Effect of dietary resveratrol on the metabolic profile of nutrients in obese OLETF rats
© Nagao et al.; licensee BioMed Central Ltd. 2013
Received: 23 December 2012
Accepted: 31 January 2013
Published: 4 February 2013
Resveratrol (trans-3,4′,5-trihydroxystilbene) is a naturally occurring phytoalexin produced by plants in response to various stresses. Several studies have shown that resveratrol is present in significant amounts in a variety of human diets, including wines, grapes, berries, and peanuts, and it possesses several beneficial health properties, such as atheroprotective, anti-obesity, anti-cancer, anti-inflammatory and antioxidant activities. In this study, we evaluated the effect of resveratrol on the pathogenesis of obesity and the metabolic profile of nutrients in non-high fat-fed obese OLETF rats.
Although lipid parameters in the serum and liver were not changed, the accumulation of abdominal white adipose tissues was markedly prevented in resveratrol diet-fed OLETF rats after 4 weeks of feeding. The results of the respiratory gas analysis indicated that dietary resveratrol induced the partial enhancement of fat metabolism and sparing actions for carbohydrate and protein at 1 week and 3 weeks of feeding in OLETF rats. Additionally, the adipose mRNA level of carnitine palmitoyltransferase in the resveratrol diet-fed OLETF rats was higher than the control rats after 4 weeks of feeding.
Our study demonstrated that dietary resveratrol can prevent obesity through a change in the metabolic profile of nutrients in obese OLETF rats.
KeywordsResveratrol Respiratory gas analysis Metabolic profile Obesity OLETF rat
Lifestyle-related diseases, such as hyperlipidemia, arteriosclerosis, diabetes mellitus, and hypertension, are widespread and increasingly prevalent in industrialized countries and contribute to the increase in cardiovascular morbidity and mortality [1, 2]. Accompanied by the rapid increase in the number of elderly people, they are important not only medically but also socioeconomically. Although the pathogenesis of lifestyle-related diseases is complicated and the precise mechanisms have not been elucidated, obesity has emerged as one of the major cardiovascular risk factors according to epidemiologic studies [3–5]. Obesity is defined as an increased mass of adipose tissue, and its prevalence and severity are markedly increasing in Westernized countries.
Diet has been recognized as a factor that contributes to the development and prevention of lifestyle-related diseases, and various natural molecules in plants and fruits have been used in folk medicines throughout the world for treating those diseases [6–10]. Resveratrol (trans-3,4′,5-trihydroxystilbene) is a naturally occurring phytoalexin produced by plants in response to various stresses. Several studies have shown that resveratrol is present in significant amounts in a variety of human diets, including wines, grapes, berries, and peanuts, and it possesses several beneficial health properties, such as atheroprotective, anti-cancer, anti-inflammatory and antioxidant activities [11–13]. Recent studies have also reported that resveratrol has anti-obesity effects in high-fat-fed mice and rats [14, 15]. However, the effects of resveratrol on the metabolic profile of nutrients and body fat accumulation in non-high-fat diet-fed rodents have not been fully evaluated. Otsuka Long-Evans Tokushima fatty (OLETF) rats develop a syndrome with multiple metabolic and hormonal disorders that shares many features with human obesity [16–18]. OLETF rats have hyperphagia because they lack receptors for cholecystokinin, and they become obese even by consuming a normal diet (AIN-76, 7% fat diet) [19–21]. Therefore, we evaluated the effect of resveratrol on the pathogenesis of obesity and the metabolism of carbohydrate, fat, and protein in obese OLETF rats.
Materials and methods
Animals and diets
The composition of the experimental diets
Mineral mixture (AIN-76)
Vitamin mixture (AIN-76)
Measurement of the triglyceride, cholesterol and glycogen levels in the liver
The liver lipids were extracted, and the concentrations of triglyceride and cholesterol were determined as described elsewhere . The glycogen levels were determined according to the method of Lo et al. .
Measurement of the serum parameters
The serum triglyceride, cholesterol, and glucose levels were measured using commercial enzyme assay kits (Wako Pure Chemicals, Tokyo, Japan).
Respiratory gas analysis
The instruments and software used for the measurement of the oxygen consumption and respiratory quotient in the rats were obtained from Arco system (Chiba, Japan). The system consisted of six acrylic metabolic chambers, a mass spectrometer (model WSMR-1400), a gas sampler (model WGSS-1000), and a switching controller (model WMSC-2000). Each metabolic chamber had a 210 cm2 floor and was 11.5 cm in height. Room air was pumped through the chambers at a rate 1.8 L/min. Expired air was dried in a cotton-thin column and then directed to a mass spectrometer. Air from each chamber was sampled for 1 min. Therefore, air from each chamber was measured every 7 min, and data were recorded in a spreadsheet. The respiratory quotient (RQ) and energy production rate were calculated by the software using the following formulas: RQ = VCO2/VO2 and energy production rate = (2.96*RQ + 2.49)*VO2, where VO2 is the oxygen consumption, and VCO2 is the carbon dioxide exhaustion. Additionally, 24 h urine samples were collected, and oxygen production from protein metabolism (VO2-P) was calculated from the urinary nitrogen concentration (UN). The UN was determined using an assay kit (Urinary nitrogen test from Wako Pure Chemicals, Tokyo, Japan), and VO2-P was calculated using the following formula: VO2-P = UN*5.923. Thus, the energy metabolism was compared between the two groups using the values of non-protein oxygen consumption and non-protein energy expenditure. After 1 week or 3 weeks of consuming the semi-synthetic diets, each animal was placed into a metabolic chamber for 24 h respiratory gas analysis. During the analysis, the animals were pair-fed and had free access to water. The sampled gases were analyzed in a separate room to avoid stressing the rats.
Analysis of mRNA expression
Total RNA was extracted from 0.1 g of WAT using an RNeasy Lipid Tissue Mini Kit (Qiagen, Tokyo, Japan). A TaqMan Universal PCR Master Mix (Applied Biosystems, Tokyo, Japan) and Assay-on-Demand Gene Expression Products (Rn00569117_m1 for fatty acid synthase (FAS), Rn00580702_m1 for carnitine palmitoyltransferase (CPT), Rn00566193_m1 for peroxisome proliferator-activated receptor (PPAR)-α, Rn00565707_m1 for PPAR-δ, Rn00440945_m1 for PPAR-γ, and Hs99999901_s1 for 18S RNA, Applied Biosystems) were used for quantitative real-time RT-PCR analysis of FAS, CPT, PPARα, PPARδ, PPARγ, and 18S RNA expression in the liver. Amplification was performed with a real-time PCR system (ABI Prism 7000 Sequence Detection System; Applied Biosystems). The results were quantified with a comparative method and expressed as a relative value after normalization to 18S RNA expression.
All of the values are expressed as the mean ± standard error. The significance of the differences between the means of the two groups was determined by Student’s t-test. Differences were considered to be significant at p < 0.05.
Results and discussion
Effects of the resveratrol diet on growth parameters and lipid parameters in OLETF rats
The effect of resveratrol on growth and lipid parameters in OLETF rats
Initial body weight (g)
132 ± 3
132 ± 3
Final body weight (g)
294 ± 3
299 ± 3
Food intake (g)
498 ± 3
499 ± 3
Serum cholesterol (mg/dL)
135 ± 4
122 ± 5
Serum triglyceride (mg/dL)
76.3 ± 5.0
96.5 ± 10.6
Hepatic cholesterol (mg/g liver)
3.34 ± 0.11
3.35 ± 0.13
Hepatic triglyceride (mg/g liver)
14.8 ± 1.4
16.4 ± 1.4
Effects of the resveratrol diet on nutrients oxidation in OLETF rats
Effects of the resveratrol diet on the serum glucose level and hepatic glycogen content in OLETF rats
A growing amount of evidence has demonstrated that some strategies can make the body recruit fat rather than glycogen to meet energy demands . A previous study demonstrated that AMPK activity and fuel selection in muscle in response to exercise can be manipulated by diet . Another study also reported that AMPK activation in the liver leads to the stimulation of fatty acid oxidation and inhibition of lipogenesis, glucose production and protein synthesis . Because dietary resveratrol has the potential to activate AMPK , it is possible that resveratrol induced sparing actions for carbohydrates and protein (which contribute to the anti-obesity effect) in OLETF rats.
Effects of the resveratrol diet on adipose mRNA levels in OLETF rats
In conclusion, our study showed that resveratrol has the potential to suppress body fat accumulation in non-high-fat diet-fed obese OLETF rats. The effect was, at least in part, attributable to the enhancement of fat oxidation and sparing actions for carbohydrates and protein in resveratrol-fed OLETF rats. Further studies are necessary to evaluate the effect of dietary resveratrol on nutrient metabolism in several animal models and determine the lowest effective concentration.
Fatty acid synthase
Otsuka Long-Evans Tokushima fatty
Peroxisome proliferator-activated receptor
White adipose tissue.
This work was supported by a research grant from the Japanese Ministry of Education, Culture, Sports, Science and Technology.
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