Graduate Program of Health Sciences (PPGCS), Federal University of Sergipe, Aracaju, Sergipe, Brazil.
Laboratory of Nutritional Genomics, School of Applied Sciences, University of Campinas, Pedro Zaccaria, 1300 Zip, Limeira, 13484-350, Brazil.
Trials. 2021 Dec 18;22(1):927. doi: 10.1186/s13063-021-05702-x.
The low-grade inflammation is pivotal in obesity and its comorbidities; however, the inflammatory proteins are out of target for traditional drug therapy. Omega-3 (ω3) fatty acids can modulate the downstream signaling of Toll-like receptor (TLR) and tumor necrosis factor-α receptor (TNFα) through GPR120, a G-protein-coupled receptor, a mechanism not yet elucidated in humans. This work aims to investigate if the ω3 supplementation, at a feasible level below the previously recommended level in the literature, is enough to disrupt the inflammation and endoplasmic reticulum stress (ER-stress), and also if in acute treatment (3 h) ω3 can activate the GPR120 in peripheral blood mononuclear cells (PBMC) and leukocytes from overweight non-alcoholic fatty liver disease (NAFLD) participants. The R270H variant of the Ffar4 (GPR120 gene) will also be explored about molecular responses and blood lipid profiles. A triple-blind, prospective clinical trial will be conducted in overweight men and women, aged 19-75 years, randomized into placebo or supplemented (2.2 g of ω3 [EPA+DHA]) groups for 28 days. For sample calculation, it was considered the variation of TNFα protein and a 40% dropout rate, obtaining 22 individuals in each group. Volunteers will be recruited among patients with NAFLD diagnosis. Anthropometric parameters, food intake, physical activity, total serum lipids, complete fatty acid blood profile, and glycemia will be evaluated pre- and post-supplementation. In the PBMC and neutrophils, the protein content and gene expression of markers related to inflammation (TNFα, MCP1, IL1β, IL6, IL10, JNK, and TAK1), ER-stress (ATF1, ATF6, IRE1, XBP1, CHOP, eIF2α, eIF4, HSP), and ω3 pathway (GPR120, β-arrestin2, Tab1/2, and TAK1) will be evaluated using Western blot and RT-qPCR. Participants will be genotyped for the R270H (rs116454156) variant using the TaqMan assay. It is hypothesized that attenuation of inflammation and ER-stress signaling pathways in overweight and NAFLD participants will be achieved through ω3 supplementation through binding to the GPR120 receptor. TRIAL REGISTRATION: ClinicalTrials.gov #RBR-7x8tbx. Registered on May 10, 2018, with the Brazilian Registry of Clinical Trials.
低度炎症在肥胖及其合并症中起着关键作用；然而，炎症蛋白是传统药物治疗的靶点之外。ω-3（ω3）脂肪酸可以通过 Toll 样受体（TLR）和肿瘤坏死因子-α 受体（TNFα）的下游信号转导来调节 G 蛋白偶联受体（GPCR）GPR120，这一机制在人类中尚未阐明。本研究旨在探讨 ω3 补充剂是否能在低于文献中先前推荐的水平下达到足够的效果，以破坏炎症和内质网应激（ER 应激），并且如果在急性治疗（3 小时）中，ω3 能激活超重非酒精性脂肪性肝病（NAFLD）患者外周血单核细胞（PBMC）和白细胞中的 GPR120。还将探讨 Ffar4（GPR120 基因）的 R270H 变体在分子反应和血液脂质谱方面的作用。一项针对超重男性和女性的、随机、双盲、前瞻性临床试验将进行，年龄在 19-75 岁之间，分为安慰剂或补充（2.2 克 ω3[EPA+DHA]）组，为期 28 天。为了进行样本计算，考虑了 TNFα 蛋白的变化和 40%的失访率，每组获得 22 名参与者。志愿者将从 NAFLD 诊断患者中招募。在补充前后，将评估人体测量参数、饮食摄入、体力活动、总血清脂质、全血脂肪酸谱、血糖。在 PBMC 和中性粒细胞中，通过 Western blot 和 RT-qPCR 评估与炎症（TNFα、MCP1、IL1β、IL6、IL10、JNK 和 TAK1）、ER 应激（ATF1、ATF6、IRE1、XBP1、CHOP、eIF2α、eIF4、HSP）和 ω3 途径（GPR120、β-arrestin2、Tab1/2 和 TAK1）相关的标志物的蛋白含量和基因表达。使用 TaqMan 检测法对 R270H（rs116454156）变体进行基因型分析。假设通过与 GPR120 受体结合，ω3 补充剂将通过结合 GPR120 受体，减轻超重和 NAFLD 参与者的炎症和 ER 应激信号通路。试验注册：ClinicalTrials.gov #RBR-7x8tbx。于 2018 年 5 月 10 日在巴西临床试验注册处注册。
