Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes
ObjectiveGhrelin is a 28-amino-acid peptide that regulates energy hemostasis, glucose and lipid metabolism. We aimed to explore the effects of ghrelin on the proliferation and differentiation of 3T3-L1 and human primary preadipocytes.Methods3-(4,5-Dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromi...
Main Authors: | , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2019-10-01
|
Series: | Frontiers in Physiology |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2019.01296/full |
id |
doaj-4c0f876164d5436d8ce5c5dfec6388f2 |
---|---|
record_format |
Article |
spelling |
doaj-4c0f876164d5436d8ce5c5dfec6388f22020-11-25T01:15:25ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2019-10-011010.3389/fphys.2019.01296472349Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary PreadipocytesHui MiaoHui PanLinjie WangHongbo YangHuijuan ZhuFengying GongObjectiveGhrelin is a 28-amino-acid peptide that regulates energy hemostasis, glucose and lipid metabolism. We aimed to explore the effects of ghrelin on the proliferation and differentiation of 3T3-L1 and human primary preadipocytes.Methods3-(4,5-Dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) spectrophotometry, Oil Red O staining, intracellular glycerol-3-phosphate dehydrogenase (G-3-PDH) assays and semiquantitative reverse transcription polymerase chain reaction were used to investigate the action of ghrelin.ResultsGhrelin (0.01–1000 ng/ml) significantly increased the numbers of 3T3-L1 cells, and the maximum stimulatory effect was observed with the 10 ng/ml ghrelin treatment for 24 h (p < 0.05). Ghrelin also promoted the proliferation of human primary preadipocytes from 24 h (p < 0.05) to 48 h (p < 0.05) at a concentration of 1000 ng/ml. Further investigation showed that IGF-1 levels were notably increased in ghrelin-treated 3T3-L1 and human preadipocytes, and IGF-1 antibody was capable to attenuate this stimulatory action of ghrelin (all p < 0.05). Additionally, ghrelin significantly suppressed the differentiation of 3T3-L1 and human primary preadipocytes; 10 ng/ml ghrelin notably downregulated G-3-PDH activities in 3T3-L1 cells on day 3 and in human cells from days 4 to 12 following differentiation (all p < 0.05), and the intracellular lipoprotein lipase mRNA levels were lower than that of the controls (p < 0.05). Further investigation showed that the mRNA levels of peroxisome proliferator-activated receptor γ2 (PPARγ2) and CCAAT/enhancer binding protein α (C/EBPs) were also suppressed in ghrelin-treated human differentiating adipocytes.ConclusionGhrelin promotes the proliferation of 3T3-L1 and human primary preadipocytes by increasing the expression of IGF-1. Ghrelin inhibits murine and human adipocyte differentiation by downregulating PPARγ2 and C/EBPα levels, consequently leading to decreased lipid accumulation and lipogenic enzymes expression.https://www.frontiersin.org/article/10.3389/fphys.2019.01296/fullghrelinproliferationdifferentiation3T3-L1 preadipocyteshuman primary preadipocytes |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hui Miao Hui Pan Linjie Wang Hongbo Yang Huijuan Zhu Fengying Gong |
spellingShingle |
Hui Miao Hui Pan Linjie Wang Hongbo Yang Huijuan Zhu Fengying Gong Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes Frontiers in Physiology ghrelin proliferation differentiation 3T3-L1 preadipocytes human primary preadipocytes |
author_facet |
Hui Miao Hui Pan Linjie Wang Hongbo Yang Huijuan Zhu Fengying Gong |
author_sort |
Hui Miao |
title |
Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes |
title_short |
Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes |
title_full |
Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes |
title_fullStr |
Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes |
title_full_unstemmed |
Ghrelin Promotes Proliferation and Inhibits Differentiation of 3T3-L1 and Human Primary Preadipocytes |
title_sort |
ghrelin promotes proliferation and inhibits differentiation of 3t3-l1 and human primary preadipocytes |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Physiology |
issn |
1664-042X |
publishDate |
2019-10-01 |
description |
ObjectiveGhrelin is a 28-amino-acid peptide that regulates energy hemostasis, glucose and lipid metabolism. We aimed to explore the effects of ghrelin on the proliferation and differentiation of 3T3-L1 and human primary preadipocytes.Methods3-(4,5-Dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) spectrophotometry, Oil Red O staining, intracellular glycerol-3-phosphate dehydrogenase (G-3-PDH) assays and semiquantitative reverse transcription polymerase chain reaction were used to investigate the action of ghrelin.ResultsGhrelin (0.01–1000 ng/ml) significantly increased the numbers of 3T3-L1 cells, and the maximum stimulatory effect was observed with the 10 ng/ml ghrelin treatment for 24 h (p < 0.05). Ghrelin also promoted the proliferation of human primary preadipocytes from 24 h (p < 0.05) to 48 h (p < 0.05) at a concentration of 1000 ng/ml. Further investigation showed that IGF-1 levels were notably increased in ghrelin-treated 3T3-L1 and human preadipocytes, and IGF-1 antibody was capable to attenuate this stimulatory action of ghrelin (all p < 0.05). Additionally, ghrelin significantly suppressed the differentiation of 3T3-L1 and human primary preadipocytes; 10 ng/ml ghrelin notably downregulated G-3-PDH activities in 3T3-L1 cells on day 3 and in human cells from days 4 to 12 following differentiation (all p < 0.05), and the intracellular lipoprotein lipase mRNA levels were lower than that of the controls (p < 0.05). Further investigation showed that the mRNA levels of peroxisome proliferator-activated receptor γ2 (PPARγ2) and CCAAT/enhancer binding protein α (C/EBPs) were also suppressed in ghrelin-treated human differentiating adipocytes.ConclusionGhrelin promotes the proliferation of 3T3-L1 and human primary preadipocytes by increasing the expression of IGF-1. Ghrelin inhibits murine and human adipocyte differentiation by downregulating PPARγ2 and C/EBPα levels, consequently leading to decreased lipid accumulation and lipogenic enzymes expression. |
topic |
ghrelin proliferation differentiation 3T3-L1 preadipocytes human primary preadipocytes |
url |
https://www.frontiersin.org/article/10.3389/fphys.2019.01296/full |
work_keys_str_mv |
AT huimiao ghrelinpromotesproliferationandinhibitsdifferentiationof3t3l1andhumanprimarypreadipocytes AT huipan ghrelinpromotesproliferationandinhibitsdifferentiationof3t3l1andhumanprimarypreadipocytes AT linjiewang ghrelinpromotesproliferationandinhibitsdifferentiationof3t3l1andhumanprimarypreadipocytes AT hongboyang ghrelinpromotesproliferationandinhibitsdifferentiationof3t3l1andhumanprimarypreadipocytes AT huijuanzhu ghrelinpromotesproliferationandinhibitsdifferentiationof3t3l1andhumanprimarypreadipocytes AT fengyinggong ghrelinpromotesproliferationandinhibitsdifferentiationof3t3l1andhumanprimarypreadipocytes |
_version_ |
1725153391462580224 |