N-acetylcysteine influence on PI3K/Akt/mTOR and sphingolipid pathways in rats with MASLD induced by HFD: a promising new therapeutic purpose

IF 3.8 3区 医学 Q2 CELL BIOLOGY
Klaudia Sztolsztener, Daniel Michalak, Adrian Chabowski
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Abstract

Sphingolipid and glucose metabolism play important roles in the induction and progression of severe liver disorders like metabolic dysfunction-associated steatotic liver disease (MASLD). The perturbation in sphingolipid formation may improve the liver structure and functioning and may constitute the potential therapeutic options for the development of simple steatosis and its progression to steatohepatitis. This study aims to assess the influence of N-acetylcysteine (NAC) on the sphingolipid and insulin signaling pathways in rats subjected to standard or high-fat diets. Sphingolipid level was measured using high-performance liquid chromatography (HPLC). A multiplex assay kit determined the level of phosphorylated form of proteins included in the PI3K/Akt/mTOR pathway. The immunoblotting estimated the expression of proteins from sphingolipid and insulin transduction pathways. A histological Oil red O staining was used to assess the hepatic accumulation of lipid droplets. Molecular docking was applied to showcase NAC interaction with PI3K/Akt/mTOR pathway proteins. NAC decreased dihydroceramide and ceramide levels and increased phosphorylation of sphingosine and sphinganine. This antioxidant also enhanced phosphorylated Akt, GSK3α/β, and P70 S6 kinase and decreased phosphorylated S6RP. In silico docking analysis of insulin signaling molecules evidenced the higher binding affinity of NAC with all tested proteins, i.e., IRS1, PTEN, Akt, GSK3α/β, P70 S6 kinase, and S6RP, suggesting a potential protective influence on insulin resistance development, which is one of the criteria for MASLD diagnosing. Based on these data, NAC improved the hepatic insulin sensitivity and sphingolipid synthesis and storage, improving and restoring glucose homeostasis.

Abstract Image

N-乙酰半胱氨酸对高氟酸膳食诱导的 MASLD 大鼠 PI3K/Akt/mTOR 和鞘脂通路的影响:一种前景广阔的新治疗方法
鞘脂和葡萄糖代谢在代谢功能障碍相关性脂肪性肝病(MASLD)等严重肝病的诱发和进展中发挥着重要作用。干扰鞘脂的形成可改善肝脏的结构和功能,并可能成为治疗单纯性脂肪变性及其进展为脂肪性肝炎的潜在方案。本研究旨在评估 N-乙酰半胱氨酸(NAC)对标准或高脂饮食大鼠鞘磷脂和胰岛素信号通路的影响。采用高效液相色谱法(HPLC)测量鞘脂水平。多重检测试剂盒测定了 PI3K/Akt/mTOR 通路中蛋白质的磷酸化水平。免疫印迹测定了鞘脂和胰岛素转导通路蛋白的表达。组织学红油染色用于评估肝脏脂滴的积累。分子对接法显示了 NAC 与 PI3K/Akt/mTOR 通路蛋白的相互作用。NAC 降低了二氢甘油酰胺和神经酰胺的水平,增加了鞘磷脂和鞘氨醇的磷酸化。这种抗氧化剂还能增强磷酸化的 Akt、GSK3α/β 和 P70 S6 激酶,降低磷酸化的 S6RP。胰岛素信号分子的硅对接分析表明,NAC与所有测试蛋白(即IRS1、PTEN、Akt、GSK3α/β、P70 S6激酶和S6RP)的结合亲和力更高,这表明NAC对胰岛素抵抗的发展具有潜在的保护作用,而胰岛素抵抗是MASLD的诊断标准之一。基于这些数据,NAC改善了肝脏胰岛素敏感性和鞘脂的合成与储存,改善并恢复了糖稳态。
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来源期刊
Molecular and Cellular Endocrinology
Molecular and Cellular Endocrinology 医学-内分泌学与代谢
CiteScore
9.00
自引率
2.40%
发文量
174
审稿时长
42 days
期刊介绍: Molecular and Cellular Endocrinology was established in 1974 to meet the demand for integrated publication on all aspects related to the genetic and biochemical effects, synthesis and secretions of extracellular signals (hormones, neurotransmitters, etc.) and to the understanding of cellular regulatory mechanisms involved in hormonal control.
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