Volume 46, Issue 12 e14428
ORIGINAL ARTICLE

Biochanin A ameliorated oleate-induced steatosis in HepG2 cells by activating the SIRT3/AMPK/ULK-1 signaling pathway

Guo-En Wang

Corresponding Author

Guo-En Wang

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

Correspondence

Guo-En Wang, School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou 510006, China.

Email: [email protected]; [email protected]

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Xiao-Ting Liu

Xiao-Ting Liu

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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Fan Yang

Fan Yang

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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Ruo-Hong Wang

Ruo-Hong Wang

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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Xin-Yu Liu

Xin-Yu Liu

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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Xi-Ting Lv

Xi-Ting Lv

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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Xiao-Li Lin

Xiao-Li Lin

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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Yan-Fen Chen

Yan-Fen Chen

School of Traditional Chinese medicine, Guangdong Pharmaceutical University, Guangzhou, China

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First published: 20 September 2022
Citations: 6

Guo-En Wang and Xiao-Ting Liu contributed equally to the work. Guo-En Wang and Xiao-Ting Liu are co-first author.

Abstract

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 applications

This study highlights the antihepatosteatosis effects of biochanin A (Bio-A) on oleate (OA)-treated hepatocytes. Bio-A, one of the isoflavones in Cicer arietinum Linn., 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.

CONFLICT OF INTEREST

The authors have no conflicts of interest to declare that are relevant to the content of this article.

DATA AVAILABILITY STATEMENT

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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