Fecal Water Content Research Articles

Water Extract of Ecklonia cava Protects against Fine Dust (PM2.5)-Induced Health Damage by Regulating Gut Health.

To confirm the therapeutic effect of the water extract from Ecklonia cava (WEE) against PM2.5 induced systemic health damage, we evaluated gut health with a focus on the microbiota and metabolites. Systemic damage in mice was induced through PM2.5 exposure for 12 weeks in a whole-body chamber. After exposure for 12 weeks, body weight and food intake decreased, and WEE at 200 mg/kg body weight (mpk) alleviated these metabolic efficiency changes. In addition, PM2.5 induced changes in the length of the colon and fecal water content. The administration of the WEE at 200 mpk oral dose effectively reduced changes in the colon caused by PM2.5 exposure. We also attempted to confirm whether the effect of the WEE is mediated via regulation of the microbiota-gut-brain axis in mice with PM2.5 induced systemic damage. We examined changes in the fecal microbiota and gut metabolites such as short-chain fatty acids (SCFAs) and kynurenine metabolites. In the PM2.5 exposed group, a decrease in the abundance of Lactobacillus (Family: Lactobacillaceae) and an increase in the abundance of Alistipes (Family: Rikenellaceae) were observed, and the administration of the WEE showed a beneficial effect on the gut microbiota. In addition, the WEE effectively increased the levels of SCFAs (acetate, propionate, and butyrate). Furthermore, kynurenic acid (KYNA), which is a critical neuroprotective metabolite in the gut-brain axis, was increased by the administration of the WEE. Our findings suggest that the WEE could be used as a potential therapeutic against PM2.5 induced health damage by regulating gut function.

Momordica charantia polysaccharides alleviate diarrhea-predominant irritable bowel syndrome by regulating intestinal inflammation and barrier via NF-κB pathway.

Momordica charantia exerts anti-inflammatory effect against ulcerative colitis. Momordica charantia polysaccharides (MCPs) attenuate gastritis through inhibition of ethanol-induced inflammatory response. The role of MCPs in diarrhea-predominant irritable bowel syndrome (IBS-D) is investigated. Chemical stimulation followed by acute and chronic pressure stimulation was used to establish rats model with IBS-D. The model rats were then administrated with MCPs. Defecation frequency, fecal water content and abdominal withdrawal reflex (AWR) score were then recorded. Pathologic changes in the colonic tissues were evaluated by hematoxylin and eosin staining. Inflammation was detected by ELISA and qRT-PCR, and immunohistochemistry was used to assess intestinal mucosal permeability. First, IBS-D of mice wasIBS-D ratsmice exhibited many abnormal clinical manifestations, including increased frequency of defecation, fecal water content, and abdominal withdrawal reflex (AWR) score. Second, the mice were administrated with MCPs, which reduced frequency of defecation, fecal water content, and AWR score, and 100-mg/kg MCPs indicated therapeutic effect on IBS-D mice equivalent to rifaximin. Moreover, MCPs also ameliorated pathologic changes in the colonic tissues of IBS-D mice. Third, inflammatory response in IBS-D mice was also suppressed by MCPs through up-regulation of Interleukin (IL)-10, and down-regulation of tumor necrosis factor-α (TNF-α), Interleukin(IL)-1β, and IL-6. MCPs enhanced levels of occludin (OCLN) and zona occludens protein-1 (ZO-1) in IBS-D mice to improve intestinal mucosal permeability. Finally, phosphorylation of p65 in IBS-D mice was reduced by MCP treatment. MCPs ameliorated intestinal permeability and repressed intestinal inflammation to attenuate IBS-D by inactivating nuclear factor kappa B (NF-κB) signaling.

Bifidobacterium Treated by Electrostatic Spray Drying Relieved Constipation by Changing the Relative Abundance of Bacteria Associated With Gastrointestinal Regulatory Peptides.

In this study, three different microencapsulation methods were used to embed Bifidobacterium to explore the alleviating effects of embedding methods on constipated mice. By measuring the defecation-related parameters, it was found that the Bifidobacteria treated by electrostatic spray drying had the best ability to relieved constipation. Furthermore, by detecting constipation-related gastrointestinal regulatory peptides, inflammatory factors, intestinal microbiota, and SCFAs, it was discovered that Bifidobacteria treated by electrostatic spray drying changed the composition of intestinal microbiota, especially the relative abundance of bacteria that were positively correlated with AQP3, but negatively correlated with ET-1 and SS, then increased the level of AQP3 in the intestine, and finally relieved constipation by increasing the fecal water content and small intestinal propulsion rate. In conclusion, the electrostatic spray drying method was superior to the other two methods in maintaining the activity of Bifidobacteria and relieved constipation by changing the relative abundance of bacteria that were correlated with gastrointestinal regulatory peptides and increasing the content of fecal water and small intestinal propulsion rate.

Effects of Maren Pills on the Intestinal Microflora and Short-Chain Fatty Acid Profile in Drug-Induced Slow Transit Constipation Model Rats.

Background: Slow transit constipation (STC) is becoming a common and frequently occurring disease in today’s society, and it is necessary to explore the safe and effective treatment of STC. Method: Our study aimed to investigate whether the laxative effect of Maren pills (MRW) is associated with the regulation of intestinal microflora and intestinal metabolism in the colon. Loperamide hydrochloride-induced STC rats received MRW intragastrically for two consecutive weeks to evaluate the laxative effect of MRW involving the regulation of intestinal microflora, intestinal metabolism, and 5-HT signaling pathway. Intestinal microflora was detected by 16s rDNA sequencing, intestinal metabolism of short-chain fatty acids (SCFAs) was detected by HPLC, and the 5-HT signaling pathway was detected by WB, ELISA, immunofluorescence, and immunohistochemical analysis. Results: Our results revealed that the treatments with MRW increased not only the body weight, 24-h fecal number, 24-h wet fecal weight, 24-h dry fecal weight, fecal water content, and the intestinal propulsion rate but also the colonic goblet cell number, colonic Muc-2 protein expression, and colonic mucus layer thickness in the STC model rats. Moreover, MRW activated the 5-HT pathway by increasing the levels of 5-HT, 5-HIAA, 5-HT4R, CFTR, cAMP, and PKA in the colon tissue of STC rats. The 16S rDNA sequencing results showed that MRW improved the colonic microflora structure in colonic contents of STC rats, mainly by increasing Lactobacillus and decreasing Prevotella. Finally, we found that MRW regulated the SCFA metabolism in the colonic contents of the STC rats, mainly by increasing the contents of acetic acid, propionic acid, and butyric acid; the relative abundance of Lactobacillus was positively correlated with either contents of acetic acid, propionic acid, and butyric acid, and the relative abundance of Clostridium was negatively correlated. Conclusion: Our study further showed that MRW could improve constipation in STC rats, and the mechanism may be by regulating the intestinal microflora structure and improving the metabolism of SCFAs.

Metformin Inhibits Na+/H+ Exchanger NHE3 Resulting in Intestinal Water Loss.

Glycemic control is the key to the management of type 2 diabetes. Metformin is an effective, widely used drug for controlling plasma glucose levels in diabetes, but it is often the culprit of gastrointestinal adverse effects such as abdominal pain, nausea, indigestion, vomiting, and diarrhea. Diarrhea is a complex disease and altered intestinal transport of electrolytes and fluid is a common cause of diarrhea. Na+/H+ exchanger 3 (NHE3, SLC9A3) is the major Na+ absorptive mechanism in the intestine and our previous study has demonstrated that decreased NHE3 contributes to diarrhea associated with type 1 diabetes. The goal of this study is to investigate whether metformin regulates NHE3 and inhibition of NHE3 contributes to metformin-induced diarrhea. We first determined whether metformin alters intestinal water loss, the hallmark of diarrhea, in type 2 diabetic db/db mice. We found that metformin decreased intestinal water absorption mediated by NHE3. Metformin increased fecal water content although mice did not develop watery diarrhea. To determine the mechanism of metformin-mediated regulation of NHE3, we used intestinal epithelial cells. Metformin inhibited NHE3 activity and the effect of metformin on NHE3 was mimicked by a 5′-AMP-activated protein kinase (AMPK) activator and blocked by pharmacological inhibition of AMPK. Metformin increased phosphorylation and ubiquitination of NHE3, resulting in retrieval of NHE3 from the plasma membrane. Previous studies have demonstrated the role of neural precursor cell expressed, developmentally down-regulated 4-2 (Nedd4-2) in regulation of human NHE3. Silencing of Nedd4-2 mitigated NHE3 inhibition and ubiquitination by metformin. Our findings suggest that metformin-induced diarrhea in type 2 diabetes is in part caused by reduced Na+ and water absorption that is associated with NHE3 inhibition, probably by AMPK.

Preliminary study on time-dependent changes of intestinal tract and brain-gut axis in mice model of Parkinson's disease induced by paraquat

Objective: To observe the intestinal time-dependent changes in Parkinson's disease (PD) mouse model constructed by intraperitoneal injection of paraquat (PQ) and to establish the brain-gut axis connection initially. Methods: In October 2019, 48 mice were randomly divided into treated group and control groups: treated 4-week (P-4) group, treated 6-week (P-6) group, treated 8-week (P-8) group, control 4-week (C-4) group, control 6-week (C-6) group, and control 8-week (C-8) group. The treated group was injected with 15 mg/kg PQ solution and the control group was injected with 0.9% saline (0.2 ml/20 g) by intraperitoneal injection twice a week. After the initial state (0 weeks) and the treatment at the end of 4, 6 and 8 weeks, the mood changes and motor functions of mice were assessed by neurobehavioral tests (open field test, pole climbing test, tail suspension test and elevated plus maze test) . And the number of fecal pellets for 1 h and water content were calculated to assess the functional status of the gastrointestinal tract. Western blotting experiments were performed to detect the expression levels of α-synuclein (α-syn) and tyrosine hydroxylase (TH) in the nigrostriatal region of the mouse brain, the tight junction markers zonula occludens-1 (ZO-1) and Occludin, the inflammatory markers of integrin αM subunit (CD11b) , inducible nitric oxide synthase (iNOS) , high mobility group box 1 (HMGB1) , interleukin-1β (IL-1β) , and the neuronal markers βⅢ-tubulin and α-syn protein in the colon.Immunohistochemical staining was performed to detect the expression levels of colonic tight junction proteins ZO-1 and Occludin. Immunofluorescence staining was performed to detect the expression levels of TH in the substantia nigra region of the midbrain, and the co-localization of colonic intestine neuronal marker (βⅢ-tubulin) and Ser129 α-syn in the colonic. Results: Compared with the initial state (0 weeks) and C-8 group, mice in the P-8 group had significantly higher pole climbing test scores and resting time, and significantly lower total active distance, mean active speed, percentage of open arm entry and 1 h fecal instances (P<0.05) . After poisoning, the 1 h fecal water content of model mice first increased and then decreased, the P-4 and P-6 groups were significantly higher than the simultaneous point control group, and the P-8 groups were significantly lower than the initial state (P<0.05) . Compared with control, P-4 and P-6 groups, the expression levels of ZO-1 and Occludin in the P-8 group were significantly decreased (P<0.05) . Compared with control group, the expression levels of CD11b and IL-1β in the P-4 group were significantly increased (P<0.05) . Compared with control and P-4 group, the expression levels of CD11b, iNOS, HMGB1 and IL-1β in the P-6 and P-8 groups were significantly increased (P<0.05) . Compared with the control and P-4 groups, the expression levels of βⅢ-tubulin in the colon of mice in the P-8 group were significantly decreased, and the expression levels of α-syn and Ser129 α-syn were significantly increased (P<0.05) . The expression level of Ser129 α-syn in the colon of model mice was negatively correlated with the expression level of βⅢ-tubulin (r(s)=-0.9149, 95%CI: -0.9771--0.7085, P<0.001) . Ser129 α-syn and βⅢ-tubulin co-localization in the colonic intermuscular plexus region increased gradually with the time of exposure. Compared with the control, P-4 and P-6 groups, the expression level of TH in the nigrostriatal region of the brain was significantly decreased, and the expression levels of α-syn and Ser129 α-syn were significantly increased in the P-8 group (P<0.05) . Correlation analysis showed that the relative expression level of Ser129 α-syn in the nigrostriatal region of the brain was negatively correlated with the expression level of TH in the model mice (r(s)=-0.9716, 95% CI: -0.9925--0.8953, P<0.001) . Conclusion: The PD mouse model is successfully established by PQ, and the intestinal function of the model mice is reduced in a time-dependent manner. And on this basis, it is preliminary determined that the abnormal aggregation of α-syn may be an important substance connecting the brain-gut axis.

Assessing animal welfare: a triangulation of preference, judgement bias and other candidate welfare indicators

To assess the welfare of captive animals, validated measures, so-called ‘welfare indicators’, are required. We used a triangulation approach to investigate the extent to which different measures converged to provide corroborating evidence of welfare. Laying hens were exposed to living conditions designed to be generally preferred (GP) or generally nonpreferred (GNP), using previous studies of chickens’ majority preferences for resources and environments. The hens were also tested at the end of the study to identify their individual preferences for these living conditions, assigned to groups that showed an individual preference, or nonpreference, for their own experimental housing, regardless of whether it was generally preferred or not (IP and INP). Both GP and IP birds showed more ground-foraging behaviour, and lower pulse rates during handling, than GNP and INP birds. Individual preference was associated with more optimistic-like judgement biases when birds were tested after 6 weeks of exposure to these living conditions, but not after 24 weeks. Serum blood glucose levels were also lower in hens showing individual preferences for their experimental living conditions. General preferences were associated with a number of measures, including higher rates of ground-foraging behaviour and lower faecal water content (after both 6 and 24 weeks), lower pulse rate during handling and greater tibia strength and stiffness post mortem. There were no associations between judgement bias and other candidate welfare indicators, but it is not clear whether this represents evidence of absence or merely absence of evidence. Overall, the different approaches did not converge to identify a precise state of animal welfare, although some measures (preference, stress indicators: pulse rate, faecal water and blood glucose, foraging behaviour) were aligned consistently across timescales. We conclude that further work is needed to establish which alternative measures of affective state might be more appropriate indicators of animal welfare.

Modulation of gut microbiota during alleviation of antibiotic-associated diarrhea with Zingiberis Rhizoma

This study was aimed to explore the effect of Zingiberis Rhizoma extract on rats with antibiotic-associated diarrhea(AAD), and reveal the modulation of gut microbiota during alleviation of AAD. AAD rat model was successfully established by exposing rats to appropriate antibiotic mixed solution. Peficon(70 mg·kg~(-1)·d~(-1)) was used as positive control, then rats were treated with 200 mg·kg~(-1)·d~(-1) and 400 mg·kg~(-1)·d~(-1) of Zingiberis Rhizoma extract for low and high dosage groups of Zingiberis Rhizoma extract, respectively. The weight changes of the rats were observed, and the degree of diarrhea were evaluated by fecal score, 120 min fecal weight and fecal water content. Colon tissues for histopathological examination were stained with hematoxylin and eosin(HE), and 16 S rRNA sequencing analysis of gut microbiota was performed. The results showed that compared with the model group, the degree of diarrhea, indicated by fecal water content, fecal score, and 120 min fecal weight of positive control group, Zingiberis Rhizoma low-dose group and Zingiberis Rhizoma high-dose group were significantly ameliorated. And the treatment of Zingiberis Rhizoma could significantly improve the pathological condition of colon tissue in AAD rats, especially the high dose of Zingiberis Rhizoma. In addition, 16 S rRNA sequencing analysis of gut microbiota showed that the diversity and abundance of gut microbiota were significantly improved and the reco-very of gut microbiota was accelerated after given high-dose of Zingiberis Rhizoma, while no significant changes of alterations were observed after given Pefikon. Of note, compared with the pefikon group, the abundance and diversity of gut microbiota in Zingi-beris Rhizoma high-dose group were significantly elevated. At the phylum level, the abundance of Firmicutes in AAD rats increased and the abundance of Proteobacteria was decreased after the Zingiberis Rhizoma intervention. At the genus level, the abundance of Bacillus spp., Lachnoclostridium and Escherichia coli-Shigella were decreased, and the abundance of Lactobacillus spp., Trichophyton spp., and Trichophyton spp., etc., were increased. While compared with the AAD model group, there was no significant difference of gut microbiota after given Peficon. The results showed that Zingiberis Rhizoma exerted beneficial health effects against AAD, and positively affected the microbial environment in the gut of rats with AAD.

SAHA Alleviates Diarrhea-Predominant Irritable Bowel Syndrome Through Regulation of the p-STAT3/SERT/5-HT Signaling Pathway.

ObjectiveIrritable bowel syndrome (IBS) is characterized by abdominal pain, bloating, and stool irregularity. However, its pathophysiological mechanisms, which trigger intestinal motility disorders and diarrhea leading to diarrhea-predominant IBS (D-IBS), remain largely unknown.MethodsIn the present study, we established a D-IBS rat model by mother–infant separation combined with restraint stress. Then we exposed the modelled rats to suberoylanilide hydroxamic acid (SAHA) treatment, followed by determination of their visceral sensitivity. Toluidine blue staining served to reveal the effects of SAHA treatment on mast cells of D-IBS model rats. Then we measured the expression of serotonin (5-hydroxytryptamine; 5-HT) and its receptors by ELISA.ResultsConstruction of short hairpin RNA (sh)-serotonin transporter (SERT) lentivirus vectors verified the regulation of the 5-HT signaling pathway by phosphorylated (p)-STAT/SERT. SAHA treatment of D-IBS model rats reduced the fecal water content, electromyography integral change rate, abdominal withdrawal reflex score, and number of mast cells, as well as the expression of 5-HT type 3A (5-HT3AR), 3B receptor (5-HT3BR), and 4 receptor (5-HT4R) receptors. The treatment also elevated the expression of signal transducer and activator for transcription 3 (STAT3) and SERT. Activation of p-STAT3 may reverse the inhibitory effect of SAHA on the elevated visceral sensitivity of D-IBS model rats. Moreover, SAHA promoted the transcription of SERT through repression of the p-STAT3/5-HT signaling, thereby inhibiting the visceral sensitivity of D-IBS model rats.ConclusionThis study highlights that SAHA treatment can alleviate D-IBS through regulation of the p-STAT3/SERT/5-HT signaling pathway.

Ecology of the free-living stages of Trichostrongylid parasites of sheep

The prevalence of gastrointestinal nematodes (GINs) is dictated by environmental conditions which influence the survival, development, and migration rates of the free-living stages. While the faecal pellet is the most important micro-environment for GINs, limited data on the impact of temperature and faecal water content (FWC) on their survival, development, and migration limits accurate prediction of nematode pasture infectivity. This study investigated the impact of temperature and FWC on the dynamics of the free-living stages of T. vitrinus, T. colubriformis and T. circumcincta by incubating faecal samples from sheep harbouring mono-specific infection of the GINs in temperature-controlled incubators at constant temperatures ranging from 0° to 40°C and nominal target FWCs ranging from 0% to 80% for 8 days. The different life cycle stages (eggs, pre-infective larvae, intra-pellet infective larvae and extra-pellet infective larvae) were enumerated each day using a modified McMaster technique. Multiple linear regression analysis was used to assess the impact of environmental effects on the recovery of each life cycle stage. A one-way ANOVA was used to compare the life cycle stage tallies between species. Survival and development of each life cycle stage were elucidated by fitting the obtained experimental data to an existing mathematical model which described the dynamics of the free-living stages of the GIN lifecycle. Development of eggs to infective larvae were only observed at target temperatures of 20 and 30ºC (and some development at 40 °C for T. colubriformis). This study predicted development of a minimum of 1% of eggs to intra-pellet infective larvae at temperature ranges of 9–39, 10–39 and 10–38ºC with optimum development occurring at 23, 23 and 20ºC for T. vitrinus, T. circumcincta and T. colubriformis, respectively. Development of eggs to intra-pellet infective larvae was observed at nominal FWC targets of 20% and above where increasing FWC up to 60% resulted in increased tallies of intra-pellet infective larvae. The model predicted that development of eggs to intra-pellet infective larvae required a minimum FWC of 13%, 16% and 17% for T. circumcincta, T. colubriformis and T. vitrinus respectively with higher FWC requirements for migration out of the faecal pellets. Development of T. circumcincta eggs to infective larvae was slower than the other species with only 33% of the T. circumcincta eggs predicted to develop to infective larvae at its optimum temperature and FWC as compared to 47% for T. vitrinus and 56% for T. colubriformis.

Mucosal Serotonin Reuptake Transporter Expression in Irritable Bowel Syndrome Is Modulated by Gut Microbiota Via Mast Cell–Prostaglandin E2

Increased colonic serotonin (5-HT) level and decreased serotonin reuptake transporter (SERT) expression in irritable bowel syndrome (IBS) may contribute to diarrhea and visceral hypersensitivity. We investigated whether mucosal SERT is modulated by gut microbiota via a mast cell-prostaglandin E2 (PGE2) pathway. C57Bl/6 mice received intracolonic infusion of fecal supernatant (FS) from healthy controls or patients with diarrhea-predominant irritable bowel syndrome (IBS-D). The role of mast cells was studied in mast cell-deficient mice. Colonic organoids and/or mast cells were used for invitro experiments. SERT expression was measured by quantitative polymerase chain reaction and Western blot. Visceromotor responses tocolorectal distension and colonic transit were assessed. Intracolonic infusion of IBS-D FS in mice caused an increase in mucosal 5-HT compared with healthy control FS, accompanied by ∼50% reduction in SERT expression. Mast cell stabilizers, cyclooxygenase-2 inhibitors, and PGE2 receptor antagonist prevented SERT downregulation. Intracolonic infusion of IBS-D FS failed to reduce SERT expression in mast cell-deficient (W/Wv) mice. This response was restored by mast cell reconstitution. The downregulation of SERT expression evoked by IBS FS was prevented by lipopolysaccharide (LPS) antagonist LPS from Rhodobacter sphaeroides and a bacterial trypsin inhibitor. Invitro LPS treatment caused increased cyclooxygenase-2 expression and PGE2 release from cultured mouse mast cells. Intracolonic infusion of IBS-D FS in mice reduced colonic transit, increased fecal water content, and increased visceromotor responses to colorectal distension. Ondansetron prevented these changes. Fecal LPS acting in concert with trypsin in patients with IBS-D stimulates mucosal mast cells to release PGE2, which downregulates mucosal SERT, resulting in increased mucosal 5-HT. This may contribute to diarrhea and abdominal pain common in IBS.

Effect of traditional chinese medicine (TCM) and its fermentation using Lactobacillus plantarum on ceftriaxone sodium-induced dysbacteriotic diarrhea in mice

BackgroundBuzhongyiqi decoction (BD), Sijunzi decoction (SD), and Shenlingbaizhu decoction (SHD) have been extensively used clinically for the treatment of diseases caused by spleen-Qi deficiency and microbial fermentation has historically been utilized in traditional Chinese medicine (TCM). This study aimed to investigate the mitigative effect of TCM and fermented TCM (FTCM) with Lactobacillus plantarum (LP) on antibiotic-associated diarrhea, and to select an optimal formula and then identify its compounds.MethodsDysbacteriosis in mice was induced by ceftriaxone sodium (CS). The mice were then treated with LP, BD, SD, SHD, fermented BD, fermented SD (FSD), and fermented SHD. Diarrhea indexes, the abundances of gut bacteria, intestinal morphometrics, and mRNA expressions of genes related to intestinal barrier function were assessed. Then, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) were employed to identify and relatively quantify the compounds in the selected decoctions.ResultsCS significantly increased the fecal output weight, the total number of fecal output, and fecal water content, indicating the occurrence of diarrhea. Bacterial culture tests showed that the above symptoms were accompanied by the disruption of specific intestinal flora. TCM, LP, and FTCM alleviated the diarrhea index and recovered the intestinal microbiota. FTCM showed more advantageous than TCM or LP alone. The mRNA expressions of aquaporins (AQPs) and tight junctions (TJs) decreased by CS were enhanced by TCM, LP, and FTCM. In addition, through UHPLC-Q-TOF/MS, (S)-(-)-2-hydroxyisocaproic acid, L-methionine, 4-guanidinobutyric acid (4GBA), and phenyllactate (PLA) in SD and FSD were identified and relatively quantified.ConclusionsTCM, LP, and TCM fermented with LP alleviated CS-induced diarrhea symptoms, and improved the intestinal flora and barrier function. Four compounds including (S)-(-)-2-hydroxyisocaproic acid, L-methionine, 4GBA, and PLA in FSD, which were identified by UHPLC-Q-TOF/MS, might function in modulating intestinal flora and improving villi structure.

Targeting intestinal flora and its metabolism to explore the laxative effects of rhubarb.

Rhubarb, a traditional herb, has been used in clinical practice for hundreds of years to cure constipation, but its mechanism is still not clear enough. Currently, growing evidence suggests that intestinal flora might be a potential target for the treatment of constipation. Thus, the aim of this study was to clarify the laxative effect of rhubarb via systematically analyzing the metagenome and metabolome of the gut microbiota. In this study, the laxative effects of rhubarb were investigated by loperamide-induced constipation in rats. The gut microbiota was determined by high-throughput sequencing of 16S rRNA gene. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used for fecal metabolomics analysis. The data showed that rhubarb could significantly shorten gastrointestinal transit time, increase fecal water content and defecation frequency, improve gastrointestinal hormone disruption, and protect the colon mucus layer. Analysis of 16S rRNA gene sequencing indicated that rhubarb could improve the disorder of intestinal microbiota in constipated rats. For example, beneficial bacteria such as Ligilactobacillus, Limosilalactobacillus, and Prevotellaceae UCG-001 were remarkably increased, and pathogens such as Escherichia-Shigella were significantly decreased after rhubarb treatment. Additionally, the fecal metabolic profiles of constipated rats were improved by rhubarb. After rhubarb treatment, metabolites such as chenodeoxycholic acid, cholic acid, prostaglandin F2α, and α-linolenic acid were markedly increased in constipation rats; in contrast, the metabolites such as lithocholic acid, calcidiol, and 10-hydroxystearic acid were notably reduced in constipation rats. Moreover, correlation analysis indicated a close relationship between intestinal flora, fecal metabolites, and biochemical indices associated with constipation. In conclusion, the amelioration of rhubarb in constipation might modulate the intestinal microflora and its metabolism. Moreover, the application of fecal metabolomics could provide a new strategy to uncover the mechanism of herbal medicines.Key points• Rhubarb could significantly improve gut microbiota disorder in constipation rats.• Rhubarb could markedly modulate the fecal metabolite profile of constipated rats.

Physiological responses of Holstein calves to heat stress and dietary supplementation with a postbiotic from Aspergillus oryzae

Increased ambient temperature causes heat stress in mammals, which affects physiological and molecular functions. We have recently reported that the dietary administration of a postbiotic from Aspergillus oryzae (AO) improves tolerance to heat stress in fruit flies and cattle. Furthermore, heat-induced gut dysfunction and systemic inflammation have been ameliorated in part by nutritional interventions. The objective of this study was to characterize the phenotypic response of growing calves to heat stress compared to thermoneutral ad libitum fed and thermoneutral feed-restricted counterparts and examining the physiologic alterations associated with the administration of the AO postbiotic to heat-stressed calves with emphasis on intestinal permeability. In this report, we expand previous work by first demonstrating that heat stress reduced partial energetic efficiency of growth in control (45%) but not in AO-fed calves (62%) compared to thermoneutral animals (66%). While heat stress increased 20% the permeability of the intestine, AO postbiotic and thermoneutral treatments did not affect this variable. In addition, AO postbiotic reduced fecal water content relative to thermoneutral and heat stress treatments. Heat stress increased plasma concentrations of serum amyloid A, haptoglobin and lipocalin-2, and administration of AO postbiotic did not ameliorate this effect. In summary, our findings indicated that heat stress led to reduced nutrient-use efficiency and increased systemic inflammation. Results suggest that the AO postbiotic improved energy-use efficiency, water absorption, and the intestinal permeability in heat stress-mediated increase in gut permeability but did not reduce heat stress-mediated rise in markers of systemic inflammation.

Sargassum plagiophyllum Extract Enhances Colonic Functions and Modulates Gut Microbiota in Constipated Mice.

Constipation is a symptom that is widely found in the world’s population. Various dietary supplementations are used to relieve and prevent constipation. Seaweed is widely used for its health benefits. In this study, we aimed to investigate the effects of Sargassum plagiophyllum extract (SPE) on functions of the gastrointestinal tract and gut microbiota. The results show that SPE pretreatment increased the frequency of gut contraction, leading to reduce gut transit time. SPE pretreatment also significantly increased the secretion of Cl− and reduced Na+ absorption, increasing fecal water content in constipated mice (p < 0.05). In addition, the Bifidobacteria population in cecal contents was significantly higher in constipated mice pretreated with 500 mg/kg SPE for 14 days than in untreated constipated mice (p < 0.05). Our findings suggest that SPE can prevent constipation in loperamide-induced mice. This study may be useful for the development of human food supplements from S. plagiophyllum, which prevent constipation.

NOVEL INSIGHTS INTO THE MECHANISM OF ACTION OF EXCLUSIVE ENTERAL NUTRITION IN CROHN’S DISEASE: PRE-CLINICAL STUDY IN A RODENT MODEL OF CROHN’S-LIKE COLITIS

Abstract BACKGROUND AND AIMS Exclusive enteral nutrition (EEN) is the only established dietary therapy for Crohn’s disease (CD). However, mechanisms of action of EEN in CD are not clear. CD is characterized by recurrent transmural inflammation in the gut. Mechanical stress, associated with inflammatory cell infiltration, edema, fibrosis, and obstruction, is inevitably present in CD tissue. We hypothesized that transmural inflammation causes mechanical stress, which induces expression of mechanosensitive genes such as IL-6 and COX-2 in the gut, and that EEN treatment attenuates inflammation by alleviating the effect of mechanical stress-induced expression of proinflammatory genes in CD. METHODS CD-like colitis was induced by intracolonic instillation of TNBS (65 mg/kg in 250 μL of 40% ethanol) into the distal colon of 8∼10 weeks old SD rats. Control rats were treated with intracolonic saline instillation. We fed the control and colitis rats with either normal pellet food or EEN with Ensure liquid diet ad lib. Rats were euthanized 7 days after treatments. RESULTS (1) TNBS colitis rats presented decreased body weight, increased disease activity and inflammation index of the colon in pellet food treated rats, which were dramatically improved by EEN treatment. (2) EEN did not significantly affect body weight in control rats, though it dramatically reduced fecal weight by 84.5(±4.3)%, and increased fecal water content by 29.8(±1.3)%. (3) In the solid food fed rats, TNBS treatment induced a localized transmural inflammation with narrowed lumen in the site of instillation (site I) (Figure 1). This led to a distended colon segment (site P) prior to site I, and a non-distended segment (site D) distal to site I. Expression of IL-6 and COX-2 mRNAs was increased significantly (p&amp;lt;0.01) by 111(±44) and 13.1(±2.5)-fold in site I of CD rats, compared to controls. Importantly, the IL-6 and COX-2 mRNA expression was also significantly increased in the distended site P [228(±72) and 16.6(±4.3)-fold], but not in site D (Figure 2). The increased expression of IL-6 and COX-2 occurred mainly in the colonic smooth muscle cells (SMC). (4) However, EEN treatment with Ensure Regular prevented lumen distention in colitis rats. The increase of IL-6 and COX-2 expression in colitis was attenuated drastically in sites I and P (Figure 2). Treatment with other types of liquid diet (Ensure Clear or rodent liquid diet) had similar effects. (5) Expression of IL-6 and COX-2 was highly inducible by mechanical stretch in cultured colonic SMC in vitro and by lumen distention in colon obstruction. CONCLUSIONS Mechanical stress plays a critical role in gut inflammation by inducing expression of mechanosensitive proinflammatory mediators in CD-like colitis. EEN treatment reduces inflammation in CD by alleviating the effects of mechanical stress on expression of genes such as IL-6 and COX-2.

Polydatin improves intestinal barrier injury after traumatic brain injury in rats by reducing oxidative stress and inflammatory response via activating SIRT1-mediated deacetylation of SOD2 and HMGB1

To investigate the protective effect against intestinal mucosal injury in rats following traumatic brain injury (TBI) and explore the underlying mechanism. SD rat models of TBI were established by fluid percussion injury (FPI), and the specimens were collected at 12, 24, 48, and 72 h after TBI. Another 15 rats were randomly divided into shamoperated group (n=5), TBI with saline treatment (TBI+NS) group (n=5), and TBI with PD treatment (TBI+PD) group (treated with 30 mg/kg PD after TBI; n=5). Body weight gain and fecal water content of the rats were recorded, and after the treatments, the histopathology of the jejunum was observed, and the levels of D-lactic acid (D-LAC), diamine oxidase (DAO), ZO-1, claudin-5, and reactive oxygen species (ROS) were detected. Lipid peroxide (LPO) and superoxide dismutase (SOD) 2 content, jejunal pro-inflammatory factors (IL-6, IL-1β, and TNF- α), Sirt1 activity, SOD2 and HMGB1 acetylation level were also determined after the treatments. The rats showed significantly decreased body weight and fecal water content and progressively increased serum levels of D-LAC and DAO after TBI (P < 0.05) with obvious jejunal injury, significantly decreased expression levels of ZO-1 and claudin-5, lowered SOD2 and Sirt1 activity (P < 0.05), increased expression levels of LPO, ROS, and pro-inflammatory cytokines, and enhanced SOD2 and HMGB1 acetylation levels (P < 0.05). Compared with TBI+NS group, the rats in TBI+PD group showed obvious body weight regain, increased fecal water content, reduced jejunal pathologies, decreased D-LAC and DAO levels (P < 0.05), increased ZO-1, claudin-5, SOD2 expression levels and Sirt1 activity, and significantly decreased ROS, LPO, pro-inflammatory cytokines, and acetylation levels of SOD2 and HMGB1 (P < 0.05). PD alleviates oxidative stress and inflammatory response by activating Sirt1-mediated deacetylation of SOD2 and HMGB1 to improve intestinal mucosal injury in TBI rats.

Transdermal Administration of Volatile Oil from Citrus aurantium-Rhizoma Atractylodis Macrocephalae Alleviates Constipation in Rats by Altering Host Metabolome and Intestinal Microbiota Composition.

Background The Citrus aurantium- (ZhiShi, ZS-) Rhizoma Atractylodis Macrocephalae (BaiZhu, BZ) pairs are often found in herbal formulas for constipation. The volatile oils of ZS and BZ (ZBVO) have good pharmacological activity against constipation, but the mechanism for treatment of slow transit constipation (STC) remains unclear. Method A rat model using diphenoxylate tablets was constructed to investigate if transdermal administration of ZBVO would mediate intestinal microorganisms and fecal metabolites and improve STC symptoms. The regulatory effects of ZBVO at 0.15, 0.30, and 0.60 mL kg−1 d−1 on STC rats were assessed by measuring fecal water content, intestinal propulsion rate, histopathology, expression of gastrointestinal hormones, brain and intestinal peptides, and inflammatory factors. The changes in intestinal flora of STC rats were analyzed by 16S rRNA gene sequencing. Moreover, the untargeted fecal metabolomics analysis was performed by ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometer (UPLC-Q-TOF-MS) technology. Results The results showed that ZBVO had a modulating effect on STC by increasing the fecal water content and intestinal propulsion rate. Transdermal administration of ZBVO decreased serum levels of interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) and increased the levels of gastrin (GAS) and substance P (SP). In addition, ZBVO increased 5-hydroxytryptamine (5-HT) levels and decreased vasoactive intestinal peptide (VIP) levels in colon and hippocampus tissues. The results of intestinal microbiota showed that ZBVO improved the diversity and abundance of intestinal microbiota and changed the community composition by decreasing Romboutsia and increasing Proteobacteria, Allobaculum, and Ruminococcaceae. And the feces metabolomics found that nicotinate and nicotinamide metabolism, purine metabolism, citrate cycle (TCA cycle), pyruvate metabolism, arachidonic acid metabolism, pyrimidine metabolism, and primary bile acid biosynthesis were modulated. Conclusion These findings suggest that ZBVO can alleviate STC symptoms by promoting intestinal peristalsis, increasing fecal water content, regulating gastrointestinal hormone level, reducing the inflammatory response, and regulating brain and intestinal peptides after transdermal administration. And structural changes in the intestinal microbiota are closely related to host metabolism and intestinal microbiota destroyed in STC modeling could be significantly improved by the ZBVO, which provides a reference for the development of aromatic drug macrohealth products.

Effect of kefir consumption on intestinal microbiota and some blood parameters inAngora cats

Probiotics are one of the best alternatives to improve gut health. Kefir, which was discovered in the North Caucasus centuries ago, is still consumed frequently today due to its health benefits. Its impacts on the gastrointestinal system have begun to be investigated in animals. In this study, we focused to examine the effect of kefir on intestinal microbiota, some hematological parameters, and fecal quality in Angora cats to provide preliminary data regarding the hypothesis of its use as an alternative probiotic food supplement. Commercial kefir was given orally (30 mL/kg) to seven healthy Angora cats for 14 days. On day 0 and day 14, fresh feces and blood of the cats were collected. The results showed that two-week kefir consumption significantly increased the number of total mesophilic aerobic bacteria, lactococci, lactobacilli, and yeast in the gut microbiota (p < 0.05). Also, a significant decrease was recorded in the number of enterococci (p < 0.05). Measured hematological parameters (WBC, RBC, HGB, PCV, MCV, MCH, MCHC, PLT) were not affected during the experiment (p > 0.05). Among the biochemical parameters (ALT, AST, TP, TG, TC, HDL, LDL, LDH, K, Ca, Na) only a decrement in the activity of LDH, and an increment in K were observed after two-week of kefir consumption (p < 0.05). Additionally, no significant changes were recorded in the body weights, body condition scores, fecal scores, and fecal water contents (p > 0.05). Daily kefir consumption positively altered the intestinal microbiota of Angora cats by increasing the total mesophilic aerobic bacteria, lactococci, lactobacilli, and yeast. Moreover, no detrimental effect was observed in the blood parameters, body condition scores, and fecal quality. Therefore, it could be suggested that including kefir in Angora cats? daily diets can improve their health conditions.

Longitudinal metabolomics profiling of serum amino acids in rotenone-induced Parkinson's mouse model.

Recently, the detailed etiology and pathogenesis of Parkinson's disease (PD) have not been fully clarified yet. Increasing evidences suggested that the disturbance of peripheral branched-chain amino acids (BCAAs) metabolism can promote the occurrence and progression of neurodegenerative diseases through neuroinflammatory signaling. Although there are several studies on the metabolomics of PD, longitudinal study of metabolic pathways is still lacking. Therefore, the purpose of the present study was to determine the longitudinal alterations in serum amino acid profiles in PD mouse model. Gas chromatography-mass spectrometry (GC-MS) was applied to detect serum amino acid concentrations in C57BL/6 mice after 0, 3 and 4weeks of oral administration with rotenone. Then the data were analysed by principal component analysis (PCA) and orthogonal projection to latent structures (OPLS) analysis. Finally, the correlations between different kinds of serum amino acids and behaviors in rotenone-treated mice were also explored. Compared with 0-week mice, the levels of L-isoleucine and L-leucine were down-regulated in 3-week and 4-week mice, especially in 4-week mice. Moreover, the comprehensive analysis showed that L-isoleucine and L-leucine were negatively correlated with pole-climbing time and positively correlated with fecal weight and water content of PD mice. These results not only suggested that L-isoleucine and L-leucine may be potential biomarkers, but also pointed out the possibility of treating PD by intervening in the circulating amino acids metabolism.