Korean Diabetes J.  2009 Jun;33(3):206-214. 10.4093/kdj.2009.33.3.206.

Effect of Adipose Differentiation-Related Protein (ADRP) on Glucose Uptake of Skeletal Muscle

• Ku YH
• Kim M
• Kim S
• Park HS
• Kim HJ
• Lee IK
• Shin DH
• Chung SS
• Park SG
• Cho YM
• Lee HK
• Park KS

• KMID: 2222420
• DOI: http://doi.org/10.4093/kdj.2009.33.3.206

Abstract

BACKGROUND
Skeletal muscle is the most important tissue contributing to insulin resistance. Several studies have shown that accumulation of intramyocellular lipid is associated with the development of insulin resistance. Thus, proteins involved in lipid transport, storage and metabolism might also be involved in insulin action in skeletal muscle. Adipose differentiation-related protein (ADRP), which is localized at the surface of lipid droplets, is known to be regulated by peroxisome proliferator activated receptor gamma (PPARgamma). However, it is not known whether ADRP plays a role in regulating glucose uptake and insulin action in skeletal muscle. METHODS: ADRP expression in skeletal muscle was measured by RT-PCR and western blot in db/db mice with and without PPARgamma agonist. The effect of PPARgamma agonist or high lipid concentration (0.4% intralipos) on ADRP expression was also obtained in cultured human skeletal muscle cells. Glucose uptake was measured when ADRP was down-regulated with siRNA or when ADRP was overexpressed with adenovirus. RESULTS: ADRP expression increased in the skeletal muscle of db/db mice in comparison with normal controls and tended to increase with the treatment of PPARgamma agonist. In cultured human skeletal muscle cells, the treatment of PPARgamma agonist or high lipid concentration increased ADRP expression. siADRP treatment decreased both basal and insulin-stimulated glucose uptake whereas ADRP overexpression increased glucose uptake in cultured human skeletal muscle cells. CONCLUSION: ADRP expression in skeletal muscle is increased by PPARgamma agonist or exposure to high lipid concentration. In these conditions, increased ADRP contributed to increase glucose uptake. These results suggest that insulin-sensitizing effects of PPARgamma are at least partially achieved by the increase of ADRP expression, and ADRP has a protective effect against intramyocellular lipid-induced insulin resistance.

Keyword

Adipose differentiation related protein (ADRP); Insulin resistance; Peroxisome proliferator activated receptor gamma; Skeletal muscle

MeSH Terms

Adenoviridae
Animals
Blotting, Western
Glucose
Humans
Insulin
Insulin Resistance
Membrane Proteins
Mice
Muscle, Skeletal
PPAR gamma
Proteins
RNA, Small Interfering
Glucose
Insulin
Membrane Proteins
PPAR gamma
Proteins
RNA, Small Interfering

Figure

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  Fig. 1 Effect of rosiglitazone treatment on blood glucose, body weight and triglyceride (TG) in db/db mice. A. Blood glucose level. B. Body weight. C. Food intake. D. Water intake. E. Fasting plasma glucose. F. Fasting TG level at 14 days after rosiglitazone treatment. db/db + rosi: db/db + rosiglitazone treatment, *P < 0.01 vs. C57BL6J, †P < 0.01 vs. db/db mice.

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  Fig. 2 Effect of rosiglitazone on skeletal muscle ADRP expression in db/db mice. A. RT-PCR. B. Western blotting. db/db + rosi: db/db + rosiglitazone treatment. *P < 0.01 vs. C57BL6.

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  Fig. 3 Effect of PPARγ agonists on ADRP expression in cultured human skeletal muscle cells. Human skeletal muscle cells were differentiated and treated with 10 µM of troglitazone, rosiglitazone, pioglitazone, or wy14,643 for 48 h. *P < 0.01 vs. vehicle only.

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  Fig. 4 Effect of ADRP knockdown on glucose uptake. Human skeletal muscle cells (HSkMC) were transfected with siRNAs of negative control (siNS) or ADRP (siADRP) for 12 h and treated with troglitazone for an additional 48 h. Prior to perform glucose uptake assay, cells were incubated with or without insulin (100 nM) for 30 min. (A) siADRP decreased both mRNA and protein expression of ADRP in HSkMC (B) glucose uptake in HSkMC. *P < 0.05: vs. basal value of control cells not treated with troglitazone; †P < 0.05: vs. corresponding value of control cells not treated with troglitazone, ‡P < 0.05 vs. corresponding value of cells treated with troglitazone and siNS.

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  Fig. 5 Effect of ADRP overexpression on glucose uptake. A. Human skeletal muscle cells were transfected with adenovirus which expresses ADRP for 36 h. The increase in ADRP protein was determined by western blot. B. Glucose uptake in HSkMC. *P < 0.05 vs. the basal value of GFP adenovirus treated cells. †P < 0.05 vs. the corresponding value of GFP adenovirus transfected cells.

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  Fig. 6 Role of ADRP on glucose uptake in cultured human skeletal muscle cells exposed to high lipid concentration A. High lipid (intralipos 400 mg/dL) induces ADRP mRNA expression in HSkMC. B. Human skeletal muscle cells were transfected with siNS or siADRP for 12 h and treated with high lipid for 48 h. siADRP inhibited high lipid-induced ADRP protein expression. C. Glucose uptake in siADRP transfected HSkMC with or without high lipid. *P < 0.05: vs. basal value of siNS treated cells. †P < 0.05: vs. corresponding value of siNS transfected cells not treated with high lipid, ‡P < 0.05: vs. corresponding value of cells treated with siNS and high lipid.

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