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  • In vitro exploration of ACAT contributions to lipid droplet formation during adipogenesis.

In vitro exploration of ACAT contributions to lipid droplet formation during adipogenesis.

Journal of lipid research (2018-03-20)
Yuyan Zhu, Chih-Yu Chen, Junjie Li, Ji-Xin Cheng, Miran Jang, Kee-Hong Kim
ABSTRACT

As adipose tissue is the major cholesterol storage organ and most of the intracellular cholesterol is distributed to lipid droplets (LDs), cholesterol homeostasis may have a role in the regulation of adipocyte size and function. ACATs catalyze the formation of cholesteryl ester (CE) from free cholesterol to modulate the cholesterol balance. Despite the well-documented role of ACATs in hypercholesterolemia, their role in LD development during adipogenesis remains elusive. Here, we identify ACATs as regulators of de novo lipogenesis and LD formation in murine 3T3-L1 adipocytes. Pharmacological inhibition of ACAT activity suppressed intracellular cholesterol and CE levels, and reduced expression of genes involved in cholesterol uptake and efflux. ACAT inhibition resulted in decreased de novo lipogenesis, as demonstrated by reduced maturation of SREBP1 and SREBP1-downstream lipogenic gene expression. Consistent with this observation, knockdown of either ACAT isoform reduced total adipocyte lipid content by approximately 40%. These results demonstrate that ACATs are required for storage ability of lipids and cholesterol in adipocytes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glycerol Standard Solution, 2.5 mg/ml equivalent triolein concentration
Sigma-Aldrich
Free Glycerol Reagent, used for quantitative enzymatic determination of glycerol
Sigma-Aldrich
Triglyceride Reagent, used for quantitative enzymatic determination of triglyerides