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1.
Life (Basel) ; 14(3)2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38541714

ABSTRACT

The importance of diet and lifestyle in maintaining overall health has long been recognised. MicroRNAs (miRNAs) have emerged as key players in the intricate interplay between health and disease. This study, including 305 participants, examined the role of miRNAs from capillary blood as indicators of individual physiological characteristics, diet, and lifestyle influences. Key findings include specific miRNAs associated with inflammatory processes and dietary patterns. Notably, miR-155 was associated with subjects with metabolic diseases and upregulated in age. Additionally, the study revealed diet-related miRNA expressions: high consumption of vegetables, fruits, and whole grains correlated with increased levels of miR-let-7a and miR-328, both implicated in anti-inflammatory pathways, and decreased expression of pro-inflammatory miR-21. In the context of smoking, we found a significant decrease in miRNA-142, known for its downregulation in lung cancer. We observed a sex-biased expression of various miRNAs with significant upregulation of miR-151a in females and a higher expression of miR-155 in ageing females, representing a possible mechanism for the increased susceptibility to autoimmune diseases. In conclusion, the study underscores the significant influence of lifestyle, nutrition, and sex on miRNA profiles. Circulating miRNAs demonstrate significant potential as biomarkers in personalized medicine, highlighting their utility in tailoring healthcare to individual needs.

2.
Int J Mol Sci ; 22(5)2021 Feb 26.
Article in English | MEDLINE | ID: mdl-33652686

ABSTRACT

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.


Subject(s)
Clostridiales/growth & development , Fasting/blood , Gastrointestinal Microbiome , Gene Expression Regulation, Enzymologic , Sirtuins/biosynthesis , Adult , Aged , Female , Humans , Male , Middle Aged
3.
Oxid Med Cell Longev ; 2020: 4793125, 2020.
Article in English | MEDLINE | ID: mdl-33149809

ABSTRACT

AIM: We investigated different bioactive compounds including epigallocatechin gallate (EGCG), anthocyanidin, resveratrol, phloretin, spermidine, butyrate, and ß-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 ß-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.


Subject(s)
Catechin/analogs & derivatives , Cellular Senescence/drug effects , Sirtuin 3/metabolism , 3T3-L1 Cells , Animals , Anthocyanins/pharmacology , Bromodeoxyuridine/metabolism , Catechin/pharmacology , Cell Shape/drug effects , Cyclin-Dependent Kinase Inhibitor p21/genetics , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Gene Expression Regulation/drug effects , Genotype , Interleukin-6/metabolism , Mice , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , Polyphenols/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Resveratrol/pharmacology , Sirtuin 3/genetics , beta-Galactosidase/metabolism
4.
Oxid Med Cell Longev ; 2018: 3734250, 2018.
Article in English | MEDLINE | ID: mdl-29854080

ABSTRACT

Obesity- or diabetes-induced oxidative stress is discussed as a major risk factor for DNA damage. Vitamin E and many polyphenols exhibit antioxidative activities with consequences on epigenetic regulation of inflammation and DNA repair. The present study investigated the counteraction of oxidative stress by vitamin E in the colorectal cancer cell line Caco-2 under normal (1 g/l) and high (4.5 g/l) glucose cell culture condition. Malondialdehyde (MDA) as a surrogate marker of lipid peroxidation and reactive oxygen species (ROS) was analyzed. Gene expression and promoter methylation of the DNA repair gene MutL homolog 1 (MLH1) and the DNA methyltransferase 1 (DNMT1) as well as global methylation by LINE-1 were investigated. Results revealed a dose-dependent counteracting effect of vitamin E on H2O2-induced oxidative stress. Thereby, 10 µM vitamin E proved to be more efficient than did 50 µM in reducing MDA. Further, an induction of MLH1 and DNMT1 gene expression was noticed, accompanied by an increase in global methylation. Whether LINE-1 hypomethylation is a cause or effect of oxidative stress is still unclear. In conclusion, supplementation of exogenous antioxidants like vitamin E in vitro exhibits beneficial effects concerning oxidative stress as well as epigenetic regulation involved in DNA repair.


Subject(s)
DNA (Cytosine-5-)-Methyltransferase 1/genetics , DNA (Cytosine-5-)-Methyltransferases/metabolism , DNA Methylation/drug effects , MutL Protein Homolog 1/genetics , Oxidative Stress/drug effects , Vitamin E/pharmacology , Caco-2 Cells , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/genetics , Colorectal Neoplasms/metabolism , DNA (Cytosine-5-)-Methyltransferase 1/biosynthesis , Dose-Response Relationship, Drug , Epigenesis, Genetic/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Glucose/administration & dosage , Glucose/metabolism , Humans , Long Interspersed Nucleotide Elements , MutL Protein Homolog 1/biosynthesis , Oxidative Stress/genetics , Promoter Regions, Genetic , Reactive Oxygen Species/metabolism
5.
Curr Microbiol ; 75(4): 386-393, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29164291

ABSTRACT

Forty-four samples of traditional Doogh and yoghurt were collected from 13 regions of 4 provinces in west of Iran (13 area) and analyzed using molecular methods including PCR, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA, and sequencing. Moreover, collected samples as well as samples from industrially Doogh were analyzed with quantitative real-time PCR (RT-PCR). Analyzed 16S rRNA gene sequences of Doogh samples could be allocated to the presence of Lactobacillus spp. The typical yoghurt starter culture bacteria included four different Lactobacillus species with possible probiotic properties, L. acidophilus, L. helveticus, L. kefiranofaciens, and L. amylovorus. DGGE of traditional Doogh and yoghurt and RT-PCR of traditional Doogh and yoghurt and also industrial Doogh samples demonstrated that traditional Doogh and yoghurt show a higher abundance of total bacteria and lactobacilli and a higher bacterial diversity, respectively. Considering diversity and higher probiotic bacteria content in traditional Doogh, consumers' healthiness in tribes and villages could be promoted with these indigenous products.


Subject(s)
Cultured Milk Products/microbiology , Lactobacillus/isolation & purification , Yogurt/microbiology , Biodiversity , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , Food Microbiology , Iran , Lactobacillus/classification , Lactobacillus/genetics , Lactobacillus/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics
6.
Gene ; 537(1): 85-92, 2014 Mar 01.
Article in English | MEDLINE | ID: mdl-24325907

ABSTRACT

The human gut microbiota and microbial influences on lipid and glucose metabolism, satiety, and chronic low-grade inflammation are known to be involved in metabolic syndrome. Fermentation end products, especially short chain fatty acids, are believed to engage the epigenetic regulation of inflammatory reactions via FFARs (free fatty acid receptor) and other short chain fatty acid receptors. We studied a potential interaction of the microbiota with epigenetic regulation in obese and type 2 diabetes patients compared to a lean control group over a four month intervention period. Intervention comprised a GLP-1 agonist (glucagon-like peptide 1) for type 2 diabetics and nutritional counseling for both intervention groups. Microbiota was analyzed for abundance, butyryl-CoA:acetate CoA-transferase gene and for diversity by polymerase chain reaction and 454 high-throughput sequencing. Epigenetic methylation of the promoter region of FFAR3 and LINE1 (long interspersed nuclear element 1) was analyzed using bisulfite conversion and pyrosequencing. The diversity of the microbiota as well as the abundance of Faecalibacterium prausnitzii were significantly lower in obese and type 2 diabetic patients compared to lean individuals. Results from Clostridium cluster IV and Clostridium cluster XIVa showed a decreasing trend in type 2 diabetics in comparison to the butyryl-CoA:acetate CoA-transferase gene and according to melt curve analysis. During intervention no significant changes were observed in either intervention group. The analysis of five CpGs in the promoter region of FFAR3 showed a significant lower methylation in obese and type 2 diabetics with an increase in obese patients over the intervention period. These results disclosed a significant correlation between a higher body mass index and lower methylation of FFAR3. LINE-1, a marker of global methylation, indicated no significant differences between the three groups or the time points, although methylation of type 2 diabetics tended to increase over time. Our results provide evidence that a different composition of gut microbiota in obesity and type 2 diabetes affect the epigenetic regulation of genes. Interactions between the microbiota and epigenetic regulation may involve not only short chain fatty acids binding to FFARs. Therefore dietary interventions influencing microbial composition may be considered as an option in the engagement against metabolic syndrome.


Subject(s)
Diabetes Mellitus, Type 2/genetics , Epigenesis, Genetic , Fatty Acids, Volatile/metabolism , Gastrointestinal Tract/microbiology , Obesity/genetics , Receptors, G-Protein-Coupled/genetics , Adult , Aged , Biodiversity , Body Mass Index , Case-Control Studies , Coenzyme A-Transferases/genetics , DNA Methylation , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/microbiology , Feces/microbiology , Feeding Behavior , Female , Glucagon-Like Peptide 1/agonists , Glucagon-Like Peptide 1/analogs & derivatives , Glucagon-Like Peptide 1/therapeutic use , Gram-Positive Bacteria/physiology , Gram-Positive Endospore-Forming Bacteria/genetics , Humans , Liraglutide , Long Interspersed Nucleotide Elements , Male , Microbiota/physiology , Middle Aged , Obesity/microbiology , Promoter Regions, Genetic
7.
PLoS One ; 6(12): e28654, 2011.
Article in English | MEDLINE | ID: mdl-22194876

ABSTRACT

BACKGROUND: We investigated whether chemotherapy with the presence or absence of antibiotics against different kinds of cancer changed the gastrointestinal microbiota. METHODOLOGY/PRINCIPAL FINDINGS: Feces of 17 ambulant patients receiving chemotherapy with or without concomitant antibiotics were analyzed before and after the chemotherapy cycle at four time points in comparison to 17 gender-, age- and lifestyle-matched healthy controls. We targeted 16S rRNA genes of all bacteria, Bacteroides, bifidobacteria, Clostridium cluster IV and XIVa as well as C. difficile with TaqMan qPCR, denaturing gradient gel electrophoresis (DGGE) fingerprinting and high-throughput sequencing. After a significant drop in the abundance of microbiota (p = 0.037) following a single treatment the microbiota recovered within a few days. The chemotherapeutical treatment marginally affected the Bacteroides while the Clostridium cluster IV and XIVa were significantly more sensitive to chemotherapy and antibiotic treatment. DGGE fingerprinting showed decreased diversity of Clostridium cluster IV and XIVa in response to chemotherapy with cluster IV diversity being particularly affected by antibiotics. The occurrence of C. difficile in three out of seventeen subjects was accompanied by a decrease in the genera Bifidobacterium, Lactobacillus, Veillonella and Faecalibacterium prausnitzii. Enterococcus faecium increased following chemotherapy. CONCLUSIONS/SIGNIFICANCE: Despite high individual variations, these results suggest that the observed changes in the human gut microbiota may favor colonization with C. difficile and Enterococcus faecium. Perturbed microbiota may be a target for specific mitigation with safe pre- and probiotics.


Subject(s)
DNA Fingerprinting/methods , Denaturing Gradient Gel Electrophoresis/methods , Drug-Related Side Effects and Adverse Reactions , Feces/microbiology , Metagenome/genetics , Polymerase Chain Reaction/methods , Sequence Analysis, DNA/methods , Aged , Anti-Bacterial Agents/pharmacology , Bacteroides/drug effects , Bacteroides/genetics , Bifidobacterium/drug effects , Bifidobacterium/genetics , Case-Control Studies , Clostridium/drug effects , Clostridium/genetics , Clostridium/growth & development , Colony Count, Microbial , Genetic Variation , Health , High-Throughput Nucleotide Sequencing , Humans , Middle Aged , Neoplasms/genetics , Neoplasms/microbiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Taq Polymerase/metabolism
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