Mech Ageing Dev. 2005 Jun 4.
Calorie restriction and SIR2 genes-Towards a mechanism.
Guarente L.
Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.

Calorie restriction is the first and most compelling example of life extension in mammals. Much speculation about how CR works has focused on ideas of what causes aging. Since these causes themselves are much disputed, I have instead focused my thinking on lessons from simple model organisms, which have emerged from recent genetic studies. These findings can now be integrated with numerous, elegant studies on CR over the decades, which provide a treasure trove of information about physiological changes that are elicited by this regimen. In this paper, I present data showing that the SIR2 gene is a strong candidate to regulate CR in the simple model organisms, such as yeast and Drosophila. I then summarize what is known about the mammalian Sirt1 as it relates to physiological changes during CR, and discuss how this mechanism may impact on life span, as well as diseases of aging.

Mech Ageing Dev. 2005 May 10.
Membrane alteration as a basis of aging and the protective effects of calorie restriction.
Yu BP.
Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.

As has been experimentally determined, oxidative modification to biological systems can be extensive, although the identification and stochiometric relation of the reactive species that cause these alterations have not been fully elucidated. In this review, arguments are presented to support the notion that the combined effects of membrane lipid peroxidation and its by-products, reactive aldehydes are likely responsible for membrane-associated functional declines during aging. As evidence for a systemic response to overall oxidative stress, the molecular inflammation hypothesis of aging is discussed by considering that the activation of inflammatory genes act as a bridge linking normal aging to pathological processes.

Biogerontology. 2005;6(1):27-37.
The effect of age and calorie restriction on HIF-1-responsive genes in aged liver.
Kang MJ, Kim HJ, Kim HK, Lee JY, Kim DH, Jung KJ, Kim KW, Baik HS, Yoo MA, Yu BP, Chung HY.
College of Pharmacy, Aging Tissue Bank, Pusan National University, Busan, 609-735, Korea.

Hypoxia inducible factor-1 (HIF-1) regulates transactivation of several genes in response to hypoxia condition. We explore hepatic HIF-1 responsive gene regulation during aging and the age-related changes of the HIF-1 related gene activation in young and old rats. Results indicate that the aging process induces the activation of HIF-1alpha, which is accompanied by increased HIF-1 DNA binding. This increased binding activity is accompanied by the increase of HIF-1-dependent genes, heme oxygenase-1 (HO-1), vascular endothelial growth factor (VEGF), erythropoietin (EPO), and inducible nitric oxide synthase (iNOS), which all showed remarkable up-regulation during aging process. In contrast, the increased HIF-1 related gene expression was effectively blunted by the anti-oxidative action of calorie restriction in aged rat liver. We propose that age-related HIF-1 binding activity may well be influenced by the increased pro-oxidative conditions of aged animals, which up-regulate HIF-1-dependent gene expression.

Nutrition. 2005 Mar;21(3):378-88.
Caloric restriction modulates insulin receptor signaling in liver and skeletal muscle of rat.
Zhu M, de Cabo R, Anson RM, Ingram DK, Lane MA.
Laboratory of Experimental Gerontology, Gerontology Research Center, Intramural Research Program, National Institute on Aging, Baltimore, Maryland, USA.

OBJECTIVE: We investigated how the insulin/insulin-like growth factor-1 signaling pathway is involved in the robust antiaging effects produced by caloric restriction. METHODS: We subjected male rats to feeding ad libitum or calorie restriction, i.e., 60% of the ad libitum amount, for 2 and 25 mo and then assessed the effects of calorie restriction on insulin receptor (IR) signaling in liver and skeletal muscle. RESULTS: The results indicated that aging was accompanied by a significant decrease in IR tyrosine phosphorylation after insulin stimulation in live and skeletal muscle, which was associated with a significant increase in the activity of protein tyrosine phosphatase-1B. However, these age-related alterations were attenuated by long-term calorie restriction. Expression profile of mRNA showed an increased expression of mRNAs for IR and insulin-like growth factor-1 receptor in both tissues of calorie-restricted rats, but increased expression of IR mRNA was dissociated with the IR gene product in rats maintained on long-term calorie-restricted diet. CONCLUSION: IR signaling may play an important role in aging and its retardation by calorie restriction, and normal function of IR in liver and skeletal muscle is required for healthy aging and extending lifespan in mammals.

Ageing Res Rev. 2005 Jan;4(1):55-65. Epub 2004 Dec 8.
Lessons learned from gene expression profile studies of aging and caloric restriction.
Park SK, Prolla TA.
Department of Genetics and Medical Genetics, University of Wisconsin, 5302B Genetics building, 445 Henry Mall, Madison, WI 53706, USA.

To examine molecular events associated with aging and its retardation by caloric restriction (CR), we have employed high-density oligonucleotide microarrays to define transcriptional patterns in mouse tissues, including skeletal muscle, brain, heart, and adipose. Aging results in a differential gene expression pattern specific to each tissue, and most alterations can be completely or partially prevented by CR. Transcriptional patterns of tissues from calorie-restricted animals suggest that CR retards the aging process by reducing endogenous damage and by inducing metabolic shifts associated with specific transcriptional profiles. These studies demonstrate that DNA microarrays can be used in aging research to generate panels of hundreds of transcriptional biomarkers, providing a new tool to measure biological age on a tissue-specific basis and to evaluate interventions designed to mimic the effects of CR.

Endocrinology. 2005 Apr;146(4):2098-106.
Moderate caloric restriction, but not physiological hyperleptinemia per se, enhances mitochondrial oxidative capacity in rat liver and skeletal muscle--tissue-specific impact on tissue triglyceride content and AKT activation.
Barazzoni R, Zanetti M, Bosutti A, Biolo G, Vitali-Serdoz L, Stebel M, Guarnieri G.
Clinica Medica, University of Trieste, Ospedale Cattinara, Strada di Fiume 447, 34100 Trieste, Italy.

The study aimed at determining, in lean tissues from nonobese rats, whether physiological hyperleptinemia with leptin-induced reduced caloric intake and/or calorie restriction (CR) per se: 1) enhance mitochondrial-energy metabolism gene transcript levels and oxidative capacity; and 2) reduce triglyceride content. Liver and skeletal muscles were collected from 6-month-old Fischer 344 rats after 1-wk leptin sc infusion (0.4 mg/kg . d: leptin + approximately 3-fold leptinemia vs. ad libitum-fed control) or moderate CR (-26% of those fed ad libitum) in pair-fed animals (CR). After 1 wk: 1) leptin and CR comparably enhanced transcriptional expression of mixed muscle mitochondrial genes (P < 0.05 vs. control); 2) CR independently increased (P < 0.05 vs. leptin-control) hepatic mitochondrial-lipooxidative gene expression and oxidative capacity; 3) hepatic but not muscle mitochondrial effects of CR were associated (P < 0.01) with increased activated insulin signaling at AKT level (P < 0.05 vs. leptin-control); 4) liver and muscle triglyceride content were comparable in all groups. In additional experiments, assessing time course of posttranscriptional CR effects, 3-wk superimposable CR (P < 0.05): 1) increased both liver and muscle mitochondrial oxidative capacity; and 2) selectively reduced muscle triglyceride content. Thus, in nonobese adult rat: 1) moderate CR induces early increments of mitochondrial-lipooxidative gene expression and time-dependent increments of oxidative capacity in liver and mixed muscle; 2) sustained moderate CR alters tissue lipid distribution reducing muscle but not liver triglycerides; 3) mitochondrial-lipid metabolism changes are tissue-specifically associated with hepatic AKT activation; 4) short-term physiological hyperleptinemia has no independent stimulatory effects on muscle and liver mitochondrial-lipooxidative gene expression. Increased lean tissue oxidative capacity could favor substrate oxidation over storage during reduced nutrient availability.

FEBS Lett 1999 Sep 17;458(2):231-5.
Calorie restriction increases Fas/Fas-ligand _expression and apoptosis in murine splenic lymphocytes.
Reddy Avula CP; Muthukumar A; Fernandes G.
Division of Clinical Immunology, Department of Medicine, The University of Texas Health Science Center at San Antonio 78229-3900, USA.

One-month-old male ICR mice were fed a nutritionally adequate, semipurified diet, either ad libitum (AL) or calorie restricted (CR) (40% less food) for 6 months and were killed to obtain spleens. Flow cytometric analysis revealed increased proportions of both CD4+ and CD8+ T cells in CR-fed mice compared to AL-fed mice. The T cell subsets of CR-fed mice were also found to have higher levels of plasma membrane Fas receptor _expression. Similarly, Fas-ligand (Fas-L) _expression was higher in anti-CD3-stimulated CD4+ and CD8+ T cells. CR-fed mice also had increased numbers of annexin V-positive CD4+ and CD8+ T cells in stimulated splenic lymphocytes suggesting an increased potential for apoptosis. Fas and Fas-L gene _expression in splenic lymphocytes, which correlated closely with the observed increased rate of apoptosis, was significantly increased in CR-fed mice compared to AL-fed mice. In conclusion, these results indicate that CR increases the _expression of Fas and Fas-L which may contribute to the known beneficial effects of CR such as prolongation of life span by activating chronic physiologically mediated apoptosis.

Toxicol Sci 1999 Dec;52(2 Suppl):56-60.
Calorie restriction in nonhuman primates: mechanisms of reduced morbidity and mortality.
Hansen BC; Bodkin NL; Ortmeyer HK.
Obesity and Diabetes Research Center, University of Maryland, Baltimore 21201, USA.

Long term chronic calorie restriction (CR) of adult nonhuman primates significantly reduces morbidity and increases median age of death. The present review is focused upon an ongoing study of sustained adult-onset calorie restriction, which has been underway for 15 years. Monkeys, initially calorie restricted at about 10 years of age, are now approximately 25 years old. The median life span of these restricted monkeys is increasing, now exceeding that of ad libitum (AL)-fed monkeys. In our laboratory, maximum life span for AL-fed monkeys appears to be about 40 years. Thus, whether CR can also increase maximal life span, as it does in rodents, cannot be determined for at least another 15 years. The earliest detectable positive benefit on morbidity in these monkeys was previously reported as the prevention of obesity. Current evidence, as reviewed here, suggests that much obesity-associated morbidity is also mitigated by sustained calorie restraint in nonhuman primates. Furthermore, probably because of the prevention of obesity, diabetes has also been prevented. Recent findings include the identification of extraordinary changes in the glycogen synthesis pathway, and on the phosphorylation of glycogen synthase in response to insulin. This calorie restriction-induced prevention of morbidity does not require excessive leanness, but is clearly present when body fat is within the normal range of 10 to 22%, and this is likely to be true in humans as well.

Eur J Cell Biol 1999 Aug;78(8):573-9.
Fasting-induced apoptosis in rat liver is blocked by cycloheximide.
Tessitore L; Tomasi C; Greco M.
Dipartimento di Scienze Mediche, Universita del Piemonte Orientale, Novara, Italy.

The effect of cycloheximide (CH) on the fasting-induced changes of rat liver cell and protein turnover has been investigated. Late starvation phase (3-4-day-fasting period) was characterised by a decrease in liver weight and protein and DNA content. The loss of DNA was not related to liver cell necrosis but due not only to depression of cell proliferation as shown by the drop in the labelling index but also induction of apoptosis. This type of apoptosis was documented by the increase in the apoptotic index (cells labelled by TUNEL) and transglutaminase activity as well as by DNA fragmentation. The liver cells of fasted rats appeared smaller as shown by the higher cell density and DNA/protein ratio than in controls. Females were more resistant to fasting-induced apoptosis than males. A single dose of CH, a drug primary known as inhibitor of protein synthesis, induced or enhanced apoptosis in fed and 2-days fasted male rats, respectively, without any sign of cell necrosis. On the contrary, the administration of repeated doses of CH blocked apoptosis induced by fasting. CH "froze" protein and DNA content as well as apoptotic process at the level of 2 days-fasted rats. While fasting-induced liver protein loss resulted from a marked reduction in protein synthesis with a slight decrease in degradation, repeated treatment with CH virtually blocked protein loss by abolishing protein catabolism. These data suggest a direct relationship between the catabolic side of protein turnover and the apoptotic process.

J Gerontol A Biol Sci Med Sci 1998 May;53(3):B168-72.
Diet restriction increases apoptosis in the gut of aging rats.
Holt PR; Moss SF; Heydari AR; Richardson A.
Department of Medicine, St. Luke's/Roosevelt Hospital Center, New York, USA.

Previous studies have shown that epithelial cell production rates are increased throughout the gastrointestinal tract in aging rats. We tested the hypothesis that alteration in cell death (apoptosis) might be involved. Fischer 344 rats aged 4-5 months and 24-25 months fed ad libitum (AL) or calorie restricted (CR) to 60% of the AL intake were studied. Epithelial cell apoptosis was determined by a terminal deoxyuridine nucleotidyl nick end labeling (TUNEL) technique validated in our laboratory, and the _expression of four members of the Bcl-2 family was evaluated by Western blotting in the small intestine and colon. The apoptotic index was low in young and aging AL and young CR rats. However, CR in aging rats was associated with a significantly higher apoptotic index in the jejunum and colon. The _expression of the Bcl-2 family of genes was unchanged. Enhanced apoptosis in CR may protect the gastrointestinal tract from accumulation of DNA-altered cells during the aging process.

J Nutr 1995 Feb;125(2):195-201.
Activities of antioxidant enzymes in various tissues of male Fischer 344 rats are altered by food restriction.
Xia E; Rao G; Van Remmen H; Heydari AR; Richardson A.
Geriatric Research, Education and Clinical Center, Audie L. Murphy Memorial Veterans Hospital, San Antonio, TX.

ABSTRACT: The objective of this study was to determine how food restriction (40% restriction of food intake) altered the age-related changes in the activities of Cu,Zn superoxide dismutase, catalase and glutathione peroxidase in liver, brain cortex, heart, kidney and intestinal mucosa obtained from 6-, 16- and 26-mo-old male Fischer 344 rats. Food restriction increased the activity of one or more of the antioxidant enzymes in the liver, brain cortex, heart and kidney of the rats. However, the magnitude of the effect and the antioxidant enzyme(s) affected by food restriction varied from tissue to tissue, and food restriction had no significant effect on the activities of these enzymes in intestinal mucosa. Interestingly, the four tissues in which food restriction increased the activity of one or more of the antioxidant enzymes showed reduced lipid peroxidation as measured by thiobarbituric acid-reactive material. These data suggest that food restriction might enhance the survival of rodents by altering the levels of the antioxidant enzymes and hence reducing free radical damage.