Different Antioxidant Profiles in Italian Centenarians: the Sardinian Peculiarity

Home , University of Perugia, University of Sassari

SHORT COMMUNICATION Different antioxidant profiles in Italian centenarians: the Sardinian peculiarity

MC Polidori1,6, E Mariani2,6, G Baggio3, L Deiana4, C Carru4, GM Pes4, R Cecchetti2, C Franceschi5, U Senin2 and P Mecocci2

1Institute of Biochemistry and Molecular Biology I, Heinrich-Heine Universita¨t Du¨sseldorf, Germany; 2Institute of Gerontology and Geriatrics, University of Perugia, Perugia, Italy; 3Internal Medicine Unit, Azienda Ospedale-Universita` di Padova, Italy; 4Department of Biomedical Sciences, Chair of Clinical Biochemistry, University of Sassari, Italy and 5Department of Experimental Pathology and C.I.G., University of Bologna, Italy

In this study, 153 Italian centenarians from four different geographical areas, including Modena (northern Italy), Ancona (central Italy), Perugia (central Italy) and Sardinia island (AKEA Project) were enrolled. Plasma levels of vitamin C, uric acid, vitamin A and vitamin E as well as the activities of superoxide dismutase and glutathione peroxidase were measured. Subjects were compared to a younger control population of the same areas, divided into three age groups: p60; 61–80 and 81–99 years. The main result of this study is that, whereas in Italian population elevated levels of plasma vitamins A and E seem to be important for longevity, this do not apply to centenarians from Sardinia, in which probably other factors play a more important role. European Journal of Clinical Nutrition (2007) 61, 922–924; doi:10.1038/sj.ejcn.1602596; published online 17 January 2007

Keywords: centenarians; antioxidants; oxidative stress; longevity; vitamins

Human longevity is a multifactorial condition with both population, in the present study we enrolled 153 centenar- genetic and environmental contribution. Among modifiable ians (113 females, 40 males, mean age 101.871.9 years) from protective factors allowing the achievement of advanced age four different geographical areas in Italy: 32 subjects from (Rattan, 2004), optimal antioxidant status of the organism is Modena (northern Italy), 20 subjects from Ancona (central considered to play an important role in counteracting the Italy), 30 subjects from Perugia (central Italy) and 71 subjects detrimental effects of age-associated free radical hyper- from Sardinia. Subjects were compared to a younger control production and oxidative stress (Meydani, 1999). population divided into three age groups: p60 years or In a relatively small sample of centenarians, which group A, n. 89 subjects (55 F, 34 M, mean age 53.675.0); represents a good model of human longevity, we previously 61–80 years or group B, n. 57 subjects (27 F, 30 M, mean age observed high plasma levels of vitamins A and E in the 70.675.9); 81–99 years or group C, n. 73 subjects (43 F, 30 M, presence of otherwise relatively poor plasma antioxidant mean age 91.576.1). Centenarians were defined as group D. status in comparison to younger subjects (Mecocci et al., All subjects, evaluated according to the study protocol 2000). To verify this finding in a larger centenarian (Deiana et al., 1999; Franceschi et al., 2000), were substan- tially healthy, free from major organ failure and well nourished. After informed consent, one 10-ml blood sample Correspondence: Professor P Mecocci, Institute of Gerontology and Geriatrics, University of Perugia, Policlinico Monteluce-Padiglione E, Via Brunamonti 51, (heparinized tube) was withdrawn and immediately centri- 06122 Perugia, Italy. fuged. Plasma was stored frozen at À801C until analysis. E-mail: [email protected] Vitamin C and uric acid were detected by high-perfor- 6 These authors contributed equally to this work mance liquid chromatography (HPLC) with electrochemical Contributors: MCP, EM and PM supervised the conduct of the study and drafted the manuscript. MCP and EM did all study assessment. LD and CC detection (Kutnink et al., 1987); vitamin A and vitamin E obtained the samples. RC and PM analysed study samples. GB, GMP, CF and were measured by HPLC with UV detection (Nierenberg and US were involved in study design and obtaining funding. EM and PM did Nann, 1992); superoxide dismutase (SOD) and glutathione sample size estimates and the statistical analyses. All contributors reviewed the peroxidase (GPx) activities were measured spectrophotome- manuscript. Received 25 May 2006; revised 6 November 2006; accepted 6 November trically, as previously described (Flohe´ and Gu¨nzler, 1984; 2006; published online 17 January 2007 L’Abbe´ and Fisher, 1990). Centenarians and antioxidants MC Polidori et al 923 Table 1 Plasma levels of vitamin C, uric acid, vitamin A and vitamin E and activities of SOD and GPx in healthy subjects of different age groups

Group A (p60 years) Group B (61–80 years) Group C (81–99 years) Group D (X100 years) n ¼ 89 n ¼ 57 n ¼ 73 n ¼ 153

Vitamin C (mM) 49.5714.5111*** 49.8715.8111*** 35.779.8*** 29.674.5 Uric acid (mM) 324.1788.0111*** 293.7779.911*** 243.2764.2 218.6757.4 Vitamin A (mM)2.670.3*** 2.770.2*** 2.470.3*** 3.571.8 Vitamin E (mM) 48.675.51 49.277.21 44.676.0*** 49.978.3 SOD (U/ml) 27.976.2* 28.676.1* 27.077.0 25.274.6 GPx (nmolNADPH/min/ml) 0.1270.01* 0.1270.02* 0.1170.02 0.1070.01

Abbreviations; GPx, glutathione peroxidase, SOD, super oxide dismutase. Data are expressed as mean7s.d. 1Po0.01, 11Po0.001, 111Po0.0001 vs group C. *Po0.01, **Po0.001, ***Po0.0001 vs group D.

Statistical analysis was performed with the program 80 ˚˚˚ Ancona 70 Statistical Package for the Social Sciences (SPSS version Modena 60 §§ 11.5, Chicago, IL, USA). ANOVA with Scheffe´ post hoc test 50 Perugia was used for comparisons between groups. All data are 40 30 *** *** ^ ^ Sardinia presented as mean7s.d. An overall P-value of at least 0.05 20 ^^ was required for significance. 10 ^^ As shown in Table 1, plasma levels of the two water-soluble 0 antioxidants vitamin C and uric acid were significantly SOD GPx higher in subjects of groups A and B compared to C and D, Vitamin C Uric Acid Vitamin A Vitamin E indicating an age-related decrease. These results are in agreement with our previous study (Mecocci et al., 2000). Figure 1 Plasma levels of vitamin C ((mM), uric acid (mM; x/10), vitamin A (mM; x Â 10) and vitamin E (mM) and activities of We also found a decrease in the activities of two potent superoxide dismutase (SOD) (U/ml) and glutathione peroxidase antioxidant enzymes, SOD and GPx, along with age. Finally, (GPx) (nmol NADPH/min/ml; x Â 100) in centenarians (female/male) in this large sample of centenarians, we confirmed the of Ancona (n ¼ 20; 17 F/3 M), Modena (n ¼ 32; 28 F/4 M), Perugia previously observed elevated plasma levels of the liposoluble (n ¼ 30; 24 F/6 M) and Sardinia (n ¼ 71; 44 F/27 M). Data are expressed as mean7s.d. ***Po0.001 vs Ancona, Modena and vitamins A and E compared to younger groups. However, this yy Perugia; 111Po0.001 vs Ancona and Modena; Po0.01 vs Ancona latter result did not apply to the centenarians from Sardinia, and Perugia; ^^Po0.01 ^Po0.05 vs Modena and Perugia. and, interestingly, their plasma vitamin A and E levels did not significantly differ from those of younger subjects of the same geographical area (data not shown). 2006), in addition to those linked to antioxidants and More in detail, Sardinian centenarians showed signifi- already known to play an important role in longevity. cantly higher levels of uric acid than centenarians from Ancona, Modena and Perugia, as well as significantly lower vitamin A and E levels. Their SOD and GPx activity was lower Acknowledgements compared to centenarians from Modena and Perugia, as observed also in centenarians from Ancona (Figure 1). LD was funded for the ‘AKEA Project’ by Regione Autonoma The main result of this study is the observation, in a large della Sardegna, MIUR Italy, the US National Institute on number of centenarians, of elevated levels of plasma Aging, Duke University no. 03-SC-NIH-1027 (USA), Max- vitamins A and E. This appears to be a characteristic of Planck Institute for Demographic Research (MPIDR). longevity in the Italian population from different geographic areas but does not seem to apply to the centenarians from Sardinia. So, it is conceivable that in Sardinia other factors, References in example genetic, rather than environmental factors play a major role in longevity compared to other Italian Caselli G, Pozzi L, Vaupel JW, Deiana L, Pes G, Carru C et al. (2006). populations. This aspect, for example, is supported by the Family clustering in Sardinian longevity: a genealogical approach. Exp Gerontol 41, 727–736. observation that in Sardinia the centenarians’ male: female Deiana L, Ferrucci L, Pes GM, Carru C, Delitala G, Ganau A et al. ratio is 1:2, being between 2 and 3 in Southern Italy, and (1999). AKEntAnnos. The Sardinian Study of extreme longevity. between 5 and 7 in Northern Italy (Passarino et al., 2002). Aging 11, 142–149. ´ This supports the idea of a peculiar background of Flohe L, Gu¨nzler WA (1984). Assays of glutathione peroxidase. Methods Enzymol 105, 114–121. Sardinian centenarians, probably associated to genetic and Franceschi C, Motta L, Valensin S, Rapisarda R, Franzone A, Berardelli demographic aspects still under evaluation (Caselli et al., M et al. (2000). Do men and women follow different trajectories to

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