Shokri-Afra Hajar, Yousefi Abdolmaleki Elham, Mousavi Sadr Jadidi Elnaz Sadat, Oladi Ziaeddin, Moradi-Sardareh Hemen, Nabi Afjadi Mohsen, Ilbeigi Davod, Barmaki Haleh
Gut and Liver Research Center, Non-Communicable Diseases Institute, Mazandaran University of Medical Sciences, Sari, Iran.
Department of Internal Medicine, School of Medicine Ghaem Shahr, Razi Hospital, Ghaemshahr, Iran.
Mol Biol Rep. 2025 May 25;52(1):503. doi: 10.1007/s11033-025-10587-z.
The prevalence of nonalcoholic fatty liver disease (NAFLD) is rising at an alarming rate, making it a major global public health problem. The main pathophysiology of NAFLD is elevated de novo lipogenesis (DNL) in hepatocytes which leads to lipid accumulation. Because of their function in controlling DNL, sirtuin 1 (SIRT1) and AMP-activated protein kinase (AMPK) have been considered viable therapy targets for reduce lipid accumulation.
We examined the impact of the citrus flavonoid nobiletin (NOB) on the SIRT1-AMPK signaling pathway. This study involved incubating HepG2 cells with varying concentrations of NOB, measuring SIRT1 gene expression using qRT-PCR, assessing SIRT1 enzyme activity using a fluorometric assay, determining SIRT1 protein and AMPK phosphorylation levels by Western blotting, and measuring the lipid profile using semi- and quantitative assays. The results demonstrated that NOB significantly induced SIRT1 mRNA, protein expression, and activity similar to resveratrol (RSV) (as positive controls); additionally, NOB increased the phosphorylation of AMPK. EX-527 (negative control) significantly reversed the stimulatory effect of NOB on SIRT1 and AMPK. On the other hand, NOB decreased total lipid accumulation in cells exposed to oleic acid (OA) and reduced TG content to a normal level. However, the observed results on lipid profile were counteracted in the presence of EX-527.
NOB might be a new therapeutic approach for lipid accumulation management due to inducing the SIRT1-AMPK signaling pathway, however, it requires further investigations.
非酒精性脂肪性肝病（NAFLD）的患病率正以惊人的速度上升，使其成为一个主要的全球公共卫生问题。NAFLD的主要病理生理学是肝细胞中从头脂肪生成（DNL）增加，这导致脂质积累。由于其在控制DNL中的作用，沉默调节蛋白1（SIRT1）和AMP激活的蛋白激酶（AMPK）被认为是减少脂质积累的可行治疗靶点。
我们研究了柑橘类黄酮川陈皮素（NOB）对SIRT1-AMPK信号通路的影响。本研究包括用不同浓度的NOB孵育HepG2细胞，使用qRT-PCR测量SIRT1基因表达，使用荧光测定法评估SIRT1酶活性，通过蛋白质印迹法测定SIRT1蛋白和AMPK磷酸化水平，以及使用半定量和定量测定法测量脂质谱。结果表明，NOB显著诱导SIRT1 mRNA、蛋白表达和活性，类似于白藜芦醇（RSV）（作为阳性对照）；此外，NOB增加了AMPK的磷酸化。EX-527（阴性对照）显著逆转了NOB对SIRT1和AMPK的刺激作用。另一方面，NOB降低了暴露于油酸（OA）的细胞中的总脂质积累，并将甘油三酯含量降低到正常水平。然而，在存在EX-527的情况下，观察到的脂质谱结果被抵消。
由于诱导SIRT1-AMPK信号通路，NOB可能是一种管理脂质积累的新治疗方法，然而，这需要进一步研究。
