College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, P. R. China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, P. R. China.
J Nutr Biochem. 2022 Jul;105:109001. doi: 10.1016/j.jnutbio.2022.109001. Epub 2022 Mar 26.
Selenium (Se) is a vital minor element for the organism. Se deficiency caused inflammation in kidney tissue and regulate the expression of selenoproteins and microRNAs (miRNAs). Pyroptosis involved in the inflammatory response, however, whether microRNA targets GPX4 to regulate Se-deficient kidney tissue pyroptosis is unclear. In this study, broilers were divided into two groups, Control group with 0.3mg/kg Se diet and Se-deficient group with 0.03mg/kg Se diet. The dual luciferase reporter assay system and quantitative real-time PCR (qRT-PCR) were used to screen the specificity of miR-1656 and its target protein in Se-deficient broilers. We tested the pyroptosis-related genes of Se-deficient broilers kidney and miR-1656-transfected primary broilers kidney by qRT-PCR, Western blot (WB) and immunofluorescence staining. Our research indicated that the GPX4 is one of the target genes of miR-1656, and Se deficiency leaded to the overexpression of miR-1656 and the increased expression of pyroptosis-related genes. The overexpression of miR-1656 can induce increased expression of pyroptosis-related genes including NLRP3, Caspase-1, IL-18, and IL-1β by inhibiting the release of GPX4. This study showed that miR-1656 could increase the release of ROS by targeting GPX4, activated the NLRP3 inflammasome, and release the inflammatory factors IL-1β and IL-18 to trigger pyroptosis in the kidney tissue of Se-deficient broilers. This finding may provide new research ideas for kidney injury and cell death due to Se deficiency.
硒（Se）是生物体必需的微量元素。硒缺乏会引起肾脏组织炎症，并调节硒蛋白和 microRNAs（miRNAs）的表达。细胞焦亡参与炎症反应，然而，miRNA 是否通过靶向 GPX4 来调节硒缺乏的肾脏组织细胞焦亡尚不清楚。本研究将肉鸡分为两组，对照组饲粮含 0.3mg/kg Se，硒缺乏组饲粮含 0.03mg/kg Se。采用双荧光素酶报告基因检测系统和实时定量 PCR（qRT-PCR）筛选硒缺乏肉鸡中 miR-1656 及其靶蛋白的特异性。通过 qRT-PCR、Western blot（WB）和免疫荧光染色检测硒缺乏肉鸡肾脏的细胞焦亡相关基因和转染 miR-1656 的原代肉鸡肾脏的细胞焦亡相关基因。研究表明，GPX4 是 miR-1656 的靶基因之一，硒缺乏导致 miR-1656 过表达和细胞焦亡相关基因表达增加。miR-1656 的过表达通过抑制 GPX4 的释放，可诱导细胞焦亡相关基因包括 NLRP3、Caspase-1、IL-18 和 IL-1β 的表达增加。本研究表明，miR-1656 可以通过靶向 GPX4 增加 ROS 的释放，激活 NLRP3 炎性小体，释放炎性因子 IL-1β 和 IL-18，从而引发硒缺乏肉鸡肾脏组织的细胞焦亡。这一发现可能为硒缺乏引起的肾脏损伤和细胞死亡提供新的研究思路。
