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Introduction <p>The higher prevalence of non-alcoholic fatty liver disease (NAFLD) in men than women before menopause and the reduced difference post-menopause suggest that sex hormones may influence liver lipid accumulation. This study compared the effects of sex hormones on lipid droplet (LD) accumulation in palmitate/ oleate-treated HepG2 cells.</p> Methods <p>The MTT method was used to determine effective doses of palmitic and oleic acids in HepG2 cells, followed by a combined dose for inducing LD formation. Changes in LD content after treatment with various doses of β-estradiol and testosterone were evaluated qualitatively and semi-quantitatively using Oil Red O staining and light microscopy. The effects of these hormones on gene expression related to LD formation and lipogenesis, including PLIN2, ATGL, CGI-58, and CIDEB, were assessed using quantitative PCR.</p> Results <p>Treatment of HepG2 cells with palmitate and oleate increased LD accumulation and the expression of PLIN2 and CIDE while elevating ATGL expression without affecting CGI-58. With no significant difference, both β-estradiol and testosterone significantly reduced LD accumulation in steatotic HepG2 cells. Gene analysis indicated that both hormones decreased PLIN2 and increased CGI-58 expression. Testosterone did not affect CIDE, while β-estradiol reduced it at low doses. Combined treatment showed no significant changes in gene expression compared to individual hormone effects, but LD accumulation was synergistically reduced.</p> Conclusion <p>This study demonstrates that β-estradiol and testosterone significantly modulate LD content and the expression of key regulatory genes in HepG2 cells, with β-estradiol showing a somewhat dominant role in enhancing lipid turnover and mitigating lipid accumulation.</p>

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Comparative analysis of β-Estradiol and testosterone on lipid droplet accumulation, and regulatory protein expression in palmitate/oleate-induced fatty HepG2 cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年4月16日

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  • Motahare-Sadat Hosseini,
  • Fereshteh Barjesteh,
  • Fereshteh Azedi,
  • Iraj Alipourfard,
  • Zahra Rezaei,
  • Elham Bahreini

摘要

Introduction

The higher prevalence of non-alcoholic fatty liver disease (NAFLD) in men than women before menopause and the reduced difference post-menopause suggest that sex hormones may influence liver lipid accumulation. This study compared the effects of sex hormones on lipid droplet (LD) accumulation in palmitate/ oleate-treated HepG2 cells.

Methods

The MTT method was used to determine effective doses of palmitic and oleic acids in HepG2 cells, followed by a combined dose for inducing LD formation. Changes in LD content after treatment with various doses of β-estradiol and testosterone were evaluated qualitatively and semi-quantitatively using Oil Red O staining and light microscopy. The effects of these hormones on gene expression related to LD formation and lipogenesis, including PLIN2, ATGL, CGI-58, and CIDEB, were assessed using quantitative PCR.

Results

Treatment of HepG2 cells with palmitate and oleate increased LD accumulation and the expression of PLIN2 and CIDE while elevating ATGL expression without affecting CGI-58. With no significant difference, both β-estradiol and testosterone significantly reduced LD accumulation in steatotic HepG2 cells. Gene analysis indicated that both hormones decreased PLIN2 and increased CGI-58 expression. Testosterone did not affect CIDE, while β-estradiol reduced it at low doses. Combined treatment showed no significant changes in gene expression compared to individual hormone effects, but LD accumulation was synergistically reduced.

Conclusion

This study demonstrates that β-estradiol and testosterone significantly modulate LD content and the expression of key regulatory genes in HepG2 cells, with β-estradiol showing a somewhat dominant role in enhancing lipid turnover and mitigating lipid accumulation.