Department of Dietetics, Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences, Nowoursynowska 159c, 02-776, Warsaw, Poland.
Biochemistry Division, Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159, 02-787, Warsaw, Poland.
Eur J Nutr. 2019 Oct;58(7):2859-2873. doi: 10.1007/s00394-018-1838-3. Epub 2018 Oct 4.
Beta-glucans are biologically active polysaccharides having antioxidant, immunomodulatory, and antiinflammatory properties. This study investigated the transcriptomic profile in peripheral blood of rats with LPS-induced enteritis, which were fed a diet supplemented with high- (G1) and low- (G2) molecular-weight oat beta-glucans.
Two-color rat gene expression microarrays were applied and the analysis was performed using a common reference design to provide easy means of comparing samples from various experimental conditions against one another. Common reference sample was labeled with cyanine 3 (Cy3) and investigated samples from each experimental group: C-G0 (control group fed semi-synthetic diet), LPS-G0 (LPS-challenged group fed semi-synthetic diet), LPS-G1 (LPS-challenged group fed G1 beta-glucan enriched diet), and LPS-G2 (LPS-challenged group fed G2 beta-glucan enriched diet) were labeled with cyanine 5 (Cy5). Each microarray was performed in quadruplicate. Statistical analysis was performed using one-way ANOVA and Tukey's HSD post-hoc test (p < 0.05). A multiple testing correction was performed using Benjamini and Hochberg False Discovery Rate < 5%. A quantitative real-time RT-PCR was performed to verify the expression of chosen transcripts.
The microarray analyses revealed differentially expressed transcripts between: the LPS-G0 and the control groups: C-G0 (138 genes), the LPS-G1 and LPS-G0 groups (533 genes), and the LPS-G2 and LPS-G0 groups (97 genes). Several differentially expressed genes in the beta-glucan-supplemented groups encoded proteins belonging to TLR and NLR signaling pathways, as well as prostaglandin synthesis and regulation pathways. Both beta-glucans up-regulated the expression of Atg10, which belongs to the family of autophagy-related genes, suggesting a possible link between autophagy induction and beta-glucan supplementation.
The changes in gene expression observed in the peripheral blood indicate that oat beta-glucans exerted a protective effect in rats with an induced inflammatory state caused by LPS challenge. The greater number of differentially expressed genes was observed in group supplemented with G1 beta-glucan, pointing at the differences in the mode of action of high- and low-molecular-weight beta-glucans in the organism.
β-葡聚糖是具有抗氧化、免疫调节和抗炎特性的生物活性多糖。本研究调查了脂多糖诱导的肠炎大鼠外周血的转录组谱，这些大鼠喂食添加了高分子量（G1）和低分子量（G2）燕麦β-葡聚糖的饮食。
应用双色大鼠基因表达微阵列，采用通用参考设计进行分析，以便于比较不同实验条件下的样本。通用参考样本用 Cy3（青色）标记，每个实验组的研究样本：C-G0（喂食半合成饮食的对照组）、LPS-G0（喂食半合成饮食的 LPS 挑战组）、LPS-G1（喂食富含 G1β-葡聚糖的饮食的 LPS 挑战组）和 LPS-G2（喂食富含 G2β-葡聚糖的饮食的 LPS 挑战组）用 Cy5（青色）标记。每个微阵列重复进行四次。使用单向方差分析和 Tukey 的 HSD 事后检验（p<0.05）进行统计分析。使用 Benjamini 和 Hochberg 错误发现率（False Discovery Rate，FDR）<5%进行多重测试校正。进行实时定量 RT-PCR 以验证所选转录物的表达。
微阵列分析显示 LPS-G0 与 C-G0（138 个基因）、LPS-G1 与 LPS-G0（533 个基因）和 LPS-G2 与 LPS-G0（97 个基因）之间存在差异表达的转录物。补充β-葡聚糖的组中几个差异表达的基因编码属于 TLR 和 NLR 信号通路以及前列腺素合成和调节通路的蛋白质。两种β-葡聚糖均上调了自噬相关基因家族成员 Atg10 的表达，表明自噬诱导和β-葡聚糖补充之间可能存在联系。
外周血中观察到的基因表达变化表明，燕麦β-葡聚糖对 LPS 挑战引起的炎症状态大鼠发挥了保护作用。补充 G1β-葡聚糖的组观察到更多差异表达的基因，表明高分子量和低分子量β-葡聚糖在体内的作用模式存在差异。
