25-Hydroxycholesterol Inhibits Adipogenic Differentiation of C3H10T1/2 Pluripotent Stromal Cells.

Chongxiao Chen,Woo Kyun Kim,Alemu Regassa,Karmin O,Dorothy Moseti

doi:10.3390/ijms21020412

Abstract

Understanding of adipogenesis is important to find remedies for obesity and related disorders. In addition, it is also critical in bone disorders because there is a reciprocal relationship between adipogenesis and osteogenesis in bone micro-environment. Oxysterols are pro-osteogenic and anti-adipogenic molecules via hedgehog activation in pluripotent bone marrow stomal cells. However, no study has evaluated the role of specific oxysterols in C3H10T1/2 cells, which are a good cell model for studying osteogenesis and adipogenesis in bone-marrows. Thus, we investigated the effects of specific oxysterols on adipogenesis and expression of adipogenic transcripts in C3H10T1/2 cells. Treatment of cells with DMITro significantly induced mRNA expression of Pparγ. This induction was significantly inhibited by 25-HC. The expression of C/cepα, Fabp4 and Lpl was also inhibited by 25-HC. To determine the mechanism by which 25-HC inhibits adipogenesis, the effects of the hedgehog signalling pathway inhibitor, cyclopamine and CUR61414, were evaluated. Treatment of C3H10T1/2 cells with DMITro + cyclopamine or DMITro + CUR61414 for 96h did not modulate adipocyte differentiation; cyclopamine and CUR61414 did not reverse the inhibitory effects of 25-HC, suggesting that the canonical hedgehog signalling may not play a role in the anti-adipogenic effects of 25-HC in C3H10T1/2 cells. In addition, LXR agonist did not inhibit adipogenesis, but 25-HC strongly inhibits adipogenesis of C3H10T1/2 cells. Our observations showed that 25-HC was the most potent oxysterol in inhibiting adipogenesis and the expression of key adipogenic transcripts in C3H10T1/2 cells among the tested oxysterols, suggesting its potential application in providing an intervention in osteoporosis and obesity. We also report that the inhibitory effects of 25-HC on adipogenic differentiation in C3H10T1/2 cells are not mediated by hedgehog signaling and LXR.

Highlights

Obesity is a major health problem that leads to increased risk of type II diabetes mellitus (T2DM), cardiovascular diseases and hypertension and has been associated with high morbidity and mortality rates, especially in the industrialized world [1]
We demonstrated that 25-HC inhibits adipogenic differentiation of C3H10T1/2 mouse stromal cells
Of the four oxysterols tested, 25-HC proved to be the most potent in inhibiting accumulation of cyctoplasmic lipid droplets and expression of adipogenic protein markers in the cells. 25-HC inhibited the expression of Pparγ, which is the main regulator of adipogenesis

Summary

Introduction

Obesity is a major health problem that leads to increased risk of type II diabetes mellitus (T2DM), cardiovascular diseases and hypertension and has been associated with high morbidity and mortality rates, especially in the industrialized world [1]. Obesity is associated with adipose tissue and the development of fat cells or adipocytes [2,3]. Adipocytes play an important role in maintaining energy balance in the body of animals [6]. Excess energy intake leads to an increase in the adipose tissue due to an increase in the number or size of adipocytes, which leads to obesity [8]. Excess body fats lead to accumulation of cholesterol in arterial wall leading to atherosclerosis, a major cardiovascular disease. The enlarged fat cells secrete certain adipokines, which hinder insulin signalling leading to development of T2DM [7]

Methods

Results

Conclusion