Intensive Training and Sex Influence Intestinal Microbiota Composition: A Preclinical Approach

Lifestyle, including regular physical activity and dietary habits, influences microbiota composition. Although some studies have demonstrated changes in microbiota composition due to moderate or high intensity training in athletes, the relationship between physical activity and changes in the intestinal bacteria is still a matter of discussion. In addition, as most studies are performed on males, the role of sexual dimorphism deserves to be explored. Therefore, the aim of this preliminary study was to ascertain the influence of an intensive training and the rat's sex on caecal microbiota composition. For this purpose, female and male 4-week-old Wistar rats were submitted to a 4-week running training program. At the end, caecal samples were collected immediately after performing an exhaustion test to characterize the microbiota composition by 16S rRNA sequencing technique. Parallel groups of female and male sedentary (SED) rats (age matched) were included. The results showed that young female rats had a higher ability to run than males but no sex- or training-associated changes were observed on microbiota diversity and richness indexes among groups. However, the Actinobacteria, Bifidobacteriaceae and Bifidobacterium spp. proportions were significantly higher in male than in female rats when comparing SED groups (p <0.05), whose proportions in males were decreased by the training program (p < 0.05 vs. SED). On the contrary, female SED rats showed a higher proportion of Odoribacteraceae (belonging to Bacteroidetes), Clostridiaceae and Eubacteriaceae (both Firmicutes) than in the respective male group (p < 0.05), although Eubacteriaceae proportion decreased by running (p < 0.05 vs. SED). However, training increased the proportion of the Paraeggerthella genus (Actinobacteria) in female rats with respect to its sedentary counterpart (p < 0.05). Overall, caecal microbiota composition is modified by intensive training in young rats but there are also sex-based differences that should be considered for interventional studies

Alterations in the innate immune system due to exhausting exercise in intensively trained rats

It is known that intensive physical activity alters the immune system's functionality. However, the influence of the intensity and duration of exercise needs to be studied in more depth. We aimed to establish the changes in the innate immune response induced by two programmes of intensive training in rats compared to sedentary rats. A short training programme included 2 weeks of intensive training, ending with an exhaustion test (short training with exhaustion, S-TE). A second training programme comprised 5-week training including two exhaustion tests and three trainings per week. In this case, immune status was assessed before (T), immediately after (TE) and 24 h after (TE24) an additional final exhaustion test. Biomarkers such as phagocytic activity, macrophage cytokine and reactive oxygen species (ROS) production, and natural killer (NK) cell activity were quantified. S-TE was not enough to induce changes in the assessed innate immunity biomarkers. However, the second training was accompanied by a decrease in the phagocytic activity, changes in the pattern of cytokine secretion and ROS production by macrophages and reduced NK cell proportion but increased NK cytotoxic activity. In conclusion, a 5-week intense training programme, but not a shorter training, induced alterations in the innate immune system functionality

Alteractions in the mucosal immune system by a chronic exhausting exercise in Wistar rats

Exhausting exercise can disturb immune and gastrointestinal functions. Nevertheless, the impact of it on mucosal-associated lymphoid tissue has not been studied in depth. Here, we aim to establish the effects of an intensive training and exhausting exercise on the mucosal immunity of rats and to approach the mechanisms involved. Rats were submitted to a high-intensity training consisting of running in a treadmill 5 days per week for 5 weeks, involving 2 weekly exhaustion tests. At the end, samples were obtained before (T), immediately after (TE) and 24 h after (TE24) an additional final exhaustion test. The training programme reduced the salivary production of immunoglobulin A, impaired the tight junction proteins' gene expression and modified the mesenteric lymph node lymphocyte composition and function, increasing the ratio between Tαβ+ and B lymphocytes, reducing their proliferation capacity and enhancing their interferon-γ secretion. As a consequence of the final exhaustion test, the caecal IgA content increased, while it impaired the gut zonula occludens expression and enhanced the interleukin-2 and interferon-γ secretion. Our results indicate that intensive training for 5 weeks followed or not by an additional exhaustion disrupts the mucosalassociated lymphoid tissue and the intestinal epithelial barrier integrity in rats

Hesperidin Effects on Gut Microbiota and Gut-Associated Lymphoid Tissue in Healthy Rats

Hesperidin, found in citrus fruits, has shown a wide range of biological properties. Nonetheless, a more in-depth investigation is required on the effects on the immune system, and in particular, on the gut-associated lymphoid tissue, together with its relationship with the gut microbiota. Therefore, we aimed to establish the influence of oral hesperidin administration on the intestinal lymphoid tissue and on the gut microbiota composition in healthy animals. Lewis rats were orally administrated 100 or 200 mg/kg hesperidin three times per week for four weeks. Microbiota composition and IgA-coated bacteria were determined in caecal content. Mesenteric lymph node lymphocyte (MLNL) composition and functionality were assessed. IgA, cytokines, and gene expression in the small intestine were quantified. Hesperidin administration resulted in a higher number of bacteria and IgA-coated bacteria, with changes in microbiota composition such as higher Lactobacillus proportion. Hesperidin was also able to increase the small intestine IgA content. These changes in the small intestine were accompanied by a decrease in interferon-γ and monocyte chemotactic protein-1 concentration. In addition, hesperidin increased the relative proportion of TCRαβ+ lymphocytes in MLNL. These results show the immunomodulatory actions of hesperidin on the gut-associated lymphoid tissue and reinforce its role as a prebiotic. KEYWORDS: flavanone; flavonoids; immunoglobulin A; intestinal immunity; polyphenol; prebioti

Changes in intestinal immune system after training and exhausting physical exercise in rats

Podeu consultar el III Workshop anual INSA-UB complet a: http://hdl.handle.net/2445/118993Sessió 1. Pòster núm.

Changes in lymphocyte composition and functionality after intensive training and exhausting exercise in rats

Exhausting exercise can have a deleterious effect on the immune system. Nevertheless, the impact of exercise intensity on lymphocyte composition and functionality remains uncertain. The aim of this study was to establish the influence of intensive training on lymphoid tissues (blood, thymus, and spleen) in Wistar rats. Two intensive training programs were performed: a short program, running twice a day for 2 weeks and ending with a final exhaustion test (S-TE group), and a longer program, including two exhaustion tests plus three runs per week for 5 weeks. After this last training program, samples were obtained 24 h after a regular training session (T group), immediately after an additional exhaustion test (TE group) and 24 h later (TE24 group). The composition of lymphocytes in the blood, thymus, and spleen, the function of spleen cells and serum immunoglobulins were determined. In the blood, only the TE group modified lymphocyte proportions. Mature thymocytes' proportions decreased in tissues obtained just after exhaustion. There was a lower percentage of spleen NK and NKT cells after the longer training program. In these rats, the T group showed a reduced lymphoproliferative activity, but it was enhanced immediately after the final exhaustion. Cytokine secretion was modified after the longer training (T group), which decreased IFN-g and IL-10 secretion but increased that of IL-6. Higher serum IgG concentrations after the longer training program were detected. In conclusion, the intensive training for 5 weeks changed the lymphocyte distribution among primary and secondary lymphoid tissues and modified their function

Influence of hesperidin on systemic immunity of rats following an intensive training and exhausting exercise

Intensive training and exhausting exercise can disrupt innate and acquired immunity. The flavanone hesperidin has shown immunomodulatory properties in physiological and some pathological conditions, and positive e ects on exercise-induced oxidative stress. Nevertheless, it remains uncertain whether it also prevents exhausting exercise-induced immune alterations. The aim of this study was to establish the e ect of oral hesperidin supplementation on the systemic immune system in rats following an intensive training and exhausting exercise. For this purpose, female Wistar rats were randomized into an intensive training group or a sedentary group. Intensive training was induced by running in a treadmill 5 days per week (including two exhausting tests) for five weeks. Throughout the training period, 200 mg/kg of hesperidin or vehicle was administered by oral gavage three times per week. At the end, blood, thymus, spleen and macrophages were collected before, immediately after and 24 h after an additional final exhaustion test. Hesperidin supplementation enhanced natural killer cell cytotoxicity and the proportion of phagocytic monocytes, attenuated the secretion of cytokines by stimulated macrophages, prevented the leukocytosis induced by exhaustion and increased the proportion of T helper cells in the thymus, blood and spleen. These results suggest that hesperidin can prevent exhausting exercise-induced immune alteration

Protective Effect of Hesperidin on the Oxidative Stress Induced by an Exhausting Exercise in Intensively Trained Rats

Intensive exercise can lead to oxidative stress, which can be particularly deleterious for lymphoid tissues. Hesperidin has demonstrated its antioxidant activity, but few studies focus on its influence on intensive training. The aim of this study was to assess the impact of hesperidin on the oxidant/antioxidant status of lymphoid tissues after an intensive training program. Wistar rats were trained for five weeks (five days per week), including two exhaustion tests plus three trainings per week. During this period, animals were orally administrated with 200 mg/kg of hesperidin or vehicle (three days per week). The oxidative status was determined before, immediately after and 24 h after an additional exhaustion test. The production of reactive oxygen species (ROS) by peritoneal macrophages, superoxide dismutase (SOD) and catalase activities in spleen, thymus and liver, and hepatic glutathione peroxidase activity (GPx) were assessed. Hesperidin prevented an increase in ROS production induced by the additional exhaustion test. Likewise, hesperidin avoided a decrease in SOD and catalase activities in the thymus and spleen that was found after the additional exhaustion test. The antioxidant effects of hesperidin were associated with a higher performance in the assessed training model. These results suggest that hesperidin, acting as an antioxidant, can prevent oxidative stress induced by exercise and improve exercise performance

Biomarkers of legume intake in human intervention and observational studies: A systematic review

There is a growing interest in assessing dietary intake more accurately across different population groups, and biomarkers have emerged as a complementary tool to replace traditional dietary assessment methods. The purpose of this study was to conduct a systematic review of the literature available and evaluate the applicability and validity of biomarkers of legume intake reported across various observational and intervention studies. A systematic search in PubMed, Scopus, and ISI Web of Knowledge identified 44 studies which met the inclusion criteria for the review. Results from observational studies focused on soy or soy-based foods and demonstrated positive correlations between soy intake and urinary, plasma or serum isoflavonoid levels in different population groups. Similarly, intervention studies demonstrated increased genistein and daidzein levels in urine and plasma following soy intake. Both genistein and daidzein exhibited dose-response relationships. Other isoflavonoid levels such as O-desmethylangolensin (O-DMA) and equol were also reported to increase following soy consumption. Using a developed scoring system, genistein and daidzein can be considered as promising candidate markers for soy consumption. Furthermore, genistein and daidzein also served as good estimates of soy intake as evidenced from long-term exposure studies marking their status as validated biomarkers. On the contrary, only few studies indicated proposed biomarkers for pulses intake, with pipecolic acid and S-methylcysteine reported as markers reflecting dry bean consumption, unsaturated aliphatic, hydroxyl-dicarboxylic acid related to green beans intake and trigonelline reported as marker of peas consumption. However, data regarding criteria such as specificity, dose-response and time-response relationship, reliability, and feasibility to evaluate the validity of these markers is lacking. In conclusion, despite many studies suggesting proposed biomarkers for soy, there is a lack of information on markers of other different subtypes of legumes. Further discovery and validation studies are needed in order to identify reliable biomarkers of legume intake

Discovery of intake biomarkers of lentils, chickpeas and white beans by untargeted LC-MS metabolomics in serum and urine.

Scope: To identify reliable biomarkers of food intake (BFIs) of pulses. Methods and results: A randomized crossover postprandial intervention study is conducted on 11 volunteers who consumed lentils, chickpeas, and white beans. Urine and serum samples are collected at distinct postprandial time points up to 48 h, and analyzed by LC-HR-MS untargeted metabolomics. Hypaphorine, trigonelline, several small peptides, and polyphenol-derived metabolites prove to be the most discriminating urinary metabolites. Two arginine-related compounds, dopamine sulfate and epicatechin metabolites, with their microbial derivatives, are identified only after intake of lentils, whereas protocatechuic acid is identified only after consumption of chickpeas. Urinary hydroxyjasmonic and hydroxydihydrojasmonic acids, as well as serum pipecolic acid and methylcysteine, are found after white bean consumption. Most of the metabolites identified in the postprandial study are replicated as discriminants in 24 h urine samples, demonstrating that in this case the use of a single, noninvasive sample is suitable for revealing the consumption of pulses. Conclusions: The results of the present untargeted metabolomics work reveals a broad list of metabolites that are candidates for use as biomarkers of pulse intake. Further studies are needed to validate these BFIs and to find the best combinations of them to boost their specificity