Insulin-stimulated lipid accumulation is inhibited by ROS-scavenging chemicals, but not by the Drp1 inhibitor Mdivi-1

PLoS One. 2017 Oct 2;12(10):e0185764. doi: 10.1371/journal.pone.0185764. eCollection 2017.

Abstract

Adipocyte differentiation is regulated by intracellular reactive oxygen species (ROS) generation and mitochondrial fission and fusion processes. However, the correlation between intracellular ROS generation and mitochondrial remodeling during adipocyte differentiation is still unknown. Here, we investigated the effect on adipocyte differentiation of 3T3-L1 cells of intracellular ROS inhibition using N-acetyl cysteine (Nac) and Mito-TEMPO and of mitochondrial fission inhibition using Mdivi-1. Differentiated 3T3-L1 adipocytes displayed an increase in mitochondrial fission, ROS generation, and the expression of adipogenic and mitochondrial dynamics-related proteins. ROS scavenger (Nac or Mito-TEMPO) treatment inhibited ROS production, lipid accumulation, the expression of adipogenic and mitochondrial dynamics-related proteins, and mitochondrial fission during adipogenesis of 3T3-L1 cells. On the other hand, treatment with the mitochondrial fission inhibitor Mdivi-1 inhibited mitochondrial fission but did not inhibit ROS production, lipid accumulation, or the expression of adipogenic and mitochondrial dynamics-related proteins, with the exception of phosphorylated Drp1 (Ser616), in differentiated 3T3-L1 adipocytes. The inhibition of mitochondrial fission did not affect adipocyte differentiation, while intracellular ROS production decreased in parallel with inhibition of adipocyte differentiation. Therefore, our results indicated that ROS are an essential regulator of adipocyte differentiation in 3T3-L1 cells.

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / cytology
  • Adipocytes / metabolism
  • Animals
  • Antioxidants / metabolism
  • Dynamins / antagonists & inhibitors*
  • Free Radicals / metabolism*
  • Insulin / pharmacology*
  • Lipid Metabolism*
  • Mice
  • Quinazolinones / pharmacology*
  • Reactive Oxygen Species / metabolism*

Substances

  • 3-(2,4-dichloro-5-methoxyphenyl)-2-sulfanyl-4(3H)-quinazolinone
  • Antioxidants
  • Free Radicals
  • Insulin
  • Quinazolinones
  • Reactive Oxygen Species
  • Dnm1l protein, mouse
  • Dynamins

Grants and funding

This research was supported by grants from the KRIBB Research Initiative Program (KGM4611714), the Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01179202), and the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2017R1A2B4008176 and NRF-2014R1A2A1A11054095). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.