Aim. We investigated different bioactive compounds including epigallocatechin gallate (EGCG), anthocyanidin, resveratrol, phloretin, spermidine, butyrate, and β-hydroxybutyrate with regard to their effect on SIRT3 via NRF2 and modulation of the proinflammatory senescence-associated secretory phenotype (SASP) in senescence induced 3T3-L1 preadipocytes. Methods. For induction of senescence, 3T3-L1 preadipocytes were incubated with bromodeoxyuridine (BrdU) for 8 days. Cell cycle inhibition was observed, and β-galactosidase activity was measured. After BrdU treatment, cells were treated with different bioactive compounds in various concentrations for 96 h. ELISA was used for determining proinflammatory cytokine IL6 in SASP cells. Results. CDKN1a increased significantly after BrdU incubation compared to untreated control ( p < 0.01 ). All secondary plant ingredients used for treatment, but not anthocyanidin 50 μM, decrease CDKN1a expression ( p < 0.05 ), whereas most endogenous substances did not attenuate CDKN1a. IL6 secretion positively correlated with CDKN1a ( p < 0.01 ), whereas EGCG could diminish both, IL6 and CDKN1a with the strongest effect ( p < 0.01 ). Although NRF2 positively correlated with SIRT3 activation ( p < 0.05 ), only resveratrol ( p < 0.01 ) and anthocyanidin ( p < 0.05 ) could activate NRF2 significantly. Solely anthocyanidin 50 μM ( p < 0.05 ) and 100 μM ( p < 0.01 ) and EGCG 50 μM ( p < 0.01 ) could increase SIRT3 expression. Activation of SIRT3 with EGCG correlated with lowered IL6 secretion significantly ( p < 0.05 ) but not with anthocyanidin. Conclusion. Accumulation of senescent cells in adipose tissue plays an important role in obesity and age-related diseases. SIRT3, located in the mitochondria, can regulate ROS via different pathways. Thus, targeting SIRT3 activating compounds such as EGCG may delay senescence of cells and senescence induced inflammatory processes.

Periodic fasting (PF) is an increasingly popular approach that assists in the management of metabolic and inflammatory diseases as well as in preventing mechanisms involved in aging. However, little is known about the effects of fasting on gut microbiota and its impact on the epigenetic regulation of metabolically relevant enzymes, especially sirtuins (SIRTs). We analyzed the effect of periodic fasting on the human gut microbiota, SIRTs expression, and mitochondrial content in 51 males and females. The participants fasted under supervision for five consecutive days following the Buchinger fasting guidelines. Ketogenesis, selected mRNAs, miRNAs, mitochondrial (mt) DNA, and gut composition were analyzed before and after PF. PF triggered a significant switch in metabolism, as indicated by the increase in ß-hydroxybutyrate (BHB) and pyruvate dehydrogenase kinase isoform 4 (PDK4) expression in the capillary blood. MtDNA, SIRT1, SIRT3, and miRlet7b-5p expression in blood cells were elevated, whereas SIRT6 and miR125b-5p were not affected. Following fasting, gut microbiota diversity increased, and a statistically significant correlation between SIRT1 gene expression and the abundance of Prevotella and Lactobacillus was detected. The abundance of longevity related Christensenella species increased after fasting and inversely correlated with age as well as body mass index (BMI). Thus, this represents the first study that showing that fasting not only changes the composition of the gut microbiota, making it more diverse, but also affects SIRT expression in humans.

Healthy mitochondria and their epigenetic control are essential to maintaining health, extending life expectancy, and improving cardiovascular performance. Strategies to maintain functional mitochondria during aging include training; cardiovascular exercise has been suggested as the best method, but strength training has also been identified as essential to health and healthy aging. We therefore investigated the effects of concurrent exercise training and dietary habits on epigenetic mechanisms involved in mitochondrial (mt) functions and biogenesis. We analyzed epigenetic biomarkers that directly target the key regulator of mitochondrial biogenesis, PGC-1α, and mtDNA content. Thirty-six healthy, sedentary participants completed a 12-week concurrent training program. Before and after the intervention, dried blood spot samples and data on eating habits, lifestyle, and body composition were collected. MiR-23a, miR-30e expression, and mtDNA content were analyzed using real-time quantitative polymerase chain reaction (qPCR) analysis. PGC-1α methylation was analyzed using bisulfite pyrosequencing. MiR-23a, miR-30e expression, and PGC-1α methylation decreased after the intervention (p < 0.05). PGC-1α methylation increased with the consumption of red and processed meat, and mtDNA content increased with the ingestion of cruciferous vegetables (p < 0.05). Our results indicate that concurrent training could improve mitochondrial biogenesis and functions by altering the epigenetic regulation. These alterations can also be detected outside of the skeletal muscle and could potentially affect athletic performance.

Background: Fasting and fasting mimetics - bioactive compounds mimicking fasting effects, are of growing interest as potential means to slow down the aging process and increase health span. Sirtuins are known as enzymes that interfere with mitochondrial energy metabolism and molecular pathways involved in longevity. Although their activation is determined as a response to stress i.e. caloric restriction. Sirtuin activating nutraceuticals are believed to mimic the effects of nutrient deprivation, thus activating signaling pathways correlated to an improved health span. In this study, we compare 5 days periodic buchinger fasting intervention with 3 months shot supplementation, a drink formula, containing secondary plant ingredients considered to activate sirtuins.Methods: We analyzed pathways in response to fasting and a sirtuins activating drink. Genetic and epigenetic biomarkers including telomere length, LINE1 methylation, and a set of mRNAs and miRNAs were assessed using qPCR analysis. Gut composition and metabolites were compared using Illumnia sequencing and mass spectrometry.Results: Fasting, but also the fasting mimetic could increase expression of FoxO1, SIRT1, and MLH1 mRNA, all genes discussed in aspects of longevity. A positive correlation between telomere length and both SIRT1, and SIRT6 was observed. Furthermore, a significant change in the gut composition was measured. Actinobacteria increased in the supplementation group, whereas after buchinger fasting a rise in the distribution of Proteobacteria could be observed. Firmicutes/Bacteroidetes ratio decreased and correlated with the body mass index (BMI).Conclusions: Our results confirm the effects of fasting on longevity associated mechanisms but also suggest that SIRTFOOD shot intervention addresses some of these effects.

Background Regular, especially sustained exercise plays an important role in the prevention and treatment of multiple chronic diseases. Some of the underlying molecular and cellular mechanisms behind the adaptive response to physical activity are still unclear, but recent findings suggest a possible role of epigenetic mechanisms, especially miRNAs, in the progression and management of exercise-related changes. Due to the combination of the analysis of epigenetic biomarkers (miRNAs), the intake of food and supplements, and genetic dispositions, a “fitness score” was evaluated to assess the individual response to nutrition, exercise, and metabolic influence. Methods In response to a 12-week sports intervention, we analyzed genetic and epigenetic biomarkers in capillary blood from 61 sedentary, healthy participants (66.1% females, 33.9% males, mean age 33 years), including Line-1 methylation, three SNPs, and ten miRNAs using HRM and qPCR analysis. These biomarkers were also analyzed in a healthy, age- and sex-matched control group (n, 20) without intervention. Food frequency intake, including dietary supplement intake, and general health questionnaires were surveyed under the supervision of trained staff. Results Exercise training decreased the expression of miR-20a-5p, −22-5p, and −505-3p (p < 0.02) and improved the “fitness score,” which estimates eight different lifestyle factors to assess, nutrition, inflammation, cardiovascular fitness, injury risk, regeneration, muscle and hydration status, as well as stress level. In addition, we were able to determine correlations between individual miRNAs, miR-20a-5p, −22-5p, and −101-3p (p < 0.04), and the genetic predisposition for endurance and/or strength and obesity risk ( ACE, ACTN3 , and FTO ), as well as between miRNAs and the body composition (p < 0.05). MiR-19b-3p and −101-3p correlated with the intake of B vitamins. Further, miR-19b-3p correlated with magnesium and miR-378a-3p with iron intake (p < 0.05). Conclusions In summary, our results indicate that a combined analysis of several biomarkers (miRNAs) can provide information about an individual’s training adaptions/fitness, body composition, nutritional needs, and possible recovery. In contrast to most studies using muscle biopsies, we were able to show that these biomarkers can also be measured using a minimally invasive method.

Introduction: Topical investigations have demonstrated that oxidative stress and inflammation play key roles in biological aging and determine incidence and course of age-related diseases. Lifestyle and environmental factors hugely impact epigenetic regulation and DNA stability with telomere attrition and epigenetic instability providing a potential record of the cumulative burden of endogenous and exogenous oxidative noxae. Certain physiologically active plant components exhibit antioxidative activities affecting epigenetic regulation of inflammation response and DNA repair.Methods: Against this background, the present study investigated green tea polyphenol epigallocatechin gallate (EGCG) in the context of telomere regulation in Caco-2 colorectal adenocarcinoma cells vs. ES-1 primary skin fibroblasts. Cell lines were treated with 20 and 200 µM EGCG for 36, 72 and 144 hours, respectively. Telomerase activity, relative telomere length as well as methylation status of hTERT and c-Myc from different culture conditions were assessed. Malondialdehyde (MDA) served as a surrogate marker of potential pro-oxidative effects of EGCG in a physiologically relevant tissue model.Results: EGCG incubation was associated with telomere shortening and decreased telomerase activity in Caco-2 cells, and relatively longer telomeres along with increased methylation of six 5'—C—phosphate—G—3' (CpG) sites in the promoter region of human Telomerase Reverse Transcriptase (hTERT) in fibroblasts. At low concentrations, EGCG significantly decreased oxidative damage to lipids in Caco-2 cells and attenuated H2O2 induced oxidation at higher concentrations.Conclusion: These results suggest differential EGCG-mediated telomeric modulation in cancer vs. primary cells and a specific antioxidant activity of EGCG against oxidative damage to lipids in abnormal cells.Keywords: Caco-2, epigallocatechin gallate, telomeres, hTERT, DNA methylation, telomerase, oxidative stress, malondialdehyde

Background: Sirtuins attract high attention considering their properties to reverse molecular hallmarks of aging and age-related disorders. Many secondary plant ingredients (SPI) are known for their sirtuin-activating activities as well as epigenetic regulation of telomers, autophagy, senolysis, DNA repair but also improvement of gut microbiota. Furthermore, prebiotics enhanced butyrate was shown to interact with SIRT pathways. This study investigated effects of a drink containing a mix of different SPIs in combination with galactooligosaccharides (GOS) and their effect on SIRT activation, markers of aging relevant mechanisms and gut microbiota composition in correlation with subjective wellbeing and skin structure appearance. Methods: We analyzed gene expression, mtDNA amount, and microbial composition in response to a sirtuin-activating drink in humans compared to a control group consuming a placebo. Food frequency, beauty, and general health questionnaires were asked, and a set of mRNAs and miRNAs were assessed using qPCR analysis. The gut composition was analyzed using Illumnia sequencing.Results: SPI increased SIRT1, SIRT3 and modulated cell cycle relevant miR16 and senescence regulating miR34 expression. Additionally, mtDNA amount was higher in the group consuming the active supplement indicating an improved mitochondrial activity. The combined effect of SPI and GOS lead to an increase of Actinobacteria, especially Bifidobacterium, but also Veillonellaceae which was not observed in the control group. Significant correlations between SIRT3 expression and the gut microbiota Bifidobacterium and Veillonellaceae were observed. Additionally, statistical analysis of subjects self-reporting indicated beneficial effects regarding beauty and wellbeing.Conclusion: Our results show that the combination of sirtuins inducing SPI and prebiotic GOS influences molecular pathways counteracting aging, senescence, inflammation, and enhanced groups of gut microbiota which are known to improve the innate and adaptive immune system. Keywords: secondary plant ingredients, prebiotic, Sirtuins, subjective wellbeing, Bifidobacterium

Background: Regular, especially sustained exercise training plays an important role in the prevention and treatment of multiple chronic diseases. Some of the underlying molecular and cellular mechanisms behind the adaptive response to physical activity are still unclear, but recent findings suggest a possible role of epigenetic mechanisms, especially miRNAs, in the progression and management of exercise related changes. Due to the combination of the analysis of epigenetic biomarkers (miRNAs), the intake of food and supplements, and genetic dispositions, a “fitness sore” was evaluated to assess the individual response to nutrition, metabolism and exercise.Methods: In response to a 12-week sport intervention we analyzed genetic and epigenetic biomarkers in capillary blood, including Line-1 methylation, three SNPs and ten miRNAs using HRM and qPCR analysis. These biomarkers were also analyzed in a control group without intervention. Food frequency intake, including dietary supplement intake, and general health questionnaires were surveyed under the supervision of trained staff.Results: Exercise training decreased the expression of miR-20a, -22 and -505 (p < 0.02) and improved the “fitness score”, which estimates eight different lifestyle factors to assess, nutrition, inflammation, cardiovascular fitness, injury risk, regeneration, muscle- and hydration status as well as stress level. In addition, we were able to determine correlations between individual miRNAs, miR-20a, -22 and -101 (p < 0.04), and the genetic predisposition for endurance and / or strength and for obesity risk (ACE, ACTN3 and FTO), as well as between miRNAs and the body composition (p < 0.05). And we identified two miRNAs, miR-19b and -378a (p < 0.05), which could potentially provide information about the micronutrient / vitamin requirements of an athlete.Conclusions: Due to the detailed knowledge of individual regulatory mechanisms in the metabolism of sport intervention and / or nutritional behavior, this knowledge and our results can be used for personalized interventions and in the context of the new field of precision nutrition and precision training.