遗传 ›› 2026, Vol. 48 ›› Issue (1): 102-115.doi: 10.16288/j.yczz.25-173

• 研究报告 • 上一篇    

GPX8抑制猪骨骼肌卫星细胞成肌分化并促进慢肌纤维形成

郑腾飞(), 梁歆悦, 孟迎迎, 安亚龙, 王育禾, 史新娥, 李晓()   

  1. 西北农林科技大学动物科技学院,陕西省动物遗传育种与繁殖重点实验室,杨凌 712000
  • 收稿日期:2025-06-11 修回日期:2025-09-18 出版日期:2026-01-20 发布日期:2025-10-14
  • 通讯作者: 李晓,博士,副教授,研究方向:猪遗传育种研究。E-mail: nicelixiao@nwafu.edu.cn
  • 作者简介:郑腾飞,硕士研究生,专业方向:动物遗传育种与繁殖。E-mail: ztf455224189@outlook.com
  • 基金资助:
    农业生物育种国家科技重大专项(2023ZD0404702);陕西省创新能力支撑计划项目(2023-CX-TD-57);国家生猪产业技术体系(CARS-35-PIG)

GPX8 inhibits myogenic differentiation and promotes slow myofiber formation of porcine skeletal muscle satellite cells

Tengfei Zheng(), Xinyue Liang, Yingying Meng, Yalong An, Yuhe Wang, Xin’e Shi, Xiao Li()   

  1. Shaanxi Provincial Key Laboratory of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northwest A&F University, Yangling 712000, China
  • Received:2025-06-11 Revised:2025-09-18 Published:2026-01-20 Online:2025-10-14
  • Supported by:
    Biological Breeding-National Science and Technology Major Project(2023ZD0404702);Program for Shaanxi Science & Technology(2023-CX-TD-57);Earmarked Fund for(CARS-35-PIG)

摘要:

谷胱甘肽过氧化物酶8 (glutathione peroxidase 8,GPX8)是谷胱甘肽过氧化物酶家族的重要成员。全基因组关联分析(genome-wide association analysis,GWAS)表明,GPX8与猪生长发育及胴体性状高度相关。本研究以猪骨骼肌卫星细胞为研究对象,分别干扰和过表达GPX8,利用免疫荧光染色、qRT-PCR和Western blotting等方法探究GPX8对成肌分化和肌纤维类型转化的影响。结果显示,干扰GPX8显著提高成肌分化指数(P<0.01),促进分化标志基因MyHCMyoG mRNA和蛋白的表达(P<0.05),过表达GPX8得到相反的结果。干扰GPX8显著降低MYH7 mRNA(P<0.01)和慢肌纤维蛋白(slow-twitch MyHC,slow-MyHC)的表达(P<0.05),抑制线粒体生物发生,过表达GPX8得到相反的结果。整合GWAS等多组学数据筛选出调控GPX8表达的顺式eQTLs (expression quantitative trait locus),并通过双荧光素酶报告分析检测候选SNPs对GPX8启动子活性的影响,鉴定出GPX8启动子区5个候选SNPs (rs335618489、rs325233940、rs32989756、rs322106839和rs701033890),其中rs335618489-T、rs325233940-G、rs32989756-T和rs322106839-G通过改变启动子活性显著上调GPX8表达水平(P<0.01),进而影响猪产肉性状。总之,本研究表明GPX8抑制猪骨骼肌卫星细胞成肌分化并促进快肌纤维向慢肌纤维类型转化,GPX8启动子区功能性SNPs通过调控GPX8表达影响猪肌肉发育,为进一步提高猪肉产量提供了育种靶点。

关键词: 猪骨骼肌卫星细胞, GPX8, 成肌分化, 肌纤维类型转化

Abstract:

Glutathione peroxidase 8(GPX8) is a key member of the glutathione peroxidase family. Genome-wide association studies (GWAS) have indicated that GPX8 is highly associated with growth and carcass traits of pigs. Using porcine skeletal muscle satellite cells, this study explored the effects of GPX8 on myogenic differentiation and myofiber type switching through GPX8 knockdown and overexpression, combined with analyses involving immunofluorescence staining, qRT-PCR, and Western blotting. The results demonstrated that GPX8 knockdown significantly increased the myogenic differentiation index (P<0.01) and promoted both mRNA and protein levels of the myogenic marker genes MyHC and MyoG (P<0.05). Conversely, GPX8 overexpression exhibited the opposite effects. GPX8 knockdown significantly reduced the mRNA level of MYH7 (P<0.01) and protein level of slow-twitch MyHC (slow-MyHC)(P<0.05), while suppressing mitochondrial biogenesis. In contrast, GPX8 overexpression exhibited opposing results. Integrated multi-omics data from GWAS analyses were employed to identify expression quantitative trait locus (eQTLs) regulating GPX8 expression. The effects of candidate SNPs on GPX8 promoter activity were further validated using dual-luciferase reporter assays. Five candidate SNPs (rs335618489, rs325233940, rs32989756, rs322106839, and rs701033890) were identified within the GPX8 promoter region. Among these, rs335618489-T, rs325233940-G, rs32989756-T, and rs322106839-G significantly upregulated GPX8 expression level by altering promoter activity (P<0.01), thereby influencing porcine muscle development traits. In summary, this study demonstrates that GPX8 inhibits the myogenic differentiation of porcine skeletal muscle satellite cells and promotes the transition from fast-twitch to slow-twitch myofibers. Functional SNPs in the GPX8 promoter region influence porcine muscle development by modulating GPX8 expression, thereby providing valuable breeding targets for improving pork production.

Key words: porcine skeletal muscle satellite cells, GPX8, myogenic differentiation, myofiber type transformation