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miR-125a-3p and miR-483-5p promote adipogenesis via suppressing the RhoA/ROCK1/ERK1/2 pathway in multiple symmetric lipomatosis.

Chen K, He H, Xie Y, Zhao L, Zhao S, Wan X, Yang W, Mo Z - Sci Rep (2015)

Bottom Line: Most importantly, we found that miR-125a-3p and miR-483-5p promoted de novo adipose tissue formation in nude mice.These results demonstrated that miR-125a-3p and miR-483-5p coordinately promoted adipogenesis through suppressing the RhoA/ROCK1/ERK1/2 pathway.Our findings may provide novel strategies for the management and treatment of MSL or obesity.

View Article: PubMed Central - PubMed

Affiliation: Department of Endocrinology, Third Xiangya Hospital of Central South University, Changsha, Hunan, 410013, China.

ABSTRACT
Multiple symmetric lipomatosis (MSL) is a rare disease characterized by symmetric and abnormal distribution of subcutaneous adipose tissue (SAT); however, the etiology is largely unknown. We report here that miR-125a-3p and miR-483-5p are upregulated in the SAT of MSL patients, promoting adipogenesis through suppressing the RhoA/ROCK1/ERK1/2 pathway. TaqMan microRNA (miR) array analysis revealed that 18 miRs were upregulated in the SAT of MSL patients. Transfection of human adipose-derived mesenchymal stem cells (hADSCs) with the individual agomirs of these 18 miRs showed that miR-125a-3p and miR-483-5p significantly promoted adipogenesis. A dual-luciferase assay showed that RhoA and ERK1 were the targets of miR-125a-3p and miR-483-5p, respectively. Moreover, transfection of hADSCs with mimics of miR-125a-3p and miR-483-5p resulted in a pronounced decrease of ERK1/2 phosphorylation in the nucleus; conversely, transfection of hADSCs with inhibitors of miR-125a-3p and miR-483-5p led to a significant increase of ERK1/2 phosphorylation in the nucleus. Most importantly, we found that miR-125a-3p and miR-483-5p promoted de novo adipose tissue formation in nude mice. These results demonstrated that miR-125a-3p and miR-483-5p coordinately promoted adipogenesis through suppressing the RhoA/ROCK1/ERK1/2 pathway. Our findings may provide novel strategies for the management and treatment of MSL or obesity.

No MeSH data available.


Related in: MedlinePlus

miR-125a-3p and miR-483-5p regulate adipogenesis in hADSCs.hADSCs were isolated from the abdominal SAT of two control patient respectively, and we repeated all experiments by two kinds of hADSCs. The hADSCs were transfected with the miR-125a-3p or miR-483-5p agomir, agomir-NC (100 nM), antagomir-NC, or antagomir (200 nM), respectively. After 72 h, (A) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (B) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed by a camera (no magnification). (C) The hADSCs underwent self-differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (D,E) The hADSCs were induced differentiation for 12 days, the protein levels of PPARγ, C/EBPα, and FABP4 were detected by Western blot. β-Actin served as a loading control. All measurements were preformed by three independent experiments.
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f2: miR-125a-3p and miR-483-5p regulate adipogenesis in hADSCs.hADSCs were isolated from the abdominal SAT of two control patient respectively, and we repeated all experiments by two kinds of hADSCs. The hADSCs were transfected with the miR-125a-3p or miR-483-5p agomir, agomir-NC (100 nM), antagomir-NC, or antagomir (200 nM), respectively. After 72 h, (A) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (B) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed by a camera (no magnification). (C) The hADSCs underwent self-differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (D,E) The hADSCs were induced differentiation for 12 days, the protein levels of PPARγ, C/EBPα, and FABP4 were detected by Western blot. β-Actin served as a loading control. All measurements were preformed by three independent experiments.

Mentions: To investigate which miRs may promote adipogenesis, all 18 miRs with agomir were respectively transfected into hADSCs, which were further induced to mature adipocytes or underwent self-differentiation for 12 days. We found that transfection with miR-125a-3p or miR-483-5p significantly promoted lipid droplet accumulation in hADSCs that were induced to mature adipocytes. Conversely, inhibition of miR-125a-3p or miR-483-5p with the corresponding antagomir markedly decreased lipid droplet accumulation (Fig. 2A,B). The same results were observed in the hADSCs undergoing self-differentiation for 12 days (Fig. 2C). Next, after each group was induced to mature adipocytes for 12 days, C/EBPα, PPARγ, and FABP4 were upregulated by the agomir of miR-125a-3p or miR-483-5p; while they were downregulated by the antagomir of miR-125a-3p or miR-483-5p (Fig. 2D,E). These results demonstrated that miR-125a-3p and miR-483-5p promote adipogenesis in hADSCs.


miR-125a-3p and miR-483-5p promote adipogenesis via suppressing the RhoA/ROCK1/ERK1/2 pathway in multiple symmetric lipomatosis.

Chen K, He H, Xie Y, Zhao L, Zhao S, Wan X, Yang W, Mo Z - Sci Rep (2015)

miR-125a-3p and miR-483-5p regulate adipogenesis in hADSCs.hADSCs were isolated from the abdominal SAT of two control patient respectively, and we repeated all experiments by two kinds of hADSCs. The hADSCs were transfected with the miR-125a-3p or miR-483-5p agomir, agomir-NC (100 nM), antagomir-NC, or antagomir (200 nM), respectively. After 72 h, (A) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (B) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed by a camera (no magnification). (C) The hADSCs underwent self-differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (D,E) The hADSCs were induced differentiation for 12 days, the protein levels of PPARγ, C/EBPα, and FABP4 were detected by Western blot. β-Actin served as a loading control. All measurements were preformed by three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493643&req=5

f2: miR-125a-3p and miR-483-5p regulate adipogenesis in hADSCs.hADSCs were isolated from the abdominal SAT of two control patient respectively, and we repeated all experiments by two kinds of hADSCs. The hADSCs were transfected with the miR-125a-3p or miR-483-5p agomir, agomir-NC (100 nM), antagomir-NC, or antagomir (200 nM), respectively. After 72 h, (A) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (B) The hADSCs were induced differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed by a camera (no magnification). (C) The hADSCs underwent self-differentiation for 12 days, lipid droplet accumulation stained by Oil red O was observed under a microscope. (D,E) The hADSCs were induced differentiation for 12 days, the protein levels of PPARγ, C/EBPα, and FABP4 were detected by Western blot. β-Actin served as a loading control. All measurements were preformed by three independent experiments.
Mentions: To investigate which miRs may promote adipogenesis, all 18 miRs with agomir were respectively transfected into hADSCs, which were further induced to mature adipocytes or underwent self-differentiation for 12 days. We found that transfection with miR-125a-3p or miR-483-5p significantly promoted lipid droplet accumulation in hADSCs that were induced to mature adipocytes. Conversely, inhibition of miR-125a-3p or miR-483-5p with the corresponding antagomir markedly decreased lipid droplet accumulation (Fig. 2A,B). The same results were observed in the hADSCs undergoing self-differentiation for 12 days (Fig. 2C). Next, after each group was induced to mature adipocytes for 12 days, C/EBPα, PPARγ, and FABP4 were upregulated by the agomir of miR-125a-3p or miR-483-5p; while they were downregulated by the antagomir of miR-125a-3p or miR-483-5p (Fig. 2D,E). These results demonstrated that miR-125a-3p and miR-483-5p promote adipogenesis in hADSCs.

Bottom Line: Most importantly, we found that miR-125a-3p and miR-483-5p promoted de novo adipose tissue formation in nude mice.These results demonstrated that miR-125a-3p and miR-483-5p coordinately promoted adipogenesis through suppressing the RhoA/ROCK1/ERK1/2 pathway.Our findings may provide novel strategies for the management and treatment of MSL or obesity.

View Article: PubMed Central - PubMed

Affiliation: Department of Endocrinology, Third Xiangya Hospital of Central South University, Changsha, Hunan, 410013, China.

ABSTRACT
Multiple symmetric lipomatosis (MSL) is a rare disease characterized by symmetric and abnormal distribution of subcutaneous adipose tissue (SAT); however, the etiology is largely unknown. We report here that miR-125a-3p and miR-483-5p are upregulated in the SAT of MSL patients, promoting adipogenesis through suppressing the RhoA/ROCK1/ERK1/2 pathway. TaqMan microRNA (miR) array analysis revealed that 18 miRs were upregulated in the SAT of MSL patients. Transfection of human adipose-derived mesenchymal stem cells (hADSCs) with the individual agomirs of these 18 miRs showed that miR-125a-3p and miR-483-5p significantly promoted adipogenesis. A dual-luciferase assay showed that RhoA and ERK1 were the targets of miR-125a-3p and miR-483-5p, respectively. Moreover, transfection of hADSCs with mimics of miR-125a-3p and miR-483-5p resulted in a pronounced decrease of ERK1/2 phosphorylation in the nucleus; conversely, transfection of hADSCs with inhibitors of miR-125a-3p and miR-483-5p led to a significant increase of ERK1/2 phosphorylation in the nucleus. Most importantly, we found that miR-125a-3p and miR-483-5p promoted de novo adipose tissue formation in nude mice. These results demonstrated that miR-125a-3p and miR-483-5p coordinately promoted adipogenesis through suppressing the RhoA/ROCK1/ERK1/2 pathway. Our findings may provide novel strategies for the management and treatment of MSL or obesity.

No MeSH data available.


Related in: MedlinePlus