摘要
[目的]本文旨在探究染料木黄酮(GEN)对棕榈酸(PA)诱导的鸡胚原代肝细胞脂代谢紊乱的调控作用及潜在机制。[方法]选用不同浓度GEN预处理PA诱导的鸡胚原代肝细胞,随后采用G蛋白偶联雌激素受体(GPR30)或AMP-依赖性蛋白激酶(AMPK)的抑制剂或拮抗剂等预处理鸡胚原代肝细胞,采用比色法检测细胞活力及甘油三酯(TG)和总胆固醇(TC)含量,采用实时荧光定量PCR和Western blot法检测脂代谢和线粒体功能相关因子的表达水平。[结果]与对照组相比,0.3 mmol·L^(-1) PA处理36 h以及0.4和0.5 mmol·L^(-1) PA处理24~48 h显著降低鸡胚原代肝细胞活力;GEN处理显著缓解PA诱导的鸡胚原代肝细胞TG和TC含量增加;GEN处理显著增加PA诱导的鸡胚原代肝细胞脂肪分解相关基因的表达水平,而明显降低脂肪合成相关基因的表达水平;GEN通过激活AMPK降低脂肪酸合成相关因子的活性,增加脂肪酸分解和线粒体氧化损伤相关因子的表达水平;GEN通过激活GPR30诱导AMPK信号的活化。[结论]GEN通过激活GPR30-AMPK信号通路调节脂肪代谢和线粒体氧化损伤途径中关键因子的表达,最终缓解PA诱导的鸡胚原代肝细胞脂代谢紊乱和线粒体氧化损伤。
[Objectives]This paper aimed to explore the regulatory effect and potential mechanism of genistein(GEN)on palmitic acid(PA)-induced lipid metabolism disorder in primary chicken hepatocytes.[Methods]Primary chicken hepatocytes induced by PA were pretreated with different concentrations of GEN.Next,primary chicken hepatocytes were pretreated with the inhibitor or antagonist of G protein-coupled estrogen receptor(GPR30)or AMP-dependent protein kinase(AMPK).Colorimetric assay was used to detect cell viability,triglyceride(TG)and total cholesterol(TC)contents.Real-time quantitative PCR and Western blot were used to determine the expression levels of the factors related to lipid metabolism and mitochondrial function.[Results]Compared with the control group,0.3 mmol·L^(-1) PA treatment for 36 h as well as 0.4 and 0.5 mmol·L^(-1) PA treatment for 24-48 h significantly decreased the cell viability of primary chicken hepatocytes.GEN treatment significantly alleviated the increase of TG and TC contents in primary chicken hepatocytes induced by PA.GEN treatment significantly increased the expression level of genes related to fat decomposition in primary chicken hepatocytes induced by PA,but markedly reduced the expression level of genes related to fat synthesis.GEN decreased the activity of factors related to fatty acid synthesis,and increased the expression level of factors related to fatty acid decomposition and mitochondrial oxidative damage by activating AMPK.GEN induced the activation of AMPK signal by activating GPR30.[Conclusions]GEN regulated the expression of key factors in lipid metabolism and mitochondrial oxidative damage pathway by activating the GPR30-AMPK signaling pathways,ultimately alleviating PA-induced lipid metabolism disorder and mitochondrial oxidative damage in primary chicken hepatocytes.
作者
贺复亚
王慧慧
公苹
赵淑娟
马海田
李龙龙
HE Fuya;WANG Huihui;GONG Ping;ZHAO Shujuan;MA Haitian;LI Longlong(College of Veterinary Medicine/Key Laboratory of Animal Physiology and Biochemistry,Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China)
出处
《南京农业大学学报》
北大核心
2025年第4期902-911,共10页
Journal of Nanjing Agricultural University
基金
国家自然科学基金项目(32202758)
中央高校基本科研业务费专项资金(KJYQ2025007)。
作者简介
通信作者:李龙龙,助理研究员,研究方向为物质代谢及信号转导机制,E-mail:lilonglong@njau.edu.cn。
