fed rats (P < 0.05) and PRSS8, AJUBA, CSF2RB, and Cyp2c12 that were down-regulated (P < 0.05) in SPI-fed rats compared to CAS-fed rats. Our findings may shed light on understanding mechanisms enabling SPI diet to reduce liver steatosis in this obese Zucker rat model.

摘要

先前,我们报道了与基于酪蛋白(CAS)的饮食相比,给肥胖大鼠喂食大豆蛋白分离物(SPI)可减少肝脏脂肪变性;然而,这种保护作用的机制尚不清楚。为了深入了解 SPI 改善肝脏脂肪变性的能力,我们对喂食 SPI 或 CAS 饮食 8 周的肥胖大鼠的肝脏样本进行了转录组(RNAseq)分析(每组 n = 8),使用 Ilumina HiSeq 进行测序,读长为 100 个碱基对。使用 IPA 软件对数据进行了 Ingenuity Pathway Analysis(IPA)分析,使用 SPI 和 CAS 组之间的 P < 0.05 和 1.3 倍差异表达截断值进行分析。为了独立验证 RNAseq 数据,我们使用定量 PCR(qPCR)对一组差异表达基因进行了靶向 mRNA 表达分析。结果表明,qPCR 的 mRNA 表达与 RNAseq 一致,在 SPI 喂养的大鼠中上调的基因有 NPTX2、GPT、INMT 和 HAL(P < 0.05),在 SPI 喂养的大鼠中下调的基因有 PRSS8、AJUBA、CSF2RB 和 Cyp2c12(P < 0.05)。我们的研究结果可能有助于理解 SPI 饮食在肥胖 Zucker 大鼠模型中减少肝脏脂肪变性的机制。

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