脂联素抑制AKT信号拮抗砷诱导肝细胞脂质沉积
diponectin inhibits AKT signaling and antagonizes arsenic-induced lipid deposition in hepatocytes
目的 探讨重组人脂联素（adiponectin, APN）对砷诱导肝HepG-2细胞脂质沉积的干预效应及其分子机制。方法 实验分为对照组、亚砷酸钠（SA）染毒组、rAPN干预+SA染毒组、蛋白激酶B（AKT）抑制剂（GSK690693）干预+SA染毒组，分别对体外培养的HepG-2细胞进行处理，通过细胞涂片油红O染色观察细胞脂质沉积特征，应用生物化学及酶联免疫吸附试验（ELISA）检测线粒体膜电位（ΔΨm）、线粒体肉碱棕榈酰转移酶 1(CPT-1)活性、游离脂肪酸（FFAs）和APN含量，免疫印迹检测磷酸化AKT（p-AKT）、线粒体谷胱甘肽S转移酶K1（GSTK1）和炎症因子半胱天冬酶1（Caspase-1）水平。 结果 相比对照组，SA染毒组显示细胞内脂质沉积，ΔΨm、CPT-1活性、GSTK1和APN水平下降，而FFAs、p-AKT和Caspase-1水平上调。相比SA染毒组，重组人APN干预后显示细胞脂质沉积程度减轻，ΔΨm、CPT-1活性、GSTK1和APN水平升高，而FFAs、p-AKT和Caspase-1水平下调。类似地，GSK690693干预后亦显示细胞脂质沉积程度减轻，ΔΨm、CPT-1活性、GSTK1水平升高，p-AKT和Caspase-1水平下调。 结论 砷诱导HepG-2细胞脂质沉积与AKT信号激活有关，脂联素可通过抑制AKT信号拮抗砷诱导肝细胞脂质代谢障碍及脂质沉积发生。<br />
Objective To investigate the effects and molecular mechanism of recombinant human adiponectin (APN) on arsenic induced lipid deposition in hepatic HepG-2 cells. Methods The experiments were divided into control group, sodium arsenite (SA) exposure group, rAPN intervention + SA exposure group, and protein kinase B (AKT) inhibitor (GSK690693) intervention + SA exposure group. After cultured HepG-2 cells in vitro were treated respectively, the characteristics of cellular lipid deposition were observed by oil red O staining of cells smears, and the methods of biochemical or enzyme-linked immunosorbent assay (ELISA) were applied to determine the levels of intracellular mitochondrial membrane potential (ΔΨm), mitochondrial carnitine palmitoyltransferase-1 (CPT-1) activity, free fatty acids (FFAs) and APN. Western-blotting (WB) was applied to detect the levels of phosphorylated AKT (p-AKT), mitochondrial Glutathione S-transferase K1 (GSTK1) and inflammatory factor Caspase-1. Results Compared with control group, SA exposed group showed intracellular lipid deposition and the decreased levels of mitochondria ΔΨm, CPT1 activity, GSTK1 or APN, while the levels of FFAs, p-AKT and Caspase-1 increased. Compared with SA exposed group, recombinant human APN (rAPN) intervention led to the alleviated lipid deposition in hepatocytes and the increased levels of ΔΨm, CPT1 activity, GSTK1 or APN, while the levels of FFAs, p-AKT and Caspase-1 decreased. Similarly, the intervention of AKT inhibitor showed also the reduced cellular lipid deposition and the upregulated levels of ΔΨm, CPT1 or GSTK1, while the levels of p-AKT and caspase-1 downregulated. Conclusion Arsenic induced lipid deposition in HepG-2 cells is associated with AKT signaling, and APN can antagonize arsenic-induced lipid metabolism disorder and lipid deposition in hepatocytes via inhibiting AKT signaling.<br />
基础医学生物化学分子生物学
砷HepG-2细胞脂质沉积脂联素蛋白激酶B
rsenicHepG-2 cellslipid depositionadiponectinprotein kinase B
.脂联素抑制AKT信号拮抗砷诱导肝细胞脂质沉积[EB/OL].(2023-03-05)[2025-08-05].https://chinaxiv.org/abs/202303.00026.点此复制
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