Protective effects of SKLB023 on a mouse model of unilateral ureteral obstruction by the modulation of gut microbiota

Renal interstitial fibrosis is the common pathway underlying the progression of chronic kidney disease (CKD) to end-stage renal disease (ESRD) and the corresponding therapies are limited. Quantitative and qualitative alterations in gut microbiota are noted in patients with CKD and ESRD. In our previous study, SKLB023 exhibited antifibrotic effects by interfering TGF-β1/Smad3 signaling in obstructive nephropathy. However, it remained unclear that oral administration of SKLB023 drives the alteration of gut microbiota to attenuate renal fibrosis. In the study, the marked inflammation and interstitial fibrosis were found in the kidney tissues of unilateral ureteral obstruction (UUO) mice. While treatment with SKLB023 significantly alleviated renal interstitial fibrosis and reduced serum proinflammatory cytokines TNF-α, IL-6 levels. Importantly, SKLB023 derived the modulation of gut microbiota with the increasing similarity between the composition of gut microbiota in the control and UUO. The number of Turicibacter and Candidatus_Arthromitus was significantly decreased following UUO surgery and recovered by SKLB023, which positively correlated with pro-inflammatory cytokine expression. These results indicated the potential relationship between the antifibrotic benefits of SKLB023 and gut microbiota alteration, which provided new insights into drug therapy via gut microbiota modulation in obstructive nephropathy.

Renal fibrosis is the common pathway underlying the progression of CKD to ESRD and quantitative and qualitative alterations in gut microbiota are noted in patients with CKD. Our results indicated SKLB023 drives the alteration of gut microbiota to attenuate renal fibrosis.     

Plasma Lipopolysaccharide Is Closely Associated With Glycemic Control and Abdominal Obesity: Evidence from bariatric surgery

OBJECTIVE

It is of vital importance to elucidate the triggering factors of obesity and type 2 diabetes to improve patient care. Bariatric surgery has been shown to prevent and even cure diabetes, but the mechanism is unknown. Elevated levels of lipopolysaccharide (LPS) predict incident diabetes, but the sources of LPS are not clarified. The objective of the current study was to evaluate the potential impact of plasma LPS on abdominal obesity and glycemic control in subjects undergoing bariatric surgery.

RESEARCH DESIGN AND METHODS

This was a prospective observational study involving a consecutive sample of 49 obese subjects undergoing bariatric surgery and 17 controls. Main assessments were plasma LPS, HbA_1c, adipose tissue volumes (computed tomography), and quantified bacterial DNA in adipose tissue compartments.

RESULTS

Plasma levels of LPS were elevated in obese individuals compared with controls (P < 0.001) and were reduced after bariatric surgery (P = 0.010). LPS levels were closely correlated with HbA_1c (r = 0.56; P = 0.001) and intra-abdominal fat volumes (r = 0.61; P < 0.001), but only moderately correlated with subcutaneous fat volumes (r = 0.33; P = 0.038). Moreover, there was a decreasing gradient (twofold) in bacterial DNA levels going from mesenteric via omental to subcutaneous adipose tissue compartments (P = 0.041). Finally, reduced LPS levels after bariatric surgery were directly correlated with a reduction in HbA_1c (r = 0.85; P < 0.001).

CONCLUSIONS

Our findings support a hypothesis of translocated gut bacteria as a potential trigger of obesity and diabetes, and suggest that the antidiabetic effects of bariatric surgery might be mechanistically linked to, and even the result of, a reduction in plasma levels of LPS.Obesity and type 2 diabetes are rapidly emerging as major public health problems worldwide. This growing pandemic is often associated with other prevalent diseases, such as insulin resistance, metabolic syndrome, and cardiovascular disease, which are disease states that are potentially linked to chronic low-grade inflammation (1). Obesity per se facilitates a proinflammatory state, characterized by increased levels of proinflammatory cytokines, and it has been proposed that adipose tissue, particularly intra-abdominal adipose tissue, might be a major source of inflammation (2,3). However, the triggering factors of adipose tissue inflammation, obesity, and type 2 diabetes remain to be determined.Interestingly, mesenteric fat, which is localized in close proximity to the gut wall, in mice has been shown to express higher levels of proinflammatory chemokines than other types of adipose tissue (4). The gut flora or microbiota contains 10-fold the number of cells and 150-times the number of genes compared with the human body and is currently being characterized in the Human Microbiome Project and in the MetaHIT program (5). An altered gut microbiota has been linked to several chronic disease states, including obesity (6) and type 2 diabetes (7).Translocation of gut microbiota, particularly endotoxins or lipopolysaccharides (LPSs) on the surface of gram-negative bacteria to the systemic circulation, has been proposed to be an early trigger of inflammation and obesity (8). Endotoxins normally circulate at low levels in healthy individuals (9), and 80–97% of circulating endotoxin is bound to lipoproteins (10). LPS promotes inflammation mainly by signaling through Toll-like receptor (TLR) 4 on macrophages, monocytes, and other cells of the innate immune system, and CD14 plays a central role by transferring LPS to the TLR4 complex (11).An increase in plasma LPS occurs in healthy individuals after a high-fat meal (12), whereas a chronic state of low-grade endotoxemia as measured by plasma LPS (8) or LPS-binding protein (13) is evident in patients with obesity and insulin resistance. Furthermore, low-grade endotoxemia predicts incident diabetes, but the sources of LPS are unknown because LPS can translocate both from the oral cavity and the gut (14). Interestingly, in a mouse model, commensal gut bacteria translocate to mesenteric adipose tissue, initiating low-grade inflammation before the onset of insulin resistance and type 2 diabetes (15). To our knowledge, these mechanisms have not been studied in humans.Bariatric surgery has been shown to prevent and potentially cure diabetes (16–18), but the mechanism is unknown. We hypothesized that adipose tissue in proximity to the gut would have a relatively higher quantity of bacterial DNA than other types of adipose tissue, that circulating plasma LPS would be associated with the amount of intra-abdominal adipose tissue and HbA_1c, and that a reduction of LPS after bariatric surgery would correlate with improved glycemic control.     

Structural characterization of phosphoethanolamine-modified lipid A from probiotic Escherichia coli strain Nissle 1917

Gut microbiota, a complex microbial community inhabiting human or animal intestines recently regarded as an endocrine organ, has a significant impact on human health. Probiotics can modulate gut microbiota and the gut environment by releasing a range of bioactive compounds. Escherichia coli (E. coli) strain Nissle 1917 (EcN), a Gram-negative bacterial strain, has been used to treat gastrointestinal (GI) disorders (i.e., inflammatory bowel disease, diarrhea, ulcerative colitis, and so on). However, endotoxicity of lipopolysaccharide (LPS), a major component of the cell wall of Gram-negative bacteria in the gut, is known to have a strong influence on gut inflammation and maintenance of gut homeostasis. Therefore, characterizing the chemical structure of lipid A which determines the toxicity of LPS is needed to understand nonpathogenic colonization and commensalism properties of EcN in the gut more precisely. In the present study, MALDI multiple-stage mass spectrometry analysis of lipid A extracted from EcN demonstrates that hexaacylated lipid A (m/z 1919.19) contains a glucosamine disaccharide backbone, a myristate, a laurate, four 3-hydroxylmyristates, two phosphates, and phosphoethanolamine (PEA). PEA modification of lipid A is known to contribute to cationic antimicrobial peptide (CAMP) resistance of Gram-negative bacteria. To confirm the role of PEA in CAMP resistance of EcN, minimum inhibitory concentrations (MICs) of polymyxin B and colistin were determined using a wild-type strain and a mutant strain with deletion of eptA gene encoding PEA transferase. Our results confirmed that MICs of polymyxin B and colistin for the wild-type were twice as high as those for the mutant. These results indicate that EcN can more efficiently colonize the intestine through PEA-mediated tolerance despite the presence of CAMPs in human gut such as human defensins. Thus, EcN can be used to help treat and prevent many GI disorders.

The probiotic Escherichia coli strain Nissle 1917 is able to colonize the gut more efficiently through lipid A mediated cationic antimicrobial peptide tolerance.     

Lipid- and gut microbiota-modulating effects of graphene oxide nanoparticles in high-fat diet-induced hyperlipidemic mice

Graphene oxide (GO) suspensions can act as a good dispersant and drug delivery system for effective dispersion and drug sustained release. In this study, we investigated the impact of GO on blood/liver lipids and gut microbiota structure in high-fat diet (HFD)-induced hyperlipidemic mice. Oral administration of GO for 28 days remarkably decreased the lipid levels in blood and liver. GO did not decrease the total number of gut bacteria but increased the relative abundance of short-chain fatty acid (SCFA)-producing bacteria such as Clostridium clusters IV and Allobaculum spp. GO also enhanced the copying of bacterial butyryl coenzyme A transferase (BcoA), a key butyrate-producing gene. Although further pharmacological studies are still needed, these results provided an interesting hint that GO may exert beneficial effects on the host''s metabolism via selective modulation of SCFA-producing gut microbes.

The impact of GO on blood/liver lipids and gut microbiota structure in high-fat diet (HFD)-induced hyperlipidemic mice was investigated. Oral administration of GO for 28 days remarkably decreased lipid levels in blood and liver and GO did not decrease the total number of gut bacteria but increased the relative abundance of short-chain fatty acid (SCFA)-producing bacteria.     

Regulation of Serum Amyloid A3 (SAA3) in Mouse Colonic Epithelium and Adipose Tissue by the Intestinal Microbiota

The gut microbiota has been proposed as an environmental factor that affects the development of metabolic and inflammatory diseases in mammals. Recent reports indicate that gut bacteria-derived lipopolysaccharide (LPS) can initiate obesity and insulin resistance in mice; however, the molecular interactions responsible for microbial regulation of host metabolism and mediators of inflammation have not been studied in detail. Hepatic serum amyloid A (SAA) proteins are markers and proposed mediators of inflammation that exhibit increased levels in serum of insulin-resistant mice. Adipose tissue-derived SAA3 displays monocyte chemotactic activity and may play a role in metabolic inflammation associated with obesity and insulin resistance. To investigate a potential mechanistic link between the intestinal microbiota and induction of proinflammatory host factors, we performed molecular analyses of germ-free, conventionally raised and genetically modified Myd88−/− mouse models. SAA3 expression was determined to be significantly augmented in adipose (9.9±1.9-fold; P<0.001) and colonic tissue (7.0±2.3-fold; P<0.05) by the presence of intestinal microbes. In the colon, we provided evidence that SAA3 is partially regulated through the Toll-like receptor (TLR)/MyD88/NF-kappaB signaling axis. We identified epithelial cells and macrophages as cellular sources of SAA3 in the colon and found that colonic epithelial expression of SAA3 may be part of an NF-kappaB-dependent response to LPS from gut bacteria. In vitro experiments showed that LPS treatments of both epithelial cells and macrophages induced SAA3 expression (27.1±2.5-fold vs. 1.6±0.1-fold, respectively). Our data suggest that LPS, and potentially other products of the indigenous gut microbiota, might elevate cytokine expression in tissues and thus exacerbate chronic low-grade inflammation observed in obesity.     

Obesity,Diabetes, and Gut Microbiota: The hygiene hypothesis expanded?

The connection between gut microbiota and energy homeostasis and inflammation and its role in the pathogenesis of obesity-related disorders are increasingly recognized. Animals models of obesity connect an altered microbiota composition to the development of obesity, insulin resistance, and diabetes in the host through several mechanisms: increased energy harvest from the diet, altered fatty acid metabolism and composition in adipose tissue and liver, modulation of gut peptide YY and glucagon-like peptide (GLP)-1 secretion, activation of the lipopolysaccharide toll-like receptor-4 axis, and modulation of intestinal barrier integrity by GLP-2. Instrumental for gut microbiota manipulation is the understanding of mechanisms regulating gut microbiota composition. Several factors shape the gut microflora during infancy: mode of delivery, type of infant feeding, hospitalization, and prematurity. Furthermore, the key importance of antibiotic use and dietary nutrient composition are increasingly recognized. The role of the Western diet in promoting an obesogenic gut microbiota is being confirmation in subjects. Following encouraging results in animals, several short-term randomized controlled trials showed the benefit of prebiotics and probiotics on insulin sensitivity, inflammatory markers, postprandial incretins, and glucose tolerance. Future research is needed to unravel the hormonal, immunomodulatory, and metabolic mechanisms underlying microbe-microbe and microbiota-host interactions and the specific genes that determine the health benefit derived from probiotics. While awaiting further randomized trials assessing long-term safety and benefits on clinical end points, a healthy lifestyle—including breast lactation, appropriate antibiotic use, and the avoidance of excessive dietary fat intake—may ensure a friendly gut microbiota and positively affect prevention and treatment of metabolic disorders.Along with the increasing worldwide incidence of obesity-associated disorders, research has recently unraveled important pathways reciprocally connecting metabolism with the immune system. The development of obesity is a complex process involving genetic susceptibility and environmental factors, which both remain only partially understood. In such instances, gut microbiota is being increasingly recognized as an important factor connecting genes, environment, and immune system. The human gut hosts an enormous number and variety of microorganisms, including at least 10¹⁴ bacteria belonging to ∼1,000 species (1). The genome size of this microbial organ, collectively named microbiome, exceeds the size of the human nuclear genome by two orders of magnitude and provides important biological and metabolic functions that cannot be performed by researchers. Genomic and environmental factors at the basis of mutual host-microbiota interactions have been intensely investigated with metagenomic and metabolomic approaches in the last 5 years. This article will discuss recent advances in understanding the role of gut microbiota in the pathogenesis of obesity, insulin resistance (IR), and diabetes and their potential therapeutic applications.     

Protective effects of selenium-enriched peptides from Cardamine violifolia against high-fat diet induced obesity and its associated metabolic disorders in mice

Selenium-enriched peptides from Cardamine violifolia (CSP) have excellent antioxidant functions but little is known about their effects on obesity and associated metabolic disorders in mice fed with a high-fat diet (HFD). In this study, C57BL/6 mice were fed a HFD with or without CSP supplementation (CSPL: 26 μg Se per kg bw per d; CSPH: 104 μg per kg bw per d) for 10 weeks. The results showed that both CSPL and CSPH could ameliorate overweight gain, excess fat accumulation, serum lipid metabolism, and insulin resistance. The potential mechanism might be associated with the increase in thermogenesis, reduced oxidative stress, and inflammation, which regulated the gene expression in lipid and cholesterol metabolism. In addition, CSPL and CSPH also maintained the intestinal integrity and modulated the gut microbiota. Increased Blautia in CSP may be involved in the protective effect against obesity. Furthermore, a distinct increase in Lactobacillus was exclusively found in CSPH, suggesting that a more effective function of CSPH on metabolic disorders might be through the synergism of Blautia and Lactobacillus. Spearman''s correlation analysis revealed that these specific genera were significantly correlated with the metabolic improvements. Taken together, CSP supplementation prevented HFD-induced obesity and metabolic disorders, probably by ameliorating oxidative stress and inflammation, regulating metabolic genes, and modulating the gut microbiota compositions.

Selenium-enriched peptides from Cardamine violifolia (CSP) have excellent antioxidant functions but little is known about their effects on obesity and associated metabolic disorders in mice fed with a high-fat diet (HFD).     

Differences in gut microbiota between silkworms (Bombyx mori) reared on fresh mulberry (Morus alba var. multicaulis) leaves or an artificial diet

Artificial diets for silkworms have many potential applications and they are important in sericulture. However, the challenges of weak larvae and low silk protein synthesis efficiency in silkworms reared on artificial diets have not been resolved. Here, we used high-throughput sequencing to analyse the differences between the gut microbiota of 5th-instar larvae reared on mulberry leaves and larvae reared on an artificial diet. The results showed that at the phylum level, Cyanobacteria, Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria are the dominant bacteria in the intestines of silkworm larvae of all the strains. But the abundance of dominant bacteria in the gut microbiota differed between the silkworm strains that were reared on mulberry leaves, as well as between the silkworm strains that were reared on the artificial diet, while the gut microbiota diversity was lower in the silkworm strains that were reared on the artificial diet. Prediction of the functions of the gut microbiota in the hosts indicated that there was no significant difference between the silkworm strains that were reared on mulberry leaves, while there were significant differences between silkworm strains reared on the artificial diet. When the silkworm diet changed from mulberry leaves to the artificial diet, changes in gut microbiota in the silkworms affected host nutrient metabolism and immune resistance. These changes may be related to the adaptation of silkworms to their long evolutionary history of eating mulberry leaves.

Artificial diets for silkworms have many potential applications and they are important in sericulture.     

Anti-inflammatory activity of a water-soluble polysaccharide from the roots of purple sweet potato

In this study, a water-soluble polysaccharide was isolated from purple sweet potato roots. The in vitro and in vivo anti-inflammatory effects of the polysaccharide were evaluated by lipopolysaccharide (LPS)-induced inflammatory RAW264.7 macrophages and mice, respectively. The in vitro anti-inflammatory assay showed that the polysaccharide could effectively inhibit the overproduction of nitric oxide and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) while increasing the secretion of anti-inflammatory cytokine (IL-10). The in vivo anti-inflammatory assay revealed that mice administered with the polysaccharide showed higher IL-10, SOD, and T-AOC levels but lower TNF-α, IL-1β, IL-6 and MDA levels as compared to the LPS-treated model. Meanwhile, mice administered with the polysaccharide showed increased abundance of Lachnospiraceae, Lactobacillales and Parabacteroides but decreased amounts of Psychrobacter and Staphylococcus as compared to the LPS model group. Moreover, mice administered with polysaccharide showed enhanced production of short chain fatty acids by gut microbiota in the lipopolysaccharide-induced inflammatory mice. Our results suggested that the water-soluble polysaccharide from purple sweet potato roots could be utilized as a novel anti-inflammatory agent.

A water-soluble polysaccharide from purple sweet potato roots played anti-inflammatory roles by regulating inflammatory cytokines, gut microbiota and antioxidant defense system.     

Lower systemic inflammation is associated with gut firmicutes dominance and reduced liver injury in a novel ambulatory model of parenteral nutrition

BackgroundTotal Parenteral Nutrition (TPN) provides lifesaving nutritional support to patients unable to maintain regular enteral nutrition (EN). Unfortunately, cholestasis is a significant side effect affecting 20–40% of paediatric patients. While the aetiology of TPN-associated injury remains ill-defined, an altered enterohepatic circulation in the absence of gut luminal nutrient content during TPN results in major gut microbial clonal shifts, resulting in metabolic endotoxemia and systemic inflammation driving liver injury and cholestasis.HypothesisTo interrogate the role of gut microbiota, using our novel ambulatory TPN piglet model, we hypothesized that clonal reduction of bacteria in Firmicutes phylum (predominant in EN) and an increase in pathogenic Gram-negative bacteria during TPN correlates with an increase in serum lipopolysaccharide and systemic inflammatory cytokines, driving liver injury.MethodsUpon institutional approval, 16 animals were allocated to receive either TPN (n = 7) or EN only (n = 9). The TPN group was subdivided into a low systemic inflammation (TPN-LSI) and high systemic inflammation (TPN-HSI) based on the level of serum lipopolysaccharide. Culture-independent identification of faecal bacterial populations was determined by 16S rRNA.ResultsPiglets on TPN, in the TPN-HSI group, noted a loss of enterocyte protective Firmicutes bacteria and clonal proliferation of potent inflammatory and lipopolysaccharide containing pathogens: Fusobacterium, Bacteroidetes and Campylobacter compared to EN animals. Within the TPN group, the proportion of Firmicutes phylum correlated with lower portal lipopolysaccharide levels (r = −0.89). The TPN-LSI had a significantly lower level of serum bile acids compared to the TPN-HSI group (7.3 vs. 60.4 mg/dL; p = .018), increased day 14 weight (5.67 vs. 5.07 kg; p = .017) as well as a 13.7-fold decrease in serum conjugated bilirubin.ConclusionWe demonstrate a novel relationship between the gut microbiota and systemic inflammation in a TPN animal model. Pertinently, the degree of gut dysbiosis correlated with the severity of systemic inflammation. This study underscores the role of gut microbiota in driving liver injury mechanisms during TPN and supports a paradigm change in therapeutic targeting of the gut microbiota to mitigate TPN-related injury.

KEY MESSAGES

• This study identified a differential link between gut microbiota and inflammation—the higher the dysbiosis, the worse the systemic inflammatory markers.
• Higher levels of Firmicutes species correlated with reduced inflammation.

     

Association between metabolic profile and microbiomic changes in rats with functional dyspepsia

Functional dyspepsia (FD) is one of the most prevalent functional gastrointestinal disorders (FGIDs). Accumulated evidence has shown that FD is a metabolic disease that might relate to gut microbiota, but the relationship between microbiome and the host metabolic changes is still uncertain. To clarify the host–microbiota co-metabolism disorders related to FD, an integrated approach combining ¹H NMR-based metabolomics profiles, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and 16S rRNA gene sequencing was used to investigate the relationship among FD, metabolism of gut microbiota and the host. 34 differential urinary metabolites and 19 differential fecal metabolites, which affected the metabolism of energy, amino acids, nucleotides and short chain fatty acids (SCFAs), were found to have associated with FD. Based on the receiver operating characteristic (ROC) analysis, 10 biomarkers were screened out as diagnostic markers of FD. Meanwhile, the concentrations of Flintibacter, Parasutterella, Eubacterium and Bacteroides significantly increased in the FD group, whereas Eisenbergiella, Butyrivibrio, Intestinimonas, Saccharofermentans, Acetivibrio, Lachnoanaerobaculum and Herbinix significantly decreased. Furthermore, the above altered microbiota revealed a strong correlation with the intermediate products of the tricarboxylic acid (TCA) cycle, amino acids and SCFAs. In our study, it suggested that the energy metabolism was mainly disturbed in FD rats. Our findings also demonstrated that FD might be the result of gut microbiota and metabolism disorders, which was potentially valuable to enrich our understanding of the pathogenesis of FD.

Functional dyspepsia (FD) is one of the most prevalent functional gastrointestinal disorders (FGIDs). The aim of our study was to evaluate the effects of FD on the microbiota and its metabolic profiles in feces and urine.     

The in vitro fermentation of compound oral liquid by human colonic microbiota altered the abundance of probiotics and short-chain fatty acid production

Compound oral liquid (COL), made from functional herbal foods, has gained immense popularity in China for healthcare. However, the interaction between the nutrients in COL and gut microbiota is still unclear. In our study, the content of total flavonoids, polyphenols, and proteins was increased and the total sugar reduced by crushing raw ingredients to 10 mesh (COL-C). After 24 h incubation with supplemented COL by human gut microbiota, the results of 16S rRNA high-throughput sequencing revealed that Faecalibacterium, Collinsella, Bifidobacterium, Megamonas, Lactobacillus, Phascolarctobacterium, and Dialister were enriched by COL. In particular, the latter three genera were observed to be significantly enriched after incubation with COL-C. Meanwhile, the abundance of Dorea, Clostridium XIVa, and Escherichia/Shigella was inhibited by COL. Moreover, the increased levels of acetate, propionate, and butyrate in COL were jointly contributed by supplementary carbohydrates and the enrichment of short-chain fatty acid (SCFA)-producing bacteria. In summary, our results indicated that the optimized extraction facilitated the nutrients to be dissolved out and enhanced the potential prebiotic effects for promoting the abundance of probiotics, suggesting that the nutrients in COL-C might improve the microbial structure by strengthening the metabolism of beneficial bacteria and restricting the conditioned pathogens more efficiently.

The crushing pretreatment before extraction facilitated the nutrients to dissolve in compound oral liquid and enhanced the prebiotic effects for promoting the abundance of probiotics and short-chain fatty acid synthesis.     

An integrated metabolomics and 16S rRNA gene sequencing approach exploring the molecular pathways and potential targets behind the effects of Radix Scrophulariae

To assess the impact of the caecal microbiota on faecal metabolic phenotypes in the presence of Radix Scrophulariae (Chinese name: Xuanshen), an integrated approach involving 16S rRNA gene sequencing combined with ultrahigh-performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOF-MS)-based faecal metabolomics was applied to Radix Scrophulariae-treated rats. Interestingly, Radix Scrophulariae led to significant gut microbiota changes at the phylum and genus levels in treated rats compared to control rats. Additionally, distinct changes in faecal metabolites, including linoleic acid (LA), guanosine, inosine, hypoxanthine, xanthine, 4-hydroxycinnamic acid, cholic acid, N-acetyl-d-glucosamine, l-urobilinogen and uridine, were observed in Radix Scrophulariae-treated rats. Of these, seven metabolites were up-regulated, and the remaining three metabolites were down-regulated. Moreover, there were substantial associations between altered levels of gut microbiota genera and discrepant levels of faecal metabolites, particularly for compounds involved in LA and purine metabolism. These results demonstrated that the gut microbiota is altered in association with faecal metabolism following treatment with Radix Scrophulariae. Our findings suggest that further application of this 16S rRNA gene sequencing and UHPLC/TOF-MS-based metabolomics approach will facilitate the assessment of the pharmacological action of Radix Scrophulariae and thus expand the scope of this herb.

An integrated approach to assess impact of caecal microbiota on faecal metabolic phenotypes in the presence of Radix Scrophulariae involving 16S rRNA gene sequencing combined with UHPLC/TOF-MS-based faecal metabolomics was applied to Radix Scrophulariae-treated rats.     

Is there a role for gut microbiota in type 1 diabetes pathogenesis?

Type 1 diabetes mellitus (T1D) is an autoimmune disease characterized by insufficient insulin production due to the destruction of insulin secreting β-cells in the Langerhans islets. A variety of factors, including chemicals, viruses, commensal bacteria and diet have been proposed to contribute to the risk of developing the disorder. In the last years, gut microbiota has been proposed as a main factor in T1D pathogenesis. Several alterations of gut microbiota composition were described both in animal model and in humans. The decrease of Firmicutes/Bacteroides ratio was the most frequent pattern described, in particular, in human studies. Furthermore, Bacteroides, Clostridium cluster XIVa, Lactobacillus, Bifidobacterium, and Prevotella relative abundances were different in healthy and affected subjects. Dysbiosis would seem to increase intestinal permeability and thus promote the development of a pro-inflammatory niche that stimulates β-cell autoimmunity in predisposed subjects. Preliminary studies on animal models were realized to investigate the role of gut microbiota modulation as therapy or prevention approach in predisposed animals: promising and stimulating results have been reported.

• Key message

• Dietary antigens and microbiota-derived products might act as triggers of T1D by causing a pro-inflammatory and metabolic dysfunctional environment.

     

Impact of purple sweet potato (Ipomoea batatas L.) polysaccharides on the fecal metabolome in a murine colitis model

Purple sweet potato polysaccharides (PSPP) play an important role in regulating the gut microbiota, modulating intestinal immunity and ameliorating colonic inflammation. In this study, the impact of two PSPPs (PSWP-I and PSAP-I) on the metabolomic profiling of feces from dextran sulfate sodium (DSS)-induced colitis mice was evaluated by ultra-high performance liquid chromatography coupled with triple time-of-flight tandem mass spectrometry (UPLC-Triple-TOF-MS/MS). Results indicated that there were twenty-five metabolites with significant changes and four remarkable metabolic pathways, i.e., cutin, suberine and wax biosynthesis, biosynthesis of unsaturated fatty acids, fatty acid biosynthesis, and steroid hormone biosynthesis. Two key biomarkers of oleic acid and 17-hydroxyprogesterone were screened that responded to PSPPs in colitis mice. The identified metabolites were correlated with the amelioration of intestinal immune function and the modulation of the gut microbiota. Nine pro-inflammatory and eight anti-inflammatory compounds responded to PSPPs, which were related to Bacteroides, norank_f__Clostridiales_vadinBB60_group, unclassified_o__Bacteroidales, Rikenella and Lachnospiraceae_UCG-001. Moreover, PSWP-I and PSAP-I had different regulating effects on intestinal metabolites. Our results revealed a possible metabolomic mechanism of PSPPs to regulate intestinal inflammation function.

Purple sweet potato polysaccharides (PSPP) play an important role in regulating the gut microbiota, modulating intestinal immunity and ameliorating colonic inflammation.     

Mitochondrial dysfunction in inflammatory bowel disease alters intestinal epithelial metabolism of hepatic acylcarnitines

As the interface between the gut microbiota and the mucosal immune system, there has been great interest in the maintenance of colonic epithelial integrity through mitochondrial oxidation of butyrate, a short-chain fatty acid produced by the gut microbiota. Herein, we showed that the intestinal epithelium could also oxidize long-chain fatty acids, and that luminally delivered acylcarnitines in bile could be consumed via apical absorption by the intestinal epithelium, resulting in mitochondrial oxidation. Finally, intestinal inflammation led to mitochondrial dysfunction in the apical domain of the surface epithelium that may reduce the consumption of fatty acids, contributing to higher concentrations of fecal acylcarnitines in murine Citrobacter rodentium–induced colitis and human inflammatory bowel disease. These results emphasized the importance of both the gut microbiota and the liver in the delivery of energy substrates for mitochondrial metabolism by the intestinal epithelium.     

The fecal metabolome is associated with gestational diabetes mellitus

Dysbiosis of gut microbiota has been linked to gestational diabetes mellitus (GDM), and grows as a resource for GDM biomarkers. However, the contributions of gut microbiota to GDM remain incompletely understood. Metabolites are key messengers in the interactions between gut microbiota and the host. Metabolomics is emerging as an essential tool in exploring the contributions of gut microbiota to diseases. In this study, we performed ¹H-NMR based metabolomics on the feces of 62 pregnant women, including 31 women with GDM, and 31 women as the non-diabetes (NDM) control. Using Principle Component Analysis (PCA) and Orthogonal Projection to Latent Structures Discrimination Analysis (OPLS-DA), we observed clear cluster separation of the fecal metabolome between women with GDM and the NDM control. We further applied several feature selection methods to find five fecal metabolites contributing to the cluster separation of the fecal metabolome. These five metabolites, namely dibutyl decanedioate, N-acetylgalactosamine-4-sulphate, homocysteine, l-malic acid, and butanone, were significantly correlated with the clinical indices of GDM. Metabolite enrichment and pathway analysis on the five metabolites suggested that the fecal citrate cycle and sulfur metabolism were correlated with GDM. The results of this study demonstrated that disorders in the fecal metabolome are associated with GDM.

Fecal metabolome could separate women with GDM from the non-diabetic control.     

Effects of dopexamine on the intestinal microvascular blood flow and leucocyte activation in a sepsis model in rats

Introduction

Dopexamine may be a therapeutic option to improve hepatosplanchnic perfusion in sepsis. To investigate this possibility, we administered dopexamine in an experimental sepsis model in rats.

Methods

This prospective, randomized, controlled laboratory study was conducted in 42 Wistar rats. The animals were divided into three groups. Group 1 served as the control group (CON group). The animals in both groups 2 (LPS group) and 3 (DPX group) received an endotoxin (lipopolysaccharide from Escherichia coli – LPS) infusion (20 mg/kg for 15 minutes). DPX group additionally received dopexamine (0.5 μg/kg per minute over four hours). One half of the animals in each group underwent studies of intestinal microvascular blood flow (IMBF) using laser Doppler fluxmetry. In the other half an intravital microscopic evaluation of leucocyte-endothelial cell interaction in intestinal microcirculation was conducted. Functional capillary density (FCD) in the intestinal mucosa and in the circular as well as longitudinal muscle layer was estimated.

Results

One hour after endotoxin challenge, IMBF decreased significantly in LPS group to 51% compared with baseline (P < 0.05). In DPX group (endotoxin plus dopexamine) we found IMBF values significantly higher than those in LPS group (approximately at the level of controls). The impaired FCD following endotoxin challenge was improved by dopexamine in the longitudinal muscle layer (+33% in DPX group versus LPS group; P < 0.05) and in the circular muscle layer (+48% in DPX group versus LPS group; P < 0.05). In DPX group, dopexamine administration reduced the number of firmly adherent leucocytes (-31% versus LPS group; P < 0.05). Plasma levels of tumour necrosis factor-α were reduced by dopexamine infusion (LPS group: 3637 ± 553 pg/ml; DPX group: 1933 ± 201 pg/ml) one hour after endotoxin challenge.

Conclusion

Dopexamine administration improved IMBF and FCD (markers of intestinal microcirculation) and reduced leucocyte activation (a marker of inflammation) in experimental sepsis.     

Metabonomic strategy for the detection of metabolic effects of probiotics combined with prebiotic supplementation in weaned rats

The purpose of this study is to investigate the effects of probiotics combined with prebiotics (PP) supplementation on weaned rat metabolism. A metabonomic strategy employing ¹H-NMR spectroscopy and multivariate data analysis was used to examine weaned rat biological responses to PP supplementation. Male Sprague-Dawley rats (post-natal day 21, PD 21) received probiotics (Lactobacillus acidophilus NCFM (L-NCFM) and Bifidobacterium lactis Bi-07 (B-LBi07), 1 : 1, 1.0 × 10¹¹ cfu kg⁻¹) and prebiotics (Lycium barbarum polysaccharides (LBP), Poria cocos polysaccharides (PCPs) and Lentinan, 1 : 1 : 1, 24 g kg⁻¹) via intragastric administration for 28 consecutive days. Urine and feces were collected for analysis. Significant topographical metabolic variations were present in urine and feces. Urinary metabolites upregulated by PP treatment included alanine, N-acetylglycine, glutamine, dimethylamine, phosphorylcholine, ethylene glycol, mannitol, phenylacetylglycine and glycoate, which were related to alanine, aspartate and glutamate metabolism, and choline metabolism. Feces-derived metabolites, including caproate, valerate, butyrate, propionate, lactate, acetate, succinate, methanol, threonine and methionine, were significantly increased, which were related to short-chain fatty acid (SCFA) metabolism and TCA cycle metabolism. These results indicate that dietary PP supplementation can regulate common systemic metabolic processes, including energy metabolism, amino acid metabolism, lipid metabolism, nucleic acid metabolism, and gut microbiota-related metabolism. This study also illuminates the vital role of PP supplementation in regulating the metabolism of weaned rats.

Dietary probiotic supplementation is beneficial to the growth of weaned rats by regulating the metabolism.     

Consumption of post-fermented Jing-Wei Fuzhuan brick tea alleviates liver dysfunction and intestinal microbiota dysbiosis in high fructose diet-fed mice

Emerging evidence supports the health-promoting ability of a special microbial-fermented Fuzhuan brick tea. Epigallocatechin gallate was identified as a dominant flavonoid of Fuzhuan tea aqueous extract (FTE). Mice were treated with 30% high fructose (HF) water feeding alone or in combination with administration of FTE at 400 mg per kg bw for 13 weeks. FTE caused strong inhibition against the elevation of liver weight, serum enzymatic (aspartate aminotransferase, aspartate aminotransferase and alkaline phosphatase) activities and hepatic inflammatory cytokines (interleukin-1, interleukin-6, tumor necrosis factor-α and tumor necrosis factor-β) formation, as well as dyslipidemia (total cholesterol, total triglyceride, low-density lipoprotein-cholesterol and high-density lipoprotein-cholesterol) in HF-fed mice (p < 0.05). Hepatic malonaldehyde formation was lowered, while superoxide dismutase and glutathione peroxidase activities were enhanced by FTE treatment, relative to HF-fed mice (p < 0.05), and histopathological evaluation confirmed the protection. As revealed by 16S rDNA gene sequencing, FTE notably increased abundance of Bacteroidetes and Lactobacillus, but reduced population of Firmicutes, Proteobacteria and Tenericutes in HF feeding mice. These findings suggest that FTE exerts a hepatoprotective effect by modifying hepatic oxidative stress, inflammatory response and gut microbiota dysfunction.

Fuzhuan brick tea can improve non-alcoholic fatty liver disease (NAFLD) and intestinal microbiota imbalance induced by a high fructose diet (HFD) intake in mice.