水生生物学报2025，Vol.49Issue(12)：40-48,9.DOI:10.3724/1000-3207.2025.2025.0175
大黄鱼FFAR4受体介导ω-3与ω-6脂肪酸对细胞的差异性影响机制
DIFFERENTIAL CELLULAR RESPONSES TO ω-3 AND ω-6 FATTY ACIDS MEDIATED BY FFAR4 RECEPTOR IN LARGE YELLOW CROAKER(LARIMICHTHYS CROCEA)
摘要
Abstract
Free fatty acid receptor 4(FFAR4),a G protein-coupled receptor(GPCR),plays a key role in sensing long-chain polyunsaturated fatty acids(LC-PUFAs)in mammals.However,its ligand selectivity and downstream signaling mechanisms in fish remain poorly understood.In this study,we investigated the ligand recognition and signaling cha-racteristics of FFAR4 from the large yellow croaker(Larimichthys crocea),with particular emphasis on ω-3 and ω-6 fatty acids.A stable HEK293T cell line expressing LcFFAR4 was established to systematically assess ligand-induced receptor internalization,intracellular calcium flux,and cyclic adenosine monophosphate(cAMP)signaling pathway.Subcellular localization analysis revealed predominant plasma membrane distribution of LcFFAR4.Ligand stimulation assays demonstrated that both eicosapentaenoic acid(EPA,ω-3)and linoleic acid(LA,ω-6)induced receptor internali-zation,with EPA showing significantly stronger effect.Calcium imaging revealed low concentrations EPA markedly increased intracellular calcium levels,indicating activation of the calcium signaling pathway,whereas LA elicited no significant change.Dual-luciferase reporter assays further confirmed both EPA and LA activated the LcFFAR4-media-ted cAMP signaling pathway,with EPA displaying a more potent effect.Functional analyses revealed that EPA more effectively mitigated lipopolysaccharide(LPS)-induced inflammatory responses via LcFFAR4.These results collec-tively demonstrate that LcFFAR4 exhibits distinct ligand selectivity and signaling bias.EPA,an ω-3 fatty acid,func-tions as a more potent agonist than LA,inducing stronger receptor internalization and more robust activation of both calcium and cAMP signaling pathways.This study provides the first evidence of teleost FFAR4 differential responses to ω-3 and ω-6 fatty acids from a GPCR signaling perspective,offering new insights into the fatty acid sensing mecha-nisms and their potential roles in nutrient sensing and metabolic regulation in fish.关键词
FFAR4/长链多不饱和脂肪酸/内化/钙信号/cAMP/配体偏好性/大黄鱼Key words
FFAR4/Long-chain polyunsaturated fatty acids/Receptor internalization/Calcium signaling/cAMP/Ligand selectivity/Larimichthys crocea分类
农业科技引用本文复制引用
吴彩霞,李庆飞,隋昌序,艾庆辉..大黄鱼FFAR4受体介导ω-3与ω-6脂肪酸对细胞的差异性影响机制[J].水生生物学报,2025,49(12):40-48,9.基金项目
国家自然科学基金(32102807) （32102807）
山东省自然科学基金(ZR2024MC004) （ZR2024MC004）
国家重点研发计划(2023YFD2400601) （2023YFD2400601）
国家海水鱼产业技术体系项目(CARS-47-G10)资助[Supported by the National Natural Science Foundation of China(32102807) （CARS-47-G10）
Shandong Provincial Natural Science Foundation(ZR2024MC004) （ZR2024MC004）
the National Key Research and Development Program of China(2023YFD2400601) （2023YFD2400601）
the China Agriculture Research System for Marine Fish Culture Industry(CARS-47-G10)] （CARS-47-G10）
