Screening efficient strains by cell platform is cost-effective, but to date, no screening experiments have been performed for targeted lactic acid bacteria with hypoxic/reoxygenation (H/R)-treated cardiomyocytes, and their effects on the phosphoinositide 3-kinase (PI3K)/protein kinase b (Akt)/endothelial nitric oxide synthase (eNOS) pathway in myocardial infarction (MI) are unclear. Here we activated 102 strains of lactic acid bacteria and inoculated them into MRS medium for fermentation. The fermentation supernatants of the lactic acid bacteria were incubated with an H/R model of H9C2 cells. We found that Bifidobacterium longum ZL0210 had the greatest potential for inhibiting the apoptosis of H/R-induced H9C2 cells. Furthermore, it significantly increased the expression of heme oxygenase-1 (HO-1) and quinone oxidoreductase 1 (NQO1) in H9C2 cardiomyocytes, as well as the Bcl-2/Bax protein ratio, protecting damaged myocardial cells via an anti-apoptotic pathway. Intragastric administration of B. longum ZL0210 to mice for one week before and after establishment of an MI model drastically attenuated the myocardial cell hypertrophy and fibrosis of the MI mice. Meanwhile, B. longum ZL0210 significantly reduced the secretion of myocardial enzymes, increased the activity of antioxidant enzymes, and inhibited lipid-oxidative malondialdehyde (MDA) levels. Moreover, it upregulated the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) protein and the phosphorylation levels of PI3K, Akt, and eNOS, resulting in increased NO contents. In summary, we screened 102 strains of lactic acid bacteria with a cell platform and determined that B. longum ZL0210 was a favorable candidate for protecting the myocardium, and we are the first to reveal the protective effects of B. longum ZL0210 for MI via activation of the PI3K/Akt/eNOS pathway through TRAIL.
Dysbiosis of gut microbiota has adverse effects on host health. This study aimed to determine the effects of changes of faecal microbiota in obese and diabetic rats on the imputed production of enzymes involved in the metabolism of glutamate, gamma-aminobutyric acid (GABA), and succinate. The levels of glutamate decarboxylase, GABA transaminase, succinate-semialdehyde dehydrogenase, and methylisocitrate lyase were reduced or absent in diabetic rats compared with controls and obese rats. Glutamate decarboxylase (GAD) was significantly reduced in obese rats compared with control rats, while the other enzymes were unaltered; different bacterial taxa are suggested to be involved. Levels of bacterial enzymes were inversely correlated with the blood glucose level. These findings suggest that the absence of GABA and reduced succinate metabolism from gut microbiota contribute to the diabetic state in rats.
The biological activities of acetic acid bacteria (AAB) as Gram-negative bacteria have attracted our interests, especially in their inhibitory effects on allergic responses. To clarify the underlying mechanism that improves allergic symptoms by ingestion of the AAB Gluconacetobacter hansenii,we examined whether different extracts of heat-killed G. hansenii GK-1 could reduce the IL-4 production of immune cells from food-allergic model of OVA23-3, transgenic mice with ovalbumin (OVA)-specific T-cell-receptor genes. A hot-water extract fraction (FII) of G. hansenii GK-1 significantly decreased the in vitro IL-4 production of spleen cells of OVA23-3 mice compared with those stimulated with OVA alone. The IL-4 inhibitory effect was also observed for FIV (purified LPS fraction), but the activity was lower than for FII or LPS from Escherichia coli. Unlike LPS from Escherichia coli, FIV significantly inhibited the LPS-induced IL-6 production of the spleen cells. The addition of FII or FIV to a Foxp3+T cell-inducing culture showed that FII significantly promoted the rate of Foxp3+CD4+T cells of OVA-stimulated mesenteric lymph node cells from recombination-activating-gene (RAG)-2-deficient food-allergic inflammatory OVA23-3 (R23-3) mice with suppression of IL-4 production, while FIV induced Foxp3+T cells from RAG-2-deficient DO11.10 non-inflammatory mice. Structure analysis showed a lack of O-antigen in FIV, which seemed to lead to the weak biological activities of FIV observed. The present study suggests that extracts of G. hansenii GK-1 to inhibit IL-4 production of immune cells and/or promote regulatory T cell differentiation synergistically play important roles in improving allergic symptoms safely as well as normal condition.
Lactococcus lactis subsp. Cremoris C60 is a probiotic strain that induces diverse functional modifications in immune cells. In this report, as a novel effect of C60 on myeloid lineage cells, we show that C60 enhances the immunological function of macrophages that consequently promotes CD4+ T cell activity in an antigen-dependent manner. Heat-killed (HK) C60 induced the production of pro-inflammatory cytokines in thioglycolate-elicited peritoneal macrophages (TPMs) much stronger than Toll-like receptor (TLR) ligand stimulation. The HK-C60 treatment also augmented the expression of antigen-presenting and co-stimulatory molecules, such as major histocompatibility complex (MHC) class II, CD80, and CD86, as well as antigen uptake in TPMs. These HK-C60-mediated functional upregulations in TPMs resulted in the promotion of CD4+ T cell activation in an antigen-dependent manner. Interestingly, the TPMs that originated from the mice fed the HK-C60 diet showed pre-activated characteristics, which was confirmed by the upregulation of cytokine production and antigen presentation-related molecule expression under lipopolysaccharide (LPS) stimulation. Furthermore, the antigen-dependent CD4+ T cell activation was also enhanced by the TPMs. This implied that antigen presentation activity was enhanced in the TPMs that originated from the HK-C60 diet mice. Thus, C60 effectively upregulates the immunological function of macrophages that directly connects to CD4+ T cell-based adaptive immunity.
Intestinal parasitic infections can change gut microbiota and short-chain fatty acids (SCFAs). We aimed to study the interaction among Strongyloides stercoralis, human gut microbiota, and serum SCFAs in a community. Fifty-two subjects in Donchang sub-district, Khon Kaen Province, northeastern Thailand, were included based on specific inclusion and exclusion criteria. Characteristics of the participants were matched between those positive for S. stercoralis infection alone (no other intestinal parasites; Ss+, n=26) and uninfected controls (infection status confirmed by PCR; Ss−, n=26). Serum short-chain fatty acids were evaluated by gas chromatography-mass spectrometry. DNA was extracted from individual faecal samples and then pooled into two groups (Ss+ and Ss−) for amplification and sequencing of the V3-V4 region of the 16S gene with next-generation technology. We explored the impact of infection with S. stercoralis on the faecal microbiota: individuals infected with this parasite exhibited increased alpha diversity of bacteria. At the genus level, gut microbiota in Ss+ patients showed high abundances of Escherichia-Shigella and Bacteroides but low abundances of the genera Bifidobacterium, Lactobacillus, and Blautia. PCR of individual samples to identify certain species of interest gave results consistent with those from next-generation sequencing of pooled samples and showed that significantly more Ss+ samples contained Bacteroides fragilis. Intriguingly, a major SCFA, acetic acid, was significantly decreased in S. stercoralis infection.In conclusion, S. stercoralis infection caused an imbalance of gut microbiota and decreased acetic acid in serum. This information adds to the knowledge concerning the effect of intestinal nematode-related chronic diseases.
On March 11, 2020, the World Health Organization declared a pandemic of coronavirus infectious disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and imposed the biggest public health challenge for our civilization, with unforeseen impacts in the subsequent years. Similar to other respiratory infections, COVID-19 is associated with significant changes in the composition of the upper respiratory tract microbiome. Studies have pointed to a significant reduction of diversity and richness of the respiratory microbiota in COVID-19 patients. Furthermore, it has been suggested that Prevotella, Staphylococcus, and Streptococcus are associated with severe COVID-19 cases, while Dolosigranulum and Corynebacterium are significantly more abundant in asymptomatic subjects or with mild disease. These results have stimulated the search for new microorganisms from the respiratory microbiota with probiotic properties that could alleviate symptoms and even help in the fight against COVID-19. To date, the potential positive effects of probiotics in the context of SARS-CoV-2 infection and COVID-19 pandemics have been extrapolated from studies carried out with other viral pathogens, such as influenza virus and respiratory syncytial virus. However, scientific evidence has started to emerge demonstrating the capacity of immunomodulatory bacteria to beneficially influence the resistance against SARS-CoV-2 infection. Here we review the scientific knowledge regarding the role of the respiratory microbiota in viral infections in general and in the infection caused by SARS-CoV-2 in particular. In addition, the scientific work that supports the use of immunomodulatory probiotic microorganisms as beneficial tools to reduce the severity of respiratory viral infections is also reviewed. In particular, our recent studies that evaluated the role of immunomodulatory Dolosigranulum pigrum strains in the context of SARS-CoV-2 infection are highlighted.
Eight bacterial strains were used in this study to examine the survival of intestinal bacteria in immune cell cultures under aerobic and anaerobic culture conditions. With the addition of penicillin G and streptomycin, viable Clostridium clostridioforme and Fusobacterium varium cells did not decrease after 6 or 24 hours, even under aerobic conditions. Without antibiotics, eight bacterial strains did not decrease until 4 or 6 hours later, under both aerobic and anaerobic conditions. Escherichia coli numbers increased by more than 10 times under both conditions. In order to examine the effects of live gut bacteria on various immune cells, the viability of bacteria should be checked in cell culture media and under different conditions.
Recent research has confirmed that moderate-intensity exercise affects the gut microbiome composition and improves cardiac function in an animal model after myocardial infarction (MI). However, few studies have investigated the effects of exercise on glucose and lipid metabolism in patients with coronary heart disease (CHD) receiving a statin treatment and successful percutaneous coronary intervention (PCI). Meanwhile, since statin therapy may lead to the risk of an increase in blood glucose level in CHD patients, we hypothesized that moderate-intensity exercise may be helpful for regulating glucose-lipid metabolism and stabilizing the blood glucose level in CHD patients. Therefore, to confirm our conjecture, we conducted a clinical retrospective study and animal experiment, respectively. The clinical study involved a total of 501 statin-treated patients with CHD after PCI. According to the study protocol, patients were divided into the following three groups: a non-exercise group, exercise at the recommended standard group, and exercise not at the recommended standard group. We found that qualified moderate-intensity exercise decreased blood glucose and lipid levels at follow-up at a mean of 2.2 years, and the incidence of new-onset diabetes showed a downward trend compared with the non-exercise and exercise not at the recommended standard groups. Furthermore, we used a high-fat rat model to explore an additional mechanism of the beneficial effects of exercise-based management on glucose-lipid metabolism apart from the known mechanism. We used 16S rRNA high-throughput sequencing technology to analyze the changes induced by exercise in the composition of intestinal flora in experimental rats. We found that rats that exercised with or without statin administration had lower plasma glucose and lipid levels and that these parameters were higher in the control and statin-treated rats that did not exercise. These results were consistent with the human study. The results from high-throughput sequencing of the intestinal flora of rats showed, to the best of our knowledge, that exercise leads to an increased relative abundance of Akkermansia muciniphila, which contributes to improved glucose and lipid metabolism. Based on our current results, we suggest that moderate-intensity exercise can improve glucose and lipid metabolism and prevent statin treatment-related side effects, such as hyperglycemia, in patients after PCI. Exercise could facilitate the applicability of statins for lower lipid levels. Exercise training also provides additional benefits, such as alteration of the gut microbiota, which contributes to improved glucose and lipid metabolism.
Chronic inflammation caused by gut dysbiosis is associated with the pathophysiology of metabolic disease. Synbiotics are useful for ameliorating gut dysbiosis; however, it remains unclear what types of bacteria act as key markers for synbiotic-driven improvement of chronic inflammation. Here, we performed a post hoc analysis of a 24-week randomized controlled study using synbiotics to investigate the association between gut microbiota and inflammatory markers. We characterized the responders who showed lower interleukin-6 (IL-6) levels in response to synbiotic supplementation among 86 obese patients with type 2 diabetes mellitus. In our baseline analysis, the relative abundances of Bifidobacterium adolescentis and Alistipes onderdonkii correlated positively with IL-6, lipopolysaccharide binding protein (LBP), and high-sensitivity C-reactive protein (Hs-CRP) levels. The relative abundance of Eubacterium rectale correlated positively with LBP and Hs-CRP levels, and that of Bacteroides thetaiotaomicron correlated positively with LBP levels. Based on our responder analysis, patients with higher body mass indices (over 30 kg/m2 on average), low abundances of Bacteroides caccae and Parabacteroides merdae at baseline and 24 weeks, and minimal changes in the relative abundance of E. rectale and Shannon index from baseline showed decreased IL-6 levels compared with baseline. However, glycemic control in responders was unchanged. In conclusion, we identified four bacterial species (B. adolescentis, A. onderdonkii, E. rectale, and B. thetaiotaomicron)related to chronic inflammation and predictive markers (B. caccae,P. merdae, and severity of obesity) in responders to synbiotic supplementation among obese patients with type 2 diabetes.
The increase of lifestyle-related diseases in Asia has recently become remarkably serious. This has been associated with a change in dietary habits that may alter the complex gut microbiota and its metabolic function in Asian people. Notably, the penetration of modern Western diets into Asia, which has been accompanied by an increase in fat content and decrease in plant-derived dietary fiber, is restructuring the Asian gut microbiome. In this review, we introduce the current status of obesity and diabetes in Asia and discuss the links of changes in dietary style with gut microbiota alterations which may predispose Asian people to metabolic diseases.
Probiotics are considered effective microbial dietary supplements that provide beneficial effects to consumers, usually by restoring or improving gut microflora. Goat milk is one of the rich sources of probiotics as well as nutrients. Therefore, the primary aim of this research was to isolate and evaluate the potentials of novel indigenous probiotic strains present in goat milk. Six different raw goat milk samples were collected from different areas of Multan, Pakistan. For bacterial characterization, samples were cultured and isolated on MRS agar plates for different morphological and biochemical tests. The probiotic potentials of the six isolates, all of which were gram positive (G1, G2, G3, G4, G5, and G6) and five of which were catalase negative (all except G1), were assessed via a milk coagulation assay and antimicrobial activity, pH tolerance, phenol tolerance, and sodium chloride (NaCl) tolerance tests, which revealed that all the isolates coagulated in milk and showed protease and lipase activity, except G3. All six isolates showed tolerance against 0.2% phenol and 2–4% NaCl and were able to survive in both alkaline and acidic conditions. Only five isolates showed antimicrobial activity against indicator strain Aspergillusniger strain STA9, validating their probiotic nature. The most potent bile-tolerant and bacteriocin-producing isolate, G1, also showed γ-hemolytic activity and resistance to penicillin but showed susceptibility to other antibiotics. The lactic acid-producing (0.60% titratable acidity) G1 isolate was identified as a novel strain of Mammaliicoccussciuri based on 16S rDNA sequencing. The above findings suggest that the potent Mammaliicoccussciuri GMN01 strain can serve as a potential probiotic strain. A potent probiotic strain isolated from raw goat milk could be utilized as a dietary supplement, and goat milk could become an alternative to other sources of milk, particularly cow milk. However, safety aspects of this strain require further investigation because the present safety tests are insufficient to conclude that the GMN01 isolate is safe.