Biochanin A ameliorated oleate-induced steatosis in HepG2 cells by activating the SIRT3/AMPK/ULK-1 signaling pathway
Biochanin A ameliorated oleate-induced steatosis in HepG2 cells by activating the SIRT3/AMPK/ULK-1 signaling pathway
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Biochanin A 通过激活 SIRT3/AMPK/ULK-1 信号通路改善 HepG2 细胞中油酸诱导的脂肪变性
DOI：
10.1111/jfbc.14428
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发表时间：
2022
影响因子：
4
通讯作者：
Yan-Fen Chen
中科院分区：
文献类型：
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作者：
Guo-En Wang;Xiao-Ting Liu;Fan Yang;Ruo-Hong Wang;Xin-Yu Liu;Xi-Ting Lv;Xiao-Li Lin;Yan-Fen Chen
鹰嘴豆素A（Bio-A）是鹰嘴豆中富含的一种维生素，具有降血糖、降血脂和抗炎作用。然而，Bio‐A是否具有抗脂肪肝作用仍不清楚。本研究旨在评价Bio‐A对油酸（OA）处理的肝细胞的抗肝脂肪变性作用，并探讨其潜在机制。当与OA孵育24小时时，用不同浓度的Bio‐A处理HepG 2细胞24小时，以获得最佳抗脂肪肝剂量。用AMP活化蛋白激酶（AMPK）抑制剂化合物C或沉默调节蛋白-3（SIRT 3）抑制剂3-TYP处理HepG 2细胞，并用50 μM Bio‐A孵育。结果表明，在Bio‐A处理的脂肪肝细胞中，12.6%的脂质含量，特别是11.0%的甘油三酯含量，以及脂肪细胞分化相关蛋白的表达显著降低，随后Beclin 1的表达，Unc‐51‐样激酶1（ULK‐1）的磷酸化，微管相关蛋白1轻链3（LC 3）‐II/LC 3 ‐I比值，和p62表达的降低。结果表明，Bio‐A上调自噬体形成和自噬通量。此外，Bio‐A增加了OA处理的HepG 2细胞中的SIRT 3表达和AMPK磷酸化。AMPK和SIRT 3的阻断阻断了Bio‐A的抗脂肪肝作用和ULK‐1活化。AMPK抑制没有消除Bio‐A对SIRT 3的激活。AutoDock分析表明Bio‐A和SIRT 3之间可能存在相互作用。总之，Bio‐A通过激活SIRT 3/AMPK/ULK‐1介导的自噬减少OA处理的HepG 2细胞中的脂肪积累。这一发现为Bio-A对肝脂肪变性相关疾病的作用提供了理论依据。实际应用本研究突出了鹰嘴豆芽素A（Bio-A）对油酸（OA）处理的肝细胞的抗肝脂肪变性作用。Bio-A，鹰嘴豆属中的一种，具有减肥、抗炎、降血糖、降血脂等多种生物活性。本研究提供了Bio‐A治疗肝脂肪变性的新应用，并揭示了Bio‐A参与SIRT 3/AMPK/ULK‐1‐介导的自噬激活的潜在机制。这一发现为Bio-A在肝脏脂肪变性相关疾病中的应用提供了理论依据。
Biochanin A (Bio‐A), an isoflavone abundant in chickpeas, possesses hypoglycemic, hypolipidemic, and anti‐inflammatory effects. However, whether Bio‐A has antihepatosteatosis effect remains unclear. This study aimed to evaluate the antihepatosteatosis effect of Bio‐A on oleate (OA)‐treated hepatocytes, and explore the underlying mechanism. When incubated with OA for 24 h, HepG2 cells were treated with various concentrations of Bio‐A for 24 h to obtain an optimal antihepatosteatosis dose. HepG2 cells were treated with the AMP‐activated protein kinase (AMPK) inhibitor Compound C, or the sirtuin‐3 (SIRT3) inhibitor 3‐TYP, and incubated with 50 μM Bio‐A. The results indicated that 12.6% of lipid content, particularly 11.0% of triglyceride content, and the expression of adipocyte differentiation‐related protein were significantly decreased in Bio‐A‐treated hepatosteatosis cells, followed by an increase in the expression of Beclin 1, phosphorylation of Unc‐51‐like kinase 1 (ULK‐1), the microtubule‐associated protein 1 light chain 3 (LC3)‐II/LC3‐I ratio, and a decrease in expression of p62. The results indicated that Bio‐A upregulated autophagosome formation and autophagy flux. In addition, Bio‐A increased SIRT3 expression and AMPK phosphorylation in OA‐treated HepG2 cells. Blockade of AMPK and SIRT3 blocked the antihepatosteatosis effect and ULK‐1 activation by Bio‐A. AMPK inhibition did not eliminate the activation of SIRT3 by Bio‐A. AutoDock analysis demonstrated that interaction might exist between Bio‐A and SIRT3. In conclusion, Bio‐A reduced fat accumulation in OA‐treated HepG2 cells by activating SIRT3/AMPK/ULK‐1‐mediated autophagy. The findings provide a theoretical basis for the effect of Bio‐A on hepatic steatosis‐related diseases.Practical applicationsThis study highlights the antihepatosteatosis effects of biochanin A (Bio‐A) on oleate (OA)‐treated hepatocytes. Bio‐A, one of the isoflavones inCicer arietinumLinn., possesses multiple bioactivities such as antiobesity, anti‐inflammation, and hypoglycemic and hypolipidemic effects. This study provides a new application of Bio‐A to treat hepatic steatosis, and revealed the underlying mechanism of Bio‐A involved in the activation of the SIRT3/AMPK/ULK‐1‐mediated autophagy. The findings provide a theoretical basis for the application of Bio‐A to hepatic steatosis‐related diseases.