Small Intestinal Mucosal Cells Research Articles

Valproic acid alleviates total-body irradiation-induced small intestinal mucositis in mice

Purpose Gastrointestinal (GI) injury is one of the serious problems of total-body irradiation (TBI). However, no fundamental treatment for TBI and other radiation-induced GI injury has yet been established. Valproic acid (VPA) administration reduces mortality in mice subjected to total-body irradiation (TBI) with X-rays. This study aimed to evaluate the effects of VPA on GI injury induced by TBI in mice. Materials and methods Mice were subjected to TBI with X-rays to induce GI injury. Changes in survival and weight were observed after VPA administration. The small intestine was then sampled at 0, 1, 3, 7, and 10 d after irradiation for histological and immunohistological evaluation and measurement of myeloperoxidase (MPO) activity and inflammatory cytokine levels (IL-1β). Results VPA (200 and 600 mg/kg) increased survival rate and reduced weight loss in model mice. IL-1β expression 1 d after irradiation was significantly lower in the VPA group than that in the vehicle group. Furthermore, the increase in MPO activity at 3 and 7 d after irradiation was significantly suppressed by VPA administration. Histological examination (hematoxylin and eosin staining) revealed that 600 mg/kg VPA inhibited inflammatory cell infiltration. Immunostaining for the proliferating cell nuclear antigen involved in cell proliferation showed that VPA suppressed the irradiation-induced decrease in cell proliferative capacity. Conclusions Treatment with VPA in mice with GI injury caused by TBI suppressed inflammatory responses in small intestinal mucosal cells. These results suggest that VPA may be a useful therapeutic agent against TBI-induced small intestinal mucositis.

Nutritional Approach of Neonatal with High Output Stoma Due to Long Segment Hirschsprung Disease: A Case Report

Background: High Output Stoma (HOS) continues to be one of the most challenging problems for pediatrician especially in neonates. One of the most common causes in neonatal HOS is post resection long segment Hirschsprung disease.
 Case: We reported a case of three-day-old baby boy diagnosed as Hirschsprung diseases with peritonitis possibility and did laparotomy with ileal resection, double barrel ileostomy and biopsy. Nutritional management is a major subject on taking care of this type of neonatal patient. We share our experience in limited facilities with all the patient uniqueness
 Discussion: Loss of a significant length of the small bowel results in interrelated physiologic events as a result of decreased small intestinal mucosal absorptive cell. This leads to a lesser fraction of ingested food and intestinal secretion thus causing an excessive volume loss. The introduction of early enteral feeds promotes intestinal adaptation, with subsequent weaning off parenteral nutrition. Most off patient with high output stoma will require parenteral nutrition which is associated with acute and long-term complications. In our case, we did early nutritional intervention using parenteral and enteral nutrition, counting ongoing fluid loss trough stoma and adjust it to total daily fluid requirement. We found weight loss during hospitalized due to HOS, and we do catch up at the end. We found difficulties to adjust comparation between enteral and parenteral intake to maintain the weight gain.
 Conclusion: Although parenteral nutrition is often necessary, at least initially, the therapeutic goal should be to enhance intestinal adaptation and enteral nutrient assimilation, and thereby reduce parenteral nutrition requirements. Daily monitoring for ongoing fluid loss very crucial for adjusting nutrition.

Blocking TRAIL-DR5 signaling pathway with soluble death receptor 5 fusion protein mitigates radiation-induced injury.

The increasing application of nuclear technology, the high fatality of acute radiation syndrome (ARS) and its complex mechanism make ARS a global difficulty that requires urgent attention. Here we reported that the death receptor 5 (DR5), as well as its ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), were both significantly upregulated after irradiation in mice with 6 Gy γ-ray single radiation. And by intravenously administrated with soluble DR5 fusion protein (sDR5-Fc), the competitive antagonist of DR5, the excessive apoptosis in the radiation-sensitive tissues such as spleen and thymus were significantly inhibited and the radiation-induced damage of spleen and thymus were mitigated, while the expression of apoptosis-inhibiting proteins such as Bcl-2 was also significantly upregulated. The biochemical indicators such as serum ALP, AST, ALT, TBIL, K, and Cl levels that affected by radiation, were improved by sDR5-Fc administration. sDR5-Fc can also regulate the number of immune cells and reduce blood cell death. For in vitro studies, it had been found that sDR5-Fc effectively inhibited apoptosis of human small intestinal mucosal epithelial cells and IEC-6 cells using flow cytometry. Finally, survival studies showed that mice administrated with sDR5-Fc after 9 Gy γ-ray single whole body radiation effectively increased the 30-day survival and was in a significant dose-dependent manner. Overall, the findings revealed that DR5/TRAIL-mediated apoptosis pathway had played important roles in the injury of ARS mice, and DR5 probably be a potential target for ARS therapeutics. And the DR5 apoptosis antagonist, sDR5 fusion protein, probably is a promising anti-ARS drug candidate which deserves further investigation.

Beware of accidental ingestion of Colchicum autumnale mistaken for Allium victorialis.

Beware of accidental ingestion of Colchicum autumnale mistaken for Allium victorialis.

Dietary supplementation ellagic acid on the growth, intestinal immune response, microbiota, and inflammation in weaned piglets.

Piglets are susceptible to weaning stress, which weakens the barrier and immune function of the intestinal mucosa, causes inflammation, and ultimately affects animal growth and development. Ellagic acid (EA) is a natural polyphenol dilactone with various biological functions. However, The mechanisms underlying the effects of EA on animal health are still poorly known. Herein, we examined whether dietary supplementation with EA has a positive effect on growth performance, intestinal health, immune response, microbiota, or inflammation in weaned piglets. Sixty weaned piglets (age, 30 days) were randomly divided into two groups: the control group (basic diet) and the test group (basic diet + 500 g/t EA). The pigs were fed for 40 days under the same feeding and management conditions, and the growth performance of each individual was measured. At the end of the feeding period, samples were collected from the small intestinal mucosa for further analysis. Using these tissues, the transcriptome sequences and intestinal microbial diversity were analyzed in both groups. An inflammation model using small intestinal mucosal epithelial cells (IPEC-J2) was also constructed. Dietary EA supplementation significantly increased the average daily weight gain (ADG) and reduced diarrhea rate and serum diamine oxidase (DAO) levels of weaned piglets. Transcriptome sequencing results revealed 401 differentially expressed genes in the jejunum mucosal tissue of pigs in the control and test groups. Of these, 163 genes were up-regulated and 238 were down-regulated. The down-regulated genes were significantly enriched in 10 pathways (false discovery rate < 0.05), including seven pathways related to immune response. The results of bacterial 16s rDNA sequencing show that EA affects the composition of the intestinal microbiota in the cecum and rectum, and reveal significant differences in the abundances of Prevotella_9, Lactobacillus delbrueckii, and Lactobacillus reuteri between the test and control groups (P < 0.05). Experiments using the inflammation model showed that certain doses of EA promote the proliferation of IPEC-J2 cells, increase the relative mRNA expression levels of tight junction-related proteins (ZO-1 and Occludin), improve the compactness of the intestine, reduce the expression of inflammatory factors TNF-α and IL-6, and significantly reduce LPS-induced inflammation in IPEC-J2 cells. In conclusion, we found for the first time that dietary supplementation of EA affects the gut immune response and promotes the beneficial gut microbiota in weaned piglets, reduces the occurrence of inflammatory responses, and thereby promotes the growth and intestinal health of piglets.

PKG II secreted via the classical endoplasmic reticulum-Golgi apparatus secretory pathway blocks the activation of EGFR through phosporalting its threonine 406 and has an anti-tumor effect.

Protein kinase G type II (PKG II) is a serine/threonine-protein kinase that was originally isolated from the small intestinal mucosa with primary functions in the secretion of small intestinal mucosal cells, secretion of renin and aldosterone, and chondrocyte activities. Recent studies have shown that PKG II exerts anti-tumor effects, while a previous study by our group confirmed that PKG II inhibited the proliferation and migration of cancer cells. Interestingly, PKG II, which was typically bound to the intracellular side of the membrane, was detected in the serum and cell culture medium as a diagnostic biomarker of tumor growth. Thus, the aim of the present study was to elucidate the function and the targets of PKG II, and the mechanism underlying the secretion of this kinase. Construction of peptides and plasmids, RNA interference, Immunoelectron microscopy, Co-immunoprecipitation, N-glycosylation assay and Isolation of the Golgi apparatus were applied to investigate the secretory mechanism, and the targets and function of PKG II. PKG II was secreted by enterochromaffin (EC) cells, which were components of the endocrine system in the gastrointestinal tract. Myristoylation of glycine 2 and the N-terminal sequence, especially the amino acids 3-30, acted as a signal peptide to induce the secretion of PKG II via the conventional protein secretory pathway. Moreover, recombinant PKG II inhibited the epidermal growth factor (EGF)-induced activation of the EGF receptor via phosphorylating the T406 of the extracellular domain and blocked EGF-triggered proliferation of various cancer cells. These results revealed a correlation between the endocrine system and the secretion of protein kinase, suggesting a novel protein secretory pathway. The resuls also indicated that secreted PKG II was a potential diagnostic biomarker and an inhibitor of tumor.

Small intestinal mucosal cells in piglets fed with probiotic and zinc: a qualitative and quantitative microanatomical study.

Probiotics and zinc are commonly used and beneficial in pig production. This work aimed to assess the effects of probiotic and zinc on the mucosal cells of the small intestine in respect to digestive capacity and immunity in pre- and post-weaned piglets. Eighteen Large White Yorkshire piglets were divided equally into control and treatment groups. The piglets were maintained in standard management conditions and were weaned at 28 days of age. The treatment group of piglets fed a mixture of probiotics orally at 1.25 × 109 CFU/day and zinc at 2000 ppm/day from birth to 10 days of age. At three different age-groups viz. day 20 (pre-weaning) and, day 30 and day 60 (post-weaning), the animals were sacrificed. For histomorphology, the tissue samples were processed and stained with Mayer's haematoxylin and eosin for routine study, combined periodic acid-Schiff-Alcian blue for mucopolysaccharides and Masson-Hamperl argentaffin technique for argentaffin cells. The stained slides were observed under the microscope. The samples were processed as per the standard procedure for scanning and transmission electron microscopy. The statistical analysis of the data using the appropriate statistical tests was also conducted. The mucosal epithelium of villi and crypts were lined by enterocytes, goblet cells, argentaffin cells, microfold (M-cell) cells, tuft cells and intraepithelial lymphocytes. The multipotent stem cells were located at the crypt base. The length of the enterocyte microvilli was significantly longer (p < 0.05) in the treatment group of piglets. The number of different types of goblet cells and argentaffin cells was more in treated piglets irrespective of segments of intestine and age. The intraepithelial lymphocytes were located in apical, nuclear and basal positions in the lining epithelium of both villus tip and base with their significant increase in the treatment group of piglets. The transmission electron microscopy revealed the frequent occurrence of tuft cells in the lining mucosa of the small intestine in treated piglets. Dietary supplementation of probiotic and zinc induced the number of different mucosal cells of villi and crypts in the small intestine that might suggest the greater absorptive capacity of nutrients and effective immunity in critical pre and post-weaned piglets.

Rat Small Intestinal Mucosal Epithelial Cells Absorb Dietary 1,3-Diacylglycerol Via Phosphatidic Acid Pathways.

Compared with triacylglycerol (TAG), dietary 1,3-diacylglycerol (1,3-DAG) is associated with reduced serum lipid and glucose levels. We investigated the metabolism of 1,3-DAG by assaying its intermediate metabolites during digestion and absorption in the rat small intestine. After gavage with TAG emulsion, TAG was digested mainly to 2-monoacylglycerol (2-MAG) and unesterified fatty acid (FFA) in the rat small intestinal lumen. 2-MAG was directly absorbed into the small intestinal epithelial cells and esterified to 1,2(2,3)-DAG, and further esterified to TAG. After gavage with 1,3-DAG emulsion, 1,3-DAG was digested mainly to 1(3)-MAG and FFA in the rat small intestinal lumen with subsequent significant increase of 1-MAG and 1,3-DAG concentrations in small intestinal mucosal epithelial cells, and the 2-MAG, 1,2(2,3)-DAG, and TAG concentrations in mucosal epithelial cells were not significantly different after 1,3-DAG than after TAG gavage, suggesting that the metabolic pathway of 1,3-DAG is different from that of TAG. In intestinal mucosal epithelial cells, we further assayed enzyme levels and gene expression of proteins in the phosphatidic acid (PtdOH) pathway. The glycerol kinase, phosphatidate phosphatase, and diacylglycerol acyltransferase-2 expression and the relative expression of mRNA of enzymes were significantly increased in the 1,3-DAG group compared with the TAG group, suggesting that TAG synthesis from dietary 1,3-DAG was mainly via PtdOH pathways, which may partially account for the effect of dietary DAG on postprandial serum TAG.

Hypoxia preconditioning protects Ca2+-ATPase activation of intestinal mucosal cells against R/I injury in a rat liver transplantation model.

AIMTo investigate the effect of ischaemia and reperfusion (I/R) injury on the Ca2+-ATPase activation in the intestinal tissue of a rat autologous orthotopic liver transplantation model and to determine if hypoxia preconditioning (HP) therapy induces HIF-1α to protect rat intestinal tissue against I/R injury.METHODSRats received non-lethal hypoxic preconditioning therapy to induce HIF-1α expression. We used an autologous orthotopic liver transplantation model to imitate the I/R injury in intestinal tissue. Then, we detected the microstructure changes in small intestinal tissues, Ca2+-ATPase activity, apoptosis, and inflammation within 48 h postoperatively.RESULTSHIF-1α expression was significantly increased in intestinal tissue at 12 h postoperatively in rats that were exposed to a hypoxic environment for 90 min compared with a non-HP group (HP vs AT, P = 0.0177). Pathological analysis was performed on the intestinal mucosa cells, and the cells in the HP group appeared healthier than the cells in the AT group. The Ca2+-ATPase activity in the small intestinal cells in the AT group was significantly lower after the operation, and the Ca2+-ATPase activity in the HP group recovered faster than that in the AT group at 6 h postoperatively (HP vs AT, P = 0.0106). BCL-2 expression in the HP group was significantly higher than that in the AT group at 12 h postoperatively (HP vs AT P = 0.0010). The expression of the inflammatory factors NO, SOD, IL-6, and TNF-α was significantly lower in the HP group than in the AT group.CONCLUSIONHypoxia-induced HIF-1α could protect intestinal mucosal cells against mitochondrial damage after I/R injury. HP could improve hypoxia tolerance in small intestinal mucosal cells and increase Ca2+-ATPase activity to reduce the apoptosis of and pathological damage to intestinal cells. HP could be a useful way to promote the earlier recovery of intestinal function after graft procedure.

Effect of curcumin on intestinal mucosal mechanical barrier in rats with non-alcoholic fatty liver disease

Objective: To investigate the effect of curcumin on intestinal mucosal mechanical barrier in rats with non-alcoholic fatty liver disease. Methods: A total of 30 male Sprague-Dawley rats were equally divided into normal control group, model group, and curcumin intervention group. The rats in the model group and the curcumin intervention group were given high-fat feed for 16 weeks, and those in the curcumin intervention group were given curcumin 200 mg/kg/day by gavage once a day after 8 weeks of high-fat feeding. The rats were sacrificed at the end of week 16. A light microscope was used to observe pathological changes in the liver, an electron microscope was used to observe the tight junction of the intestinal mucosa, an automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), chromogenic substrate Limulus amebocyte lysate assay was used to measure plasma lipopolysaccharide (LPS) level, spectrophotometric method was used to measure the activity of serum diamine oxidase, ELISA was used to measure the serum level of tumor necrosis factor-α (TNFα), and immunohistochemistry was used to measure the expression of the tight junction protein occludin. One-way ANOVA test and SNK-q test were used for statistical analysis. Results: Under the light microscope, the control group had no hepatocyte steatosis, the model group had significant hepatocyte steatosis and inflammatory cell infiltration, and the curcumin intervention group had reduced hepatocyte steatosis and inflammatory cell infiltration. Under the electron microscope, the control group had a clear and complete structure of the tight junction of the intestinal mucosa and normal structures of mitochondria and endoplasmic reticulum; in the model group, the structure of the tight junction of the intestinal mucosa was destroyed, the intercellular space was widened, the desmosomes had a loose structure, there was edema in some mitochondria, and the endoplasmic reticulum was dilated; the curcumin intervention group had improvements in the structure of tight junction of the intestinal mucosa, intercellular space, edema in the mitochondria, and dilation of the endoplasmic reticulum. Compared with the control group, the model group had significant increases in the serum levels of AST, ALT, DAO, TNFα, and LPS (q = -15.918, -14.402, -33.700, -8.944, and -10.832, P < 0.05); compared with the model group, the curcumin intervention group had significant reductions in the serum levels of AST, ALT, DAO, TNFα, and LPS (q = 10.457, 7.752, 18.802, 5.202, and 4.279, P < 0.05). In the control group, occludin showed a linear distribution along the top of small intestinal mucosal epithelial cells. The model group had a significant reduction in positive staining compared with the control group, and the curcumin intervention group had a significant increase in positive staining compared with the model group. The relative expression of occludin was 0.29±0.03 in the control group, 0.12±0.02 in the model group, and 0.21±0.02 in the curcumin intervention group (P < 0.05). Conclusion: Intestinal mucosal mechanical barrier is impaired in rats with NAFLD. Curcumin can reduce such damage, and its mechanism of action may be related to up-regulating the expression of occludin in the intestinal mucosa and reducing the levels of TNFα and LPS.

Expression of vascular adhesion protein-1 in severe hemorrhagic shock and resuscitation in rats

Objective To observe the expression and activity changes of vascular adhesion protein-1(VAP-1)in the small intestine and serum of rats during severe hemorrhagic shock and resuscitation,and to study its influence on shock prognosis.Methods Fifty rats were evenly randomized into sham group,hemorrhagic shock group,shock resuscitation group,control recovery group and the experimental recovery group.Rat models of severe hemorrhagic shock and resuscitation were established.Before shock,1hour after shock and 1hour after resuscitation,the expressions of VAP-1protein and mRNA in the intestinal tissues of rats were examined by Western blotting analysis and real-time RT-PCR,respectively;and the serum levels of VAP-1and its activities were determined by ELISA kit.Rats in the experimental recovery group was resuscitated by injection of 20mg/kg2-bromoethylamine and those in the control recovery group were given 1 mL/kg normal saline,and then the blood pressure,intestinal mucosa injury(Chiu's score),small intestinal epithelial cell apoptosis(TUNEL detection)and 24-hour survival rates were compared between the two recovery groups.Results The intestinal VAP-1protein and mRNA expressions and the serum VAP-1and its activities in the severe hemorrhagic shock group were significantly higher than those in the sham shock group(P0.05).Compared with the shock group,the above parameters were decreased in the recovery group,but were still higher than those in the sham group.Compared with the saline control group,20mg/kg 2-bromoethylamine significantly increased the blood pressure of animals 1hand 24 hafter recovery(P=0.010,0.039),significantly improved the Chiu's score and apoptosis index of small intestinal epithelial cells(P=0.022,P=0.002),and improved the 24-hour survival rates of rats(90%to 60%).Conclusion The levels of VAP-1and its activities are increased in severe hemorrhagic shock rats,and fluid resuscitation can inhibit this increase.Inhibition of VAP-1activities can improve the low blood pressure,intestinal mucosa injury and apoptosis of small intestinal mucosa cells after the severe hemorrhagic shock and resuscitation,improving the 24-hour survival rates of rats.

Endothelial Binding of Beta Toxin to Small Intestinal Mucosal Endothelial Cells in Early Stages of Experimentally Induced Clostridium Perfringens Type C Enteritis in Pigs

Beta toxin (CPB) is known to be an essential virulence factor in the development of lesions of Clostridium perfringens type C enteritis in different animal species. Its target cells and exact mechanism of toxicity have not yet been clearly defined. Here, we evaluate the suitability of a neonatal piglet jejunal loop model to investigate early lesions of C. perfringens type C enteritis. Immunohistochemically, CPB was detected at microvascular endothelial cells in intestinal villi during early and advanced stages of lesions induced by C. perfringens type C. This was first associated with capillary dilatation and subsequently with widespread hemorrhage in affected intestinal segments. CPB was, however, not demonstrated on intestinal epithelial cells. This indicates a tropism of CPB toward endothelial cells and suggests that CPB-induced endothelial damage plays an important role in the early stages of C. perfringens type C enteritis in pigs.

Mo1781 Smoking is Associated With Disordered Homeostasis of Small Intestinal and Proximal Colonic Mucosal Mast Cells

Mo1781 Smoking is Associated With Disordered Homeostasis of Small Intestinal and Proximal Colonic Mucosal Mast Cells

Role of bone marrow-derived mesenchymal stem cells in a rat model of severe acute pancreatitis

To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells (MSCs) in severe acute peritonitis (SAP). Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups: non-sodium deoxycholate (SDOC) group (non-SODC group), SDOC group, and a MSCs intervention group (i.e., a co-culture system of MSCs and pancreatic acinar cells + SDOC). The cell survival rate, the concentration of malonaldehyde (MDA), the density of superoxide dismutase (SOD), serum amylase (AMS) secretion rate and lactate dehydrogenase (LDH) leakage rate were detected at various time points. In a separate study, Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group. Serum AMS, MDA and SOD, interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α levels, intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats. In both studies, the protective effect of MSCs was evaluated. In vitro, The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced, and the concentration of MDA, AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point. In vivo, Serum AMS, IL-6, TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group; however serum IL-10 level was higher than the SAP group. Serum SOD level was higher than the SAP group at each time point, whereas a significant between-group difference in SOD level was only noted after 24 h. Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs. MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium, promote the proliferation of enteric epithelium and repair of the mucosa, attenuate systemic inflammation in rats with SAP.

Traditional Herbal Medicine, Rikkunshito, Induces HSP60 and Enhances Cytoprotection of Small Intestinal Mucosal Cells as a Nontoxic Chaperone Inducer

Increasing incidence of small intestinal ulcers associated with nonsteroidal anti-inflammatory drugs (NSAIDs) has become a topic with recent advances of endoscopic technology. However, the pathogenesis and therapy are not fully understood. The aim of this study is to examine the effect of Rikkunshito (TJ-43), a traditional herbal medicine, on expression of HSP60 and cytoprotective ability in small intestinal cell line (IEC-6). Effect of TJ-43 on HSP60 expression in IEC-6 cells was evaluated by immunoblot analysis. The effect of TJ-43 on cytoprotective abilities of IEC-6 cells against hydrogen peroxide or indomethacin was studied by MTT assay, LDH-release assay, caspase-8 activity, and TUNEL. HSP60 was significantly induced by TJ-43. Cell necrosis and apoptosis were significantly suppressed in IEC-6 cells pretreated by TJ-43 with overexpression of HSP60. Our results suggested that HSP60 induced by TJ-43 might play an important role in protecting small intestinal epithelial cells from apoptosis and necrosis in vitro.

Overexpression of a 60-kDa heat shock protein enhances cytoprotective function of small intestinal epithelial cells

Aims With the advancement of small intestinal (double balloon and capsule) endoscopy technology, incidence of small intestinal lesion caused by nonsteroidal anti-inflammatory drugs (NSAIDs) has been known to be high. However, therapy for small intestinal mucosal lesion has not yet been developed. Previous studies have shown that heat shock proteins (HSPs) are involved in cytoprotection mediated by their function as a molecular chaperone. In this study, we examined the effect of HSP60 or HSP70 overexpression on hydrogen peroxide-induced (H 2O 2) or indomethacin-induced cell damage in the small intestinal epithelial cells. Main methods cDNA of human HSP60 or HSP70 was transfected to rat small intestinal (IEC-6) cells, and HSP60- or HSP70-overexpressing cells were cloned. IEC-6 cells transfected with vector only were used as control cells. These cells were treated with H 2O 2 (0–0.14 mM) or indomethacin (0–2.5 mM). The cell viability was determined by MTT-assay. Cell necrosis was evaluated by LDH-release assay. Further, apoptosis was evaluated by caspases-3/7 activity and TUNEL assay. Key findings Cell viability after H 2O 2 or indomethacin treatment was significantly higher in HSP60-overexpressing cells compared with that in control cells and HSP60-overexpressing cells. Apoptotic cells were also reduced in HSP60-overexpressing. Conclusion: These results indicate that HSP60 plays an important role in protecting small intestinal mucosal cells from H 2O 2-induced or indomethacin-induced cell injury. HSP70-overexpressing cells did not show anti-apoptotic ability. Significance These findings possibly suggest that function of each HSP is different in the small intestine. Therefore, for the therapy of small intestinal mucosal lesion, HSP60-induction therapy could be a new therapeutic strategy.

UPTAKE AND METABOLISM OF PLASMA GLUTAMINE BY THE SMALL INTESTINE

Abstract An isolated, vascularly perfused preparation of rat intestine extracted large amounts of glutamine (75 µmoles per hour), but no other amino acid, from a recirculated blood perfusate. With l-[U-14C]glutamine, the carbon was partly incorporated into tissue acid-insoluble material (14%) and the rest reappeared with little delay in intestinal venous blood in CO2 (57%), citrulline (6%), proline (5%), and organic acids (18%), predominantly citric acid and lactic acid. Perfusate glutamine was the source of 32% of the CO2 produced by the preparation, although the perfusate contained 10 to 15 mm glucose. Most glutamine uptake and metabolism occurred in the small intestinal mucosal cells and, from analyses of labeled tissue metabolites, appeared to proceed via glutamic acid. The rapid flow of glutamine carbon through mucosa may be explained in part by the small glutamine pool, only 0.2 µmole per g of tissue. Glutamine nitrogen taken up by intestine could be approximately accounted for by the quantities of citrulline (34%), alanine (33%), ammonia (23%), and proline (10%) released back into the perfusate. Glutamine uptake comparable to that found in the isolated organ was observed in vivo, from arteriovenous concentration measurements across intestine in fasted dogs, cats, hamsters, monkeys, and conventional and germ-free rats. Twenty-five to 33% of plasma glutamine was extracted in each pass through the tissue. Glutamine uptake persisted in rats during transport of large amounts of glutamic acid from the intestinal lumen into the blood and in rats injected with 6-diazo-5-oxo-l-norleucine. The results of these studies suggest that glutamine derived from blood is an important respiratory substrate for cells in the small intestinal mucosa and that intestine is a major site for metabolism of glutamine released into the circulation by other tissues.

The Lipid Messenger OEA Links Dietary Fat Intake to Satiety

The association between fat consumption and obesity underscores the need to identify physiological signals that control fat intake. Previous studies have shown that feeding stimulates small-intestinal mucosal cells to produce the lipid messenger oleoylethanolamide (OEA) which, when administered as a drug, decreases meal frequency by engaging peroxisome proliferator-activated receptors-alpha (PPAR-alpha). Here, we report that duodenal infusion of fat stimulates OEA mobilization in the proximal small intestine, whereas infusion of protein or carbohydrate does not. OEA production utilizes dietary oleic acid as a substrate and is disrupted in mutant mice lacking the membrane fatty-acid transporter CD36. Targeted disruption of CD36 or PPAR-alpha abrogates the satiety response induced by fat. The results suggest that activation of small-intestinal OEA mobilization, enabled by CD36-mediated uptake of dietary oleic acid, serves as a molecular sensor linking fat ingestion to satiety.

Adhesion of enterotoxigenic Escherichia coli in humans and animals.

Enterotoxigenic Escherichia coli (ETEC), an important cause of diarrhoea in humans and animal, require accessory virulence properties in addition to enterotoxin to manifest virulence. Several classes of pili (hair-like protein surface organelles) promote adhesion of ETEC to small intestinal mucosa. Antibody directed against adhesion pili interferes with colonization of the small intestine and prevents disease. This paper reviews studies with purified K88, K99 and 987 type pili used as parenteral vaccines in pregnant pigs and cattle. Infant animals suckled on immunized mothers were significantly protected against fatal disease. Colonization factor antigen (CFA) I and II pili, and type 1 somatic pili, promote adhesion of human ETEC pathogens to epithelial cells in vitro and are generally recognized as accessory virulence factors. CFA/I and II were found in only 25% of 36 human ETEC infections; positive strains were usually LT+/ST+ (LT: heat-labile; ST: heat-stable). Strains lacking CFA/I and II are virulent; other factors must be responsible for adhesion in such strains. While none of 14 LT+/ST- strains elaborated CFA/I or II, 10 (71%) possessed type 1 somatic pili. An initial ETEC diarrhoeal infection in volunteers stimulated protective immunity against diarrhoea on re-challenge with the same strain. Despite clinical protection healthy "veterans" excreted the ETEC strain to the same degree as ill controls. Thus the mechanisms of immunity was not bactericidal. Disease-induced LT antitoxic immunity failed to protect volunteers against challenge with a heterologous (LT+/ST-) strain. One explanation of these observations is that the mechanism of protection was anti-adhesive with antibody directed against adhesive factors on the bacterial surface preventing attachment of bacteria to receptors on small intestinal mucosal cells. Immunoprophylaxis against ETEC in humans with purified pili vaccines appears feasible.

Infiltration of forkhead box P3-expressing cells in small intestinal mucosa in coeliac disease but not in type 1 diabetes

Because the role of regulatory T cells in the intestinal inflammation is unknown in coeliac disease (CD) and type 1 diabetes (T1D), the expression of forkhead box P3 (FoxP3), CD25, transforming growth factor-beta, interferon (IFN)-gamma, interleukin (IL)-4, IL-8, IL-10, IL-15 and IL-18 was measured by quantitative reverse transcription-polymerase chain reaction in the small intestinal biopsies from paediatric patients with active or potential CD, T1D and control patients. The numbers of FoxP3- and CD25-expressing cells were studied with immunohistochemistry. Enhanced intestinal expressions of FoxP3, IL-10 and IFN-gamma mRNAs were found in active CD when compared with controls (P-values < 0.001, 0.004, <0.001). In potential CD, only the expression of IFN-gamma mRNA was increased. The numbers of FoxP3-expressing cells were higher in active and potential CD (P < 0.001, P = 0.05), and the ratio of FoxP3 mRNA to the number of FoxP3-positive cells was decreased in potential CD when compared with controls (P = 0.007). The ratio of IFN-gamma to FoxP3-specific mRNA was increased in active and potential CD (P = 0.001 and P = 0.002). Patients with T1D had no changes in regulatory T cell markers, but showed increased expression of IL-18 mRNA. The impaired up-regulation of FoxP3 transcripts despite the infiltration of FoxP3-positive cells in potential CD may contribute to the persistence of inflammation. The increased ratio of IFN-gamma to FoxP3 mRNA in active and potential CD suggests an imbalance between regulatory and effector mechanisms. The increased intestinal expression of IL-18 mRNA in patients with T1D adds evidence in favour of the hypothesis that T1D is associated with derangements in the gut immune system.