Changes in the Gut Microbiome after Galacto-Oligosaccharide Administration in Loperamide-Induced Constipation

Unbalanced dietary habits and the consumption of high protein and instant foods cause an increase in constipation. Here, we evaluated the effects of galacto-oligosaccharide (GOS) on a rat model of loperamide-induced constipation by measuring various biological markers and cecal microbiota. The fecal water content and intestinal transit ratio significantly increased in the GOS-administered (GL and GH) groups than in the control group (p < 0.05, p < 0.01, and p < 0.001, respectively). The length of intestinal mucosa (p < 0.05 and p < 0.01, respectively) and area of crypt cells were (p < 0.01, both) significantly increased in the GOS-administered groups compared to the control group. The distribution of interstitial cells of Cajal, which is related to the intestinal movement, showed a significant increase in GOS-administered groups than in the control group (p < 0.01, both). The relative abundance of lactic acid bacteria (LAB), especially Lactobacillus and Lactococcus, significantly increased in the GL group than in the control group. Furthermore, there was a positive correlation between short chain fatty acids (SCFAs) and the gut microbiota in the GL groups. These results demonstrated that GOS administration effectively alleviates constipation by increasing LAB proliferation in the intestinal microbiota and SCFA production.

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Effect of Chaihu Shugan Powder-Contained Serum on Glutamate-Induced Autophagy of Interstitial Cells of Cajal in the Rat Gastric Antrum

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/7318616 ◽

2019 ◽

Vol 2019 ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Ren-Qian Tan ◽

Zhi Zhang ◽

Jing Ju ◽

Jiang-Hong Ling

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Interstitial Cells Of Cajal ◽

Underlying Mechanism ◽

Control Group ◽

Motility Disorders ◽

Transmission Electron ◽

Cells Of Cajal ◽

Bcl2 Expression ◽

Gi Motility

Gastrointestinal (GI) motility disorder is caused by excessive autophagy of the interstitial cells of Cajal (ICC). Chaihu Shugan Powder (CSP) is a traditional Chinese medicine with therapeutic benefits in GI motility disorders; however, the underlying mechanism of its therapeutic effect in GI disorders, especially autophagy of ICC, remains unclear. Thus, this study investigated the effects of CSP-contained serum on glutamate-induced autophagy in rat gastric ICC, exploring its underlying mechanism. In vitro cultured rat stomach ICC were identified by fluorescence microscopy and then stimulated with glutamate (5 mmol/L) for 3 h to establish the autophagy model. These cells were then treated with 10% CSP-containing serum or the autophagy inhibitor 3-methyladenine (3-MA; 5 mmol/L) for 24 h. The control group was cultured with only 10% serum containing physiological saline. The viability of ICC was measured by the CCK-8 assay. The ultrastructure and autophagosomes of ICC were observed using transmission electron microscopy. LC3 expression was detected by immunofluorescence, and LC3, Beclin1, Bcl2, and PI3KC3 expression was detected by western blot analysis. Transmission electron microscopy showed abundant endoplasmic reticulum, mitochondria, and other organelles in the control group, whereas the cells in the autophagy model control group had clear autophagic vacuoles, which were not apparent in both CSP and 3-MA groups. ICC viability was significantly increased by CSP and 3-MA interventions (P < 0.01), accompanied by a decrease in LC3 fluorescence (P < 0.01). Moreover, the expression levels of LC3II/I, Beclin1, and PI3KC3 were significantly decreased (all P < 0.01) with CSP and 3-MA treatment, while Bcl2 expression level was higher than that of the model group (P < 0.01). Thus, CSP can reduce autophagic damage by enhancing Bcl2 expression and downregulating the expression of LC3, Beclin1, and PI3KC3 to protect ICC. These results highlight the potential of CSP in the treatment of GI motility disorders.

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Acute Cholecystitis Reduces Interstitial Cells of Cajal in Porcine Gallbladder Through Decreased mRNA Synthesis

Cellular Physiology and Biochemistry ◽

10.1159/000489987 ◽

2018 ◽

Vol 47 (2) ◽

pp. 535-544 ◽

Cited By ~ 4

Author(s):

Zhen-peng Huang ◽

Hu Qiu ◽

Bao-ping Yu

Keyword(s):

Acute Cholecystitis ◽

Guinea Pigs ◽

Interstitial Cells Of Cajal ◽

Interstitial Cells ◽

Motility Disorder ◽

Gallbladder Motility ◽

Control Group ◽

Mrna Levels ◽

Duct Ligation ◽

Cells Of Cajal

Background/Aims: Acute cholecystitis is a common gastrointestinal disorder, often characterized by acute cholecystitis with gallbladder motility disorder. Interstitial cells of Cajal (ICCs) are the pacemaker cells of gut motility in the gastrointestinal tract. Disruption of ICC function is related to motility disorders. The aim of this study was to explore the cellular and molecular mechanisms of ICCs in acute cholecystitis and after the resolution of acute inflammation. Materials and Methods: Fifty adult guinea pigs were randomly divided into five groups: a sham-administered group (control group); two groups that were intraperitoneally administered an anti-polyclonal neutrophil (PMN) antibody 24 h before common bile duct ligation (CBDL); and two groups of guinea pigs that were subjected to CBDL without receiving the PMN antibody. Guinea pigs that underwent CBDL were held for 24 h or 48 h after surgery before being subjected to laparotomy and cholecystectomy. Immunohistochemistry, TUNEL assays, western blotting, and real-time PCR were performed to determine ICC morphology and density, to detect ICC apoptosis, and to examine stem cell factor (SCF) and c-kit protein expression and SCF and c-kit mRNA levels, respectively. Results: Both hematoxylin-eosin staining and histological inflammation scores in the PMN groups were lower than those in the control groups (P < 0.01). No differences were observed in ICC morphology between groups. During acute cholecystitis, ICCs numbers were reduced. Conversely, the density of ICCs increased after inflammation was relieved (P < 0.01). In addition, SCF and c-kit protein and mRNA expression levels decreased during acute cholecystitis (P < 0.05) and increased after inflammation was relieved (P < 0.05). Furthermore, ICC apoptosis increased during acute cholecystitis and decreased after resolution of acute cholecystitis (P < 0.01). Conclusions: In acute cholecystitis, ICC injury may be related to gallbladder motility disorder.

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Electroacupuncture at ST36 Protects ICC Networks via mSCF/Kit-ETV1 Signaling in the Stomach of Diabetic Mice

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2017/3980870 ◽

2017 ◽

Vol 2017 ◽

pp. 1-13 ◽

Cited By ~ 3

Author(s):

Lugao Tian ◽

Beibei Zhu ◽

Shi Liu

Keyword(s):

Gastric Emptying ◽

High Frequency ◽

Interstitial Cells Of Cajal ◽

Low Frequency ◽

Control Group ◽

Diabetic Patients ◽

Diabetic Mice ◽

Protein Levels ◽

Reliable Marker ◽

Cells Of Cajal

Background. Electroacupuncture (EA) at ST36 has been used to regulate gastric motility and effectively improve gastric emptying in diabetic patients. Nevertheless, the specific mechanisms underlying the efficacy of this treatment remain unknown. The aim of this study was to assess the variations of interstitial cells of Cajal (ICC) and explore the changes in mSCF/KIT-ETV1 signaling in the antrum and corpus of diabetic mice after treatment with EA.Methods. Male C57BL/6 mice were randomized into five groups: control group, diabetic group (DM), diabetic-plus-sham EA group (SEA), diabetic-plus-low-frequency EA group (LEA), and diabetic-plus-high-frequency EA group (HEA). The expression levels of Ano1, c-Kit, and ETV1 were assessed by immunofluorescence in the antrum and corpus. Western blotting and PCR methods were further used to evaluate c-Kit, mSCF, and ETV1 expression.Results. (1) c-Kit and Ano1 were obviously decreased in the DM group, but c-Kit reduced much more than Ano1. (2) The mSCF, c-Kit, and ETV1 mRNA and protein levels were obviously decreased in the DM group in both the antrum and the corpus (P<0.01), but they were significantly elevated in the LEA and HEA groups (P<0.01).Conclusions. Ano1 is a reliable marker to detect ICC changes in diabetes; low- and high-frequency EA at acupoint ST36 can protect the networks of ICC possibly via normal activation of mSCF/KIT-ETV1 signaling.

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Long-Pulse Gastric Electrical Stimulation Repairs Interstitial Cells of Cajal and Smooth Muscle Cells in the Gastric Antrum of Diabetic Rats

Gastroenterology Research and Practice ◽

10.1155/2018/6309157 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 4

Author(s):

Yan Chen ◽

Hongcai Wang ◽

Hai Li ◽

Shi Liu

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Interstitial Cells Of Cajal ◽

Muscle Cells ◽

Interstitial Cells ◽

Diabetic Rats ◽

Control Group ◽

Cells Of Cajal ◽

Gastric Contractions ◽

Long Pulse

Background/Aims. The damage of interstitial cells of Cajal and smooth muscle cells has far-reaching implications in the pathogenesis of gastroparesis in diabetic patients. Gastric electrical stimulation (GES) is an efficient therapy for gastric motility disorders, but the mechanisms of GES require clarification.Methods. Male rats were randomly divided into the control group, diabetic rat group (DM), diabetic rats with sham GES group (DM + SGES), and diabetic rats with different frequency GES group (DM + GES) (GES1: 5.5 cpm, 100 ms, 4 mA; GES2: 5.5 cpm, 300 ms, 4 mA; and GES3: 5.5 cpm, 550 ms, 2 mA). Gastric contractions were explored using the organ bath technique. The alterations of interstitial cells of Cajal, the SCF/c-kit pathway, and smooth muscle cells were also investigated.Results. (1) Gastric contractions were significantly improved in the DM + GES group compared with those in the DM group. (2) The damage of interstitial cells of Cajal was prevented in the DM + GES group in contrast to the DM group. Moreover, long-pulse GES increased the expression of the SCF/c-kit pathway. More proliferated interstitial cells of Cajal in muscle layers were observed obviously in the DM + GES group. (3) The number of smooth muscle cells in the DM group was not significantly decreased compared with that in the control group. However, ultrastructural changes were distinctly damaged in the DM group. The application of GES protected against the alteration of the ultrastructures of smooth muscle cells.Conclusions. Long-pulse GES improves gastric contraction possibly by enhancing the proliferation of interstitial cells of Cajal and restoring the injury of smooth muscle cells.

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Electroacupuncture Relieves Suppression of Autophagy in Interstitial Cells of Cajal of Diabetic Gastroparesis Rats

Canadian Journal of Gastroenterology and Hepatology ◽

10.1155/2020/7920715 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Xing Wei ◽

Yaping Lin ◽

Dongfeng Zhao ◽

Xiaojuan Xiao ◽

Qiao Chen ◽

...

Keyword(s):

Blood Glucose ◽

Gastric Emptying ◽

Interstitial Cells Of Cajal ◽

Interstitial Cells ◽

Diabetic Gastroparesis ◽

Control Group ◽

Autophagy Flux ◽

Gastric Emptying Rate ◽

Modeling Group ◽

Cells Of Cajal

Background. The incidence of diabetic gastroparesis (DGP) is mainly blamed to abnormity of interstitial cells of Cajal (ICCs). Autophagy could degrade damaged proteins and organelles to keep intracellular homeostasis, and it could directly influence structure and number of cells. In this study, we aimed to figure out the relationship between DGP and autophagy of ICCs. Methods. Sixty Sprague-Dawley (SD) rats were randomly divided into normal control group (NC, 10) and modeling group (50). Rats in the modeling group were injected 2% streptozotocin (STZ) and fed with high-glucose and high-fat diet for 8 weeks in order to establish DGP rat model. After modeling, 30 successfully modeled rats were randomly selected and separated into diabetic gastroparesis group (DGP, 10), GDP rats with electroacupuncture group (EA, 10), and GDP rats with metoclopramide group (MP, 10). When the intervention was completed, blood glucose was measured by ONE TOUCH glucometer and gastrointestinal propulsive rate was detected through measuring optical density. Autophagosomes were observed under transmission electron microscope (TEM). The expression of LC3 protein and P62 protein was measured by Western blot. When ICCs were transfected with GFP-RFP-LC3 plasmid, autophagy flux was observed by laser scanning confocal microscope. Results. (1) After intervention, compared with blood glucose of rats in the NC group, all of the DGP, EA, and MP groups were remarkably increased (P<0.01); compared with the DGP group, the blood glucose of the EA and MP groups was decreased greatly (P<0.01). (2) Compared with gastrointestinal propulsive rate of rats in the NC group, no matter gastric emptying rate or intestinal propulsive rate, the EA and MP groups were significantly reduced (P<0.01); compared with the NC group, gastric emptying rate and intestinal propulsive rate in the EA group were obviously decreased (P<0.05, P<0.01); compared with the DGP group, the EA and MP groups were increased significantly (P<0.01). (3) Compared with the NC group, intensity of RFP and GFP in the DGP group was obviously increased (P<0.05, P<0.01), in other words, the DGP group accompanying suppression of autophagy; compared with the DGP group, intensity of RFP and GFP in the EA group was decreased significantly (P<0.05, P<0.01). (4) There was no autophagosome in the NC group, and an autophagosome existed in the DGP group. Both EA and MP groups found autophagy. (5) When coming to LC3 II/LC3 I, compared with the NC group, the ratio was enhanced in the DGP and EA groups (P<0.01, P<0.05); compared with the DGP group, LC3 II/LC3 I was dramatically decreased in the MP and EA groups (P<0.01). (6) As the substrate of degradation, the expression of P62 in the other three groups was significantly increased (P<0.01) compared with the NC group; compared with the DGP group, the amount of P62 in the EA and MP groups was reduced greatly (P<0.01). Conclusion. The impaired autophagy flux in ICCs is the pathological basis of diabetic gastroparesis, blaming to fusion dysfunction of autophagosome and lysosome and electroacupuncture (EA) could ease the suppression of autophagy to improve gastric motility.

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Positive Influence of a Probiotic Mixture on the Intestinal Morphology and Microbiota of Farmed Guinea Fowls (Numida meleagris)

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.743899 ◽

2021 ◽

Vol 8 ◽

Author(s):

Livio Galosi ◽

Salvatore Desantis ◽

Alessandra Roncarati ◽

Patrizia Robino ◽

Alessandro Bellato ◽

...

Keyword(s):

Short Chain Fatty Acids ◽

Treated Group ◽

Positive Influence ◽

Intestinal Morphology ◽

Control Group ◽

Numida Meleagris ◽

Cecal Microbiota ◽

Anti Inflammatory ◽

Stability And Resilience ◽

Microbiota Diversity

To understand the effectiveness of a probiotic mixture on intestinal morphology, mucus layer composition, and cecal microbiota diversity, 40 10-day-old Guinea fowls (Numida meleagris) were assigned to two groups: the control group (C), receiving drinking water, and the treated group (P), receiving water plus a commercial multi-strain probiotic (Slab51®, 2 × 1011 CFU/L). Birds were slaughtered after 4 months, and the intestines were collected. Samples from the duodenum, ileum, and cecum were processed for morphological and morphometric studies, and conventional glycohistochemistry. Cecal samples were also used to assess the microbiota by 16S metataxonomic approach. Group P showed significant increase in the villus height (p < 0.001 in the duodenum and p < 0.05 in the ileum and cecum), villus width (p < 0.05 in all investigated tracts), depth of crypts (p < 0.001 in the duodenum and cecum; p < 0.05 in the ileum), and goblet cells per villus (p < 0.001 in all investigated tracts) compared with group C. Cecal microbiota of the birds varied considerably and comparing the relative abundance of the main observational taxonomic units (OTUs), a positive enrichment of several beneficial taxa, such as Oscillospira, Eubacterium, Prevotella, and members of the Ruminococcaceae, was observed. The enrichment of those taxa can improve microbiota stability and resilience facing environmental stresses, enhancing its resistance against invading pathogens. Ruminococcaceae, which represent the most important taxon in both groups, and Prevotella have a key role in the gut physiology due to the production of short-chain fatty acids (SCFAs), which are a vital energy source for enterocytes, improve glucose metabolism, and exert an overall anti-inflammatory effect. Probiotic administration enriches the presence of Coprococcus, Oscillospira, and Eubacterium taxa that produce butyrate, which exerts a beneficial effect on growth performance, structure of villi, and pathogen control and has anti-inflammatory properties too. This study indicates that Slab51® supplementation positively affects the morphology and microbiota diversity of the guinea fowl intestine.

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