Endocrinol Metab.  2016 Jun;31(2):328-335. 10.3803/EnM.2016.31.2.328.

The Effects of High Fat Diet and Resveratrol on Mitochondrial Activity of Brown Adipocytes

• Ku CR
• Cho YH
• Hong ZY
• Lee H
• Lee SJ
• Hong SS
• Lee EJ

• KMID: 2308864
• DOI: http://doi.org/10.3803/EnM.2016.31.2.328

Abstract

BACKGROUND
Resveratrol (RSV) is a polyphenolic phytoalexin that has many effects on metabolic diseases such as diabetes and obesity. Given the importance of brown adipose tissue (BAT) for energy expenditure, we investigated the effects of RSV on brown adipocytes.
METHODS
For the in vitro study, interscapular BAT was isolated from 7-week-old male Sprague Dawley rats. For the in vivo study, 7-week-old male Otsuka Long Evans Tokushima Fatty (OLETF) rats were divided into four groups and treated for 27 weeks with: standard diet (SD); SD+RSV (10 mg/kg body weight, daily); high fat diet (HFD); HFD+RSV. RSV was provided via oral gavage once daily during the in vivo experiments.
RESULTS
RSV treatment of primary cultured brown preadipocytes promoted mitochondrial activity, along with over-expression of estrogen receptor α (ER-α). In OLETF rats, both HFD and RSV treatment increased the weight of BAT and the differentiation of BAT. However, only RSV increased the mitochondrial activity and ER-α expression of BAT in the HFD-fed group. Finally, RSV improved the insulin sensitivity of OLETF rats by increasing the mitochondrial activity of BAT, despite having no effects on white adipocytes and muscles in either diet group.
CONCLUSION
RSV could improve insulin resistance, which might be associated with mitochondrial activity of brown adipocyte. Further studies evaluating the activity of RSV for both the differentiation and mitochondrial activity of BAT could be helpful in investigating the effects of RSV on metabolic parameters.

Keyword

Adipocytes, brown; Estrogen receptor alpha; Diet, high-fat; Mitochondria; Resveratrol

MeSH Terms

Adipocytes, Brown*
Adipocytes, White
Adipose Tissue, Brown
Animals
Body Weight
Diet
Diet, High-Fat*
Energy Metabolism
Estrogen Receptor alpha
Estrogens
Humans
In Vitro Techniques
Insulin Resistance
Male
Metabolic Diseases
Mitochondria
Muscles
Obesity
Rats
Rats, Inbred OLETF
Rats, Sprague-Dawley
Estrogen Receptor alpha
Estrogens

Figure

• 

  Fig. 1 Effects of resveratrol (RSV) on the differentiation of brown adipocytes. (A) Oil-red-O staining of primary cultured brown preadipocytes. RSV treatment was performed with induction medium at doses of 50 and 100 µM and maintained thereafter. Adipogenesis was significantly increased with RSV treatment. (B) Western blotting for mitochondrial activity and estrogen receptor α (ER-α) in brown adipocytes. Brown preadipocytes were cultured and differentiated with or without RSV treatment. The protein lysates were collected after the differentiation of brown adipocytes was confirmed. UCP-1, uncoupling protein 1; p-AMPK, phospho-AMP-activated protein kinase; AMPK, AMP-activated protein kinase.

• 

  Fig. 2 Effects of resveratrol (RSV) on metabolic parameters in Otsuka Long Evans Tokushima Fatty (OLETF) rats. (A) Body weight changes after RSV treatment of OLETF rats. RSV had been provided since the age of 9 weeks and maintained for 27 weeks. (B) Insulin tolerance test of OLETF rats at 35 weeks of age. (C) Organ weights at 36 weeks of age. (D) Changes in leptin and adiponectin levels. Serum samples from 36-week-old rats were collected and evaluated with an enzyme-linked immunoassay kit. SD, standard diet; HF, high fat; AUC, area under the curve. aP value below 0.05 with treatment of RSV in each diet group (Mann-Whitney U test).

• 

  Fig. 3 Effects of resveratrol (RSV) on the differentiation and activation of brown adipocytes in Otsuka Long Evans Tokushima Fatty (OLETF) rats. (A) Hematoxylin and eosin staining for brown adipocytes. Brown adipose tissue was collected from OLETF rats at 36 weeks of age. With the treatment of RSV and feeding of the high fat diet (HFD), the differentiation of brown adipocytes increased. (B) The number of brown adipocytes visible in the high-power field increased significantly with RSV treatment and HFD feeding. The average number of brown adipocytes was calculated at four sites of each section. The number of rats in each group is described in the Methods. (C) Western blotting for mitochondrial activity and estrogen receptor α (ER-α). (D) Relative expression in Western blotting, determined using a densitometer. All protein lysates were evaluated and compared with those of rats on the standard diet. SD, standard diet; HF, high fat; UCP-1, uncoupling protein 1; p-AMPK, phospho-AMP-activated protein kinase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. aP value below 0.05 with treatment of RSV in each diet group (Mann-Whitney U test).

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