Resveratrol improves health and survival of mice on a high-calorie diet

Abstract

Resveratrol (3,5,4'-trihydroxystilbene) extends the lifespan of diverse species including Saccharomyces cerevisiae, Caenorhabditis elegans and Drosophila melanogaster. In these organisms, lifespan extension is dependent on Sir2, a conserved deacetylase proposed to underlie the beneficial effects of caloric restriction. Here we show that resveratrol shifts the physiology of middle-aged mice on a high-calorie diet towards that of mice on a standard diet and significantly increases their survival. Resveratrol produces changes associated with longer lifespan, including increased insulin sensitivity, reduced insulin-like growth factor-1 (IGF-I) levels, increased AMP-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha) activity, increased mitochondrial number, and improved motor function. Parametric analysis of gene set enrichment revealed that resveratrol opposed the effects of the high-calorie diet in 144 out of 153 significantly altered pathways. These data show that improving general health in mammals using small molecules is an attainable goal, and point to new approaches for treating obesity-related disorders and diseases of ageing.

Figure 1. Resveratrol increases survival and improves rotarod performance

a, Body weights of mice fed a standard diet (SD), high-calorie diet (HC), or high-calorie diet plus resveratrol (HCR). b, Kaplan–Meier survival curves. Hazard ratio for HCR is 0.69 (χ² = 5.39, P = 0.020) versus HC, and 1.03 (χ² = 0.022, P = 0.88) versus SD. The hazard ratio for HC versus SD is 1.43 (χ² = 5.75, P = 0.016). c, Time to fall from an accelerating rotarod was measured every 3 months for all survivors from a pre-designated subset of each group; n = 15 (SD), 6 (HC) and 9 (HCR). Asterisk, P < 0.05 versus HC; hash, P < 0.05 versus SD. Error bars indicate s.e.m.

Figure 2. Resveratrol improves insulin sensitivity and activates AMPK

a–d, Plasma levels of glucose (a, b) and insulin (c, d) were measured after a 2 g kg⁻¹ oral glucose dose. Areas under the curves (AUC) were significantly reduced by resveratrol treatment. e, Activation of AMPK by resveratrol in CHO cells. In the presence of resveratrol or 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) as a positive control, phosphorylation of AMPK and its downstream target, acetyl-coA carboxylase (ACC), are increased. f, AMPK activity in liver. Phosphorylation of AMPK (f), acetyl-coA carboxylase (Supplementary Fig. 3e) and decreased expression of fatty acid synthase (Supplementary Fig. 3f) are indicative of enhanced AMPK activity. Asterisk, P < 0.05 versus HC; hash, P <0.05 versus SD. n = 5 for all groups. Error bars indicate s.e.m.

Figure 3. Resveratrol improves liver histology, increases mitochondrial number and decreases acetylation of PGC-1α

a–c, Resveratrol prevents the development of fatty liver, as assessed by organ size (a), overall pathology (b) and decreased fat accumulation as measured by oil red O staining (c). AU, arbitrary units. d, Pathology of heart sections. Additional histology of liver, heart and aorta is shown in Supplementary Fig. 4. e, f, Transmission electron microscopy of liver sections (e) and mitochondrial counts (f). g, h, Mitochondrial number in HeLa cells treated with serum from ad libitum-fed (AL) or calorically restricted (CR) rats, or resveratrol, and stained with Mitotracker green FM. i, j, Resveratrol reduces the acetylation of PGC-1α, a known SIRT1 target and regulator of mitochondrial biogenesis, in vivo. PGC-1α was immunoprecipitated from liver extracts then blotted for acetyl lysine (i) and quantified (j). Asterisk, P < 0.05 versus HC; hash, P < 0.05 versus SD. n = 5 for b and d; n = 3 for f and j. Error bars indicate s.e.m.

Figure 4. Resveratrol shifts expression patterns of mice on a high-calorie diet towards those on a standard diet

a, b, The most highly significant upregulated (a) and downregulated (b) genes in livers of HC and HCR groups are shown. c, Parametric analysis of gene-set enrichment (PAGE) comparing every pathway significantly upregulated (red) or downregulated (blue) by either the HC diet or resveratrol (153 in total, with 144 showing opposing effects). d, Principal component analysis of PAGE data. The first principal component (PC1) is dominant, with 88.4% variability, and shows HCR to be more similar to SD than HC. e, Comparison of pathways significantly altered by resveratrol treatment and caloric restriction using data from the AGEMAP caloric restriction study. Pathways with significant differences between HC and HCR are indicated by an asterisk. Complete pathway listings are in Supplementary Fig. 7. Asterisk, P < 0.05 versus HC. n = 5 for SD and HC; n = 4 for HCR.