Rat Small Intestine Research Articles

Limoniastrum guyonianum prevents H2O2-induced oxidative damage in IEC-6 cells by enhancing enzyamtic defense, reducing glutathione depletion and JNK phosphorylation

Limoniastrum guyonianum is used in several regions of North Africa as a folk medicine. The objective of this study was to determine the in vitro antioxidant activities of L. guyonianum roots and their cytoprotective action on H2O2-challenged rat small intestine epithelial cells (IEC-6 cells). To assess the cytoprotective effect of L. guyonianum extract (LGE), IEC-6 cells were pre-incubated with different LGE concentrations. Then, IEC-6 cultures were exposed to 40μM H2O2 during 4h. Modulation of endogenous antioxidant system including SOD, CAT, MDA, GSH and the expression of possibly involved MAPKs was evaluated. Main results reported that L. guyonianum was rich in polyphenols and exhibited an important antioxidant activity as revealed by different tests (DPPH Assay, IC50=1.6μg/mL; ABTS+ test, IC50=27μg/mL; Fe-reducing power, EC50=44μg/mL). HPLC analysis showed that quercetin, catechin, and isorhamnetin-3-O-rutinoside were major phenolics. The exposure of IEC-6 cells to 40μM H2O2 during 4h resulted in oxidative stress manifested by (i) over 70% cell mortality, (ii) over-activity of CAT (246%), (iii) decrease in GSH level (10.4nmol/mg), (iv) excess in MDA content (18.4nmol/mg), and (v) a trigger of JNK phosphorylation. Pretreatment with LGE, especially at 0.25μg/mL, restored cell viability to 100%, and normal cell morphology in H2O2-chalenged cells. In addition, this extract maintained a high CAT activity, enhanced SOD capacity (120%) and increased GSH level (45.5nmol/mg). Furthermore, reducing cell death seems to be due to dephosphorylated JNK MAPK exerted by L. guyonianum bioactive compounds. In all, L. guyonianum components provided a cross-talk between regulatory pathways, implying their role as cytoprotector against oxidative stress.

Expression and regulation of the neutral amino acid transporter B0AT1 in rat small intestine.

Absorption of neutral amino acids across the luminal membrane of intestinal enterocytes is mediated by the broad neutral amino acid transporter B0AT1 (SLC6A19). Its intestinal expression depends on co-expression of the membrane-anchored peptidase angiotensin converting enzyme 2 (ACE2) and is additionally enhanced by aminopeptidase N (CD13). We investigated in this study the expression of B0AT1 and its auxiliary peptidases as well as its transport function along the rat small intestine. Additionally, we tested its possible short- and long-term regulation by dietary proteins and amino acids. We showed by immunofluorescence that B0AT1, ACE2 and CD13 co-localize on the luminal membrane of small intestinal villi and by Western blotting that their protein expression increases in distal direction. Furthermore, we observed an elevated transport activity of the neutral amino acid L-isoleucine during the nocturnal active phase compared to the inactive one. Gastric emptying was delayed by intragastric application of an amino acid cocktail but we observed no acute dietary regulation of B0AT1 protein expression and L-isoleucine transport. Investigation of the chronic dietary regulation of B0AT1, ACE2 and CD13 by different diets revealed an increased B0AT1 protein expression under amino acid-supplemented diet in the proximal section but not in the distal one and for ACE2 protein expression a reverse localization of the effect. Dietary regulation for CD13 protein expression was not as distinct as for the two other proteins. Ring uptake experiments showed a tendency for increased L-isoleucine uptake under amino acid-supplemented diet and in vivo L-isoleucine absorption was more efficient under high protein and amino acid-supplemented diet. Additionally, plasma levels of branched-chain amino acids were elevated under high protein and amino acid diet. Taken together, our experiments did not reveal an acute amino acid-induced regulation of B0AT1 but revealed a chronic dietary adaptation mainly restricted to the proximal segment of the small intestine.

Quercetin attenuates the ischemia reperfusion induced COX-2 and MPO expression in the small intestine mucosa

Quercetin, the active substance of tea, fruits and vegetables, exerts a broad spectrum of pharmacological activities and is considered to have potential therapeutic application. The present study was designed to investigate the beneficial effect of quercetin against experimental ischemia- reperfusion (IR) injury of the small intestine in rats. Quercetin was administrated intraperitoneally 30min before 1h ischemia of superior mesenteric artery with following reperfusion periods lasting 1, 4 and 24h. The male specific pathogen-free Charles River Wistar rats were used (n=45). In acute phase, 4h after start of reperfusion, the quercetin induced a significant decrease in mucosal injury index (p<0.05) accompanied by a significant decrease in cyclooxygenase-2 (COX-2) expression in the epithelial lining of the intestinal villi in comparison with the control group (p<0.01). In the epithelium of the intestinal glands, COX-2 expression resulting from IR injury significantly increased regardless quercetin application (in control group p<0.001; in quercetin group p<0.05), but in quercetin group, significant decrease in it during 24h of reperfusion in a late phase of IR injury was detected (p<0.001). Based on morphology of COX-2 positive cells, the COX-2 positivity was found particularly in goblet cells of the intestinal villi epithelium and enteroendocrine cells respectively, in the glandular epithelium. We concluded that quercetin application attenuated mucosal damage from IR injury by inhibiting neutrophil infiltration which was demonstrated by a lower number of myeloperoxidase positive cells in the lamina propria during both phases of IR injury and the significant decrease in that in a late phase after 24h of reperfusion (p<0.05).

MiR-146a protects small intestine against ischemia/reperfusion injury by down-regulating TLR4/TRAF6/NF-κB pathway.

Previous studies reported that miR-146a was involved in small intestine ischemia-reperfusion (I/R) injury, but the mechanism is largely vague. Here, we aimed to identify the change of miR-146a in patients with mesenteric ischemia and explore the potential regulatory mechanism of miR-146a in intestine epithelial cells survival under ischemia and I/R injury. The plasma of 20 patients with mesenteric ischemia and 25 controls was collected to examine the miR-146a expression by qPCR. Rat intestinal epithelial cells (IEC-6) and 24 male Sprague-Dawley rats were included to build ischemia and I/R model in vitro and in vivo. The qPCR results showed that miR-146a decreased both in the plasma of patients with mesenteric ischemia and in IEC-6 cells and rat small intestine tissues in ischemia and I/R model compared to controls. Both the in vitro and in vivo results showed that I/R resulted in more severe apoptotic injury than ischemia. Cleaved-caspase 3, TLR4, TRAF6, and nuclear NF-κB p65 were up-regulated accompanying reduced XIAP and SOCS3 expression in intestinal ischemia and I/R injury. After up-regulation of miR-146a in IEC-6 cells, increased cell survival and decreased cell apoptosis were observed, concomitant with decreased cleaved-caspase 3 and down-regulated TLR4/TRAF6/NF-κB pathway. What is more, this protective effect was blocked by TRAF6 overexpression and increased nuclear NF-κB p65 nuclear. Taken together, this study revealed that miR-146a expression was decreased in small intestine ischemia and I/R injury. And miR-146a improves intestine epithelial cells survival under ischemia and I/R injury through inhibition TLR4, TRAF6, and p-IκBα, subsequently leading to decreased NF-κB p65 nuclear translocation.

Effects of 1,5-anhydroglucitol on postprandial blood glucose and insulin levels and hydrogen excretion in rats and healthy humans.

The inhibition by 1,5-anhydro-d-glucitol (1,5-AG) was determined on disaccharidases of rats and humans. Then, the metabolism and fate of 1,5-AG was investigated in rats and humans. Although 1,5-AG inhibited about 50 % of sucrase activity in rat small intestine, the inhibition was less than half of d-sorbose. 1,5-AG strongly inhibited trehalase and lactase, whereas d-sorbose inhibited them very weakly. 1,5-AG noncompetitively inhibited sucrase. The inhibition of 1,5-AG on sucrase and maltase was similar between humans and rats. 1,5-AG in serum increased 30 min after oral administration of 1,5-AG (600 mg) in rats, and mostly 100 % of 1,5-AG was excreted into the urine 24 h after administration. 1,5-AG in serum showed a peak 30 min after ingestion of 1,5-AG (20 g) by healthy subjects, and decreased gradually over 180 min. About 60 % of 1,5-AG was excreted into the urine for 9 h following ingestion. Hydrogen was scarcely excreted in both rats and humans 24 h after administration of 1,5-AG. Furthermore, 1,5-AG significantly suppressed the blood glucose elevation, and hydrogen excretion was increased following the simultaneous ingestion of sucrose and 1,5-AG in healthy subjects. 1,5-AG also significantly suppressed the blood glucose elevation following the simultaneous ingestion of glucose and 1,5-AG; however, hydrogen excretion was negligible. The available energy of 1,5-AG, which is absorbed readily from the small intestine and excreted quickly into the urine, is 0 kJ/g (0 kcal/g). Furthermore, 1,5-AG might suppress the blood glucose elevation through the inhibition of sucrase, as well as intestinal glucose absorption.

Inhibitory effect of rhubarb on intestinal α-glucosidase activity in type 1 diabetic rats

Purpose: To investigate the inhibitory effect of rhubarb on α-glucosidase activity in the small intestine of rats with type 1 diabetes.Methods: Type 1 diabetic rat model was established by intraperitoneally injecting 30 male SD rats with 1 % streptozocin (STZ). Rats with fasting blood glucose &gt; 11 mmol/L (24) were used for the study. The rats were randomly divided into three equal groups including control, acarbose and rhubarb groups. Arcabose® (20 mg/kg /day) and rhubarb (100 mg/kg /day) were given by intra-gastric route via insertion of the cannula through the esophagus. Daily fasting blood glucose and daily postprandial glucose levels were assayed for all groups. On day 6, postprandial blood glucose, blood levels of C-peptide and insulin, and intestinal α-glucosidase were also determined.Results: There were no significant differences in levels of C-peptide, insulin and fasting blood glucose between control, Acarbose® and rhubarb groups (p &gt; 0.05). However, α-glucosidase activity at 0, 30, 60 and 120 min in the rhubarb group was 1759.2, 1812.8, 1379.8 and 772.1 U, respectively,) while in the Acarbose® group it was 178.6, 1260.1, 1126.5, 599.2 U, respectively. α-Glucosidase activity in both groups initially showed an increase (p &lt; 0.05), followed by a decline from 60 to 120 min (p ˂ 0.05). After 120 min, α-glucosidase activity in each of the two groups was significantly decreased compared with untreated control (1200 U) (p ˂ 0.05).Conclusion: The inhibitory effect of rhubarb on intestinal α-glucosidase activity of Type 1 diabetic rats is comparable to that of Arcabose®.This suggests that this plant may have clinically potent anti-diabetic properties.Keywords: Type 1 diabetes, α-Glucosidase activity, Acarbose®, Rhubarb, Postprandial glucose level

Erratum to: Synergistic effect of aluminum and ionizing radiation upon ultrastructure, oxidative stress and apoptotic alterations in Paneth cells of rat intestine

Environmental and occupational exposure to aluminum along with ionizing radiation results in serious health problems. This study was planned to investigate the impact of oxidative stress provoked by exposure to ionizing radiation with aluminum administration upon cellular ultra structure and apoptotic changes in Paneth cells of rat small intestine . Animals received daily aluminum chloride by gastric gavage at a dose 0.5 mg/Kg BW for 4 weeks. Whole body gamma irradiation was applied at a dose 2 Gy/week up to 8 Gy. Ileum malondialdehyde, advanced oxidation protein products, and protein carbonyl were assessed as biomarkers of lipid peroxidation along with superoxide dismutase, catalase, and glutathione peroxidase activities as enzyme antioxidants. Moreover, analyses of cell cycle division and apoptotic changes were evaluated by flow cytometry. Intestinal cellular ultra structure was investigated using transmission electron microscope. Oxidative stress assessment in the ileum of rats revealed that aluminum and ionizing radiation exposure either alone or in combination exhibits a significant effect upon the increase in biomarkers of lipid peroxidation along with tumor necrosis factor-α with concomitant significant decrease of the antioxidant enzyme activities. Flow cytometric analyses showed significant alterations in the percentage of cells during cell cycle division phases along with significant increase in apoptotic cells. Ultra structurally, intestinal cellular alterations with marked injury in Paneth cells at the sites of bacterial translocation in the crypt of lumens were recorded. The results of this study have clearly suggested that aluminum exposure and ionizing either alone or in combination induced apoptosis and oxidative stress in the Paneth cells of rat intestine, which appeared to play a major role in the pathogenesis of cellular damage. Furthermore, the interaction of these two intestinal toxic routes was found to be synergistic.

Lactadherin orthologs inhibit migration of human, porcine and murine intestinal epithelial cells.

Lactadherin was originally described due to its appearance in milk, but is abundantly expressed especially by professional and nonprofessional phagocytes. The proteins has been shown to have a multitude of bioactive effects, including inhibition of inflammatory phospholipases, induction of effero‐ and phagocytosis, prevent rotavirus induced gastroenteritis, and modulate intestinal homeostasis by regulating epithelial cell migration. The level of expression seems to be important in a row of serious pathologies linked to the intestinal epithelial barrier function, vascular‐ and autoimmune disease. This study examines the ability of lactadherin to modulate migration of intestinal epithelium. A cell exclusion assay is used to quantify the ability of human, bovine and murine lactadherin orthologs to affect migration of primary small intestine epithelium cells. Previous reports show that recombinant murine lactadherin stimulate rat small intestine cell migration. The present study could not confirm this. Conversely, 10 μg/ml lactadherin inhibits migration. Therefore, as lactadherins enteroprotective properties is well established using in vivo models we conclude that the protective effects are linked to lactadherins ability operate as an opsonin, or other modulating effects, and not a direct lactadherin‐cell induction of migration. Thus, the molecular mechanism behind the enteroprotective role of lactadherin remains to be established.

Intestinal absorption mechanisms of 2′-deoxy-2′-β-fluoro-4′-azidocytidine, a cytidine analog for AIDS treatment, and its interaction with P-glycoprotein, multidrug resistance-associated protein 2 and breast cancer resistance protein

2′-Deoxy-2′-β-fluoro-4′-azidocytidine (FNC), a cytidine analog, has attracted great interest because of its potent activity against wild-type and multidrug-resistant HIV. The purpose of current study was to investigate the absorption mechanisms of FNC in the small intestine, as well as the interactions between FNC and P-glycoprotein (P-gp), multidrug resistance-associated protein 2 (MRP2) and breast cancer resistance protein (BCRP). The experiments were performed using Caco-2 cells and the rat small intestine. The uptake experiment indicated that FNC concentration, extracellular pH and the incubated temperature could influence the uptake of FNC in Caco-2 cells. NaN3, verapamil, probenecid, MK571 and GF120918 could significantly increase the FNC uptake in Caco-2 cells. The transport experiment showed that both the absorption and secretion of FNC were concentration dependent. The secretion of FNC was approximately 2-fold greater than the absorption. In the presence of verapamil, probenecid, MK571 or GF120918, the efflux ratio decreased by >50%. In everted rat intestine, the absorption of FNC also depended on its concentration and was not significantly different in the different segments of the small intestine. Real-time RT-PCR results indicated that the gene expressions of P-gp, MRP2 and BCRP were up-regulated after exposure to FNC. The reduction in accumulation of rhodamine 123 after treatment with FNC revealed its ability to up-regulate P-gp activity. In conclusion, FNC was completely absorbed by passive diffusion and active transport mechanisms. P-gp, MRP2 and BCRP could influence the absorption of FNC in the small intestine. FNC could modulate the gene expressions of P-gp, MRP2 and BCRP, and increase the activity of P-gp.

Assessment of absorption of four lignan constituents of JingNing particles in rat gut using &lt;i&gt;in situ&lt;/i&gt; single-pass intestinal perfusion

Purpose: To study small intestinal absorption of schisadrol A, schisandrol B, schizandrin A and schisandrin B in JingNing particles using in situ single-pass intestinal perfusion (SPIP).Methods: Absorption rate constant (Ka) and apparent permeability (Papp) of the drugs at different concentrations in various parts of rat small intestine (duodenum, jejunum and ileum) were determined using SPIP. JingNing particles were also perfused in situ at different pH in the entire rat intestine. Ethanol extract of Schisandra chinensis (standard) at low concentration was perfused in the duodenum for comparison with extract of JingNing particles.Results: The order of apparent permeability of the four lignans was schisandrol A &lt; schisandrol B &lt; schizandrin A &lt; schisandrin B. Ka and Papp values of the four lignans in JingNing particles were concentration-dependent. Absorption increased in the rank order: ileum &gt; duodenum &gt; jejunum. Optimum absorption pH was 6.50. Polygala tenuifolia extract and volatile oil of Rhizoma acori tatarinowii significantly (p &lt; 0.05) enhanced the absorption of the four lignans.Conclusion: The four lignans were well absorbed in the intestinal tract, particularly the ileum, probably through carrier-mediated transport. The alcohol extract of Polygala tenuifolia and volatile oil of Rhizoma acori graminei enhanced the absorption of the four lignans.Keywords: JingNing, Intestinal absorption, Polygala tenuifolia, Rhizoma acori graminei, Lignans, Schisandrol, Schisandrin, Single-pass intestinal perfusion

Sweet Taste Receptor Activation in the Gut Is of Limited Importance for Glucose-Stimulated GLP-1 and GIP Secretion.

Glucose stimulates the secretion of the incretin hormones: glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP). It is debated whether the sweet taste receptor (STR) triggers this secretion. We investigated the role of STR activation for glucose-stimulated incretin secretion from an isolated perfused rat small intestine and whether selective STR activation by artificial sweeteners stimulates secretion. Intra-luminal administration of the STR agonists, acesulfame K (3.85% w/v), but not sucralose (1.25% w/v) and stevioside (2.5% w/v), stimulated GLP-1 secretion (acesulfame K: 31 ± 3 pmol/L vs. 21 ± 2 pmol/L, p < 0.05, n = 6). In contrast, intra-arterial administration of sucralose (10 mM) and stevioside (10 mM), but not acesulfame K, stimulated GLP-1 secretion (sucralose: 51 ± 6 pmol/L vs. 34 ± 4 pmol/L, p < 0.05; stevioside: 54 ± 6 pmol/L vs. 32 ± 2 pmol/L, p < 0.05, n = 6), while 0.1 mM and 1 mM sucralose did not affect the secretion. Luminal glucose (20% w/v) doubled GLP-1 and GIP secretion, but basolateral STR inhibition by gurmarin (2.5 µg/mL) or the inhibition of the transient receptor potential cation channel 5 (TRPM5) by triphenylphosphine oxide (TPPO) (100 µM) did not attenuate the responses. In conclusion, STR activation does not drive GIP/GLP-1 secretion itself, nor does it have a role for glucose-stimulated GLP-1 or GIP secretion.

Melatonin protects rats from radiotherapy-induced small intestine toxicity.

Radiotherapy-induced gut toxicity is among the most prevalent dose-limiting toxicities following radiotherapy. Prevention of radiation enteropathy requires protection of the small intestine. However, despite the prevalence and burden of this pathology, there are currently no effective treatments for radiotherapy-induced gut toxicity, and this pathology remains unclear. The present study aimed to investigate the changes induced in the rat small intestine after external irradiation of the tongue, and to explore the potential radio-protective effects of melatonin gel. Male Wistar rats were subjected to irradiation of their tongues with an X-Ray YXLON Y.Tu 320-D03 irradiator, receiving a dose of 7.5 Gy/day for 5 days. For 21 days post-irradiation, rats were treated with 45 mg/day melatonin gel or vehicle, by local application into their mouths. Our results showed that mitochondrial oxidative stress, bioenergetic impairment, and subsequent NLRP3 inflammasome activation were involved in the development of radiotherapy-induced gut toxicity. Oral treatment with melatonin gel had a protective effect in the small intestine, which was associated with mitochondrial protection and, consequently, with a reduced inflammatory response, blunting the NF-κB/NLRP3 inflammasome signaling activation. Thus, rats treated with melatonin gel showed reduced intestinal apoptosis, relieving mucosal dysfunction and facilitating intestinal mucosa recovery. Our findings suggest that oral treatment with melatonin gel may be a potential preventive therapy for radiotherapy-induced gut toxicity in cancer patients.

Short N-terminal galanin fragments are occurring naturally in vivo

The galanin family currently consists of four peptides, namely galanin, galanin-message associated peptide, galanin-like peptide and alarin. Unlike galanin that signals through three different G protein-coupled receptors; GAL1, GAL2, and GAL3, binding at its N-terminal end, the cognate receptors for other members of the galanin family are currently unknown. Research using short N-terminal galanin fragments generated either by enzymatic cleavage or solid-phase synthesis has revealed differences in their receptor binding properties exerting numerous biological effects distinct from galanin(1-29) itself. Our studies on tissue extracts derived from rat small intestine and bovine gut using chromatographic techniques and sensitive galanin(1-16)-specific radioimmunoassay revealed the presence of immunoreactive compounds reacting with antiserum against galanin(1-16) distributed in distinct elution volumes. These results suggested a possible presence of short N-terminal galanin fragments also in vivo. Moreover, employing immunoaffinity chromatography and reverse-phase high performance liquid chromatography (HPLC) followed by mass spectrometry allowed specific enrichment of these immunoreactive compounds from rat tissues and identification of their molecular structure. Indeed, our study revealed presence of several distinct short N-terminal galanin sequences in rat tissue. To prove their receptor binding, four of the identified sequences were synthetized, namely, galanin(1-13), galanin(1-16), galanin(1-20), galanin(6-20), and tested on coronal rat brain sections competing with 125I-labeled galanin(1-29). Our autoradiographs confirmed that galanin(1-13), galanin(1-16), and galanin(1-20) comprehensively displaced 125I-galanin(1-29) but galanin(6-20) did not. Here we show, for the first time, that short N-terminal galanin fragments occur naturally in rat tissues and that similar or identical galanin sequences can be present also in tissues of other species. Biological significanceThis study is first to provide an evidence of the presence of short N-terminal galanin fragments in vivo in a biological system and provides further foundations for the previous studies using synthetized short N-terminal galanin fragments.

Rapid LPS transport during fat absorption in rat small intestine

Lipopolysaccharide (LPS; endotoxin) is a lipophilic pathogenic factor derived from gut Gram‐negative bacteria. Increased serum LPS concentrations are associated with the metabolic syndrome, termed metabolic endotoxemia. Since the origin of circulating LPS is the gut lumen, we studied the mechanisms of LPS transport across the gut mucosa, hypothesizing that transport occurs via known lipid uptake pathways. Since glucagon‐like peptide‐2 (GLP‐2) reduces intestinal solute permeability but enhances fat absorption, we also hypothesized that GLP‐2 affects LPS transport during fat absorption. We thus examined LPS transport mechanisms and the effect of exogenous GLP‐2 on LPS transport in intact intestinal tissues in vitro and in vivo. FITC‐LPS absorption in vivo was quantitated by simultaneously measuring the appearance of FITC‐LPS into the portal vein (PV) and the mesenteric lymph, with or without luminal oleic acid with taurocholate (OA/TCA). We also examined fluorescent dextran 4000 (FD4) or LPS transport from mucosal to serosal solution (m‐to‐s) in Ussing chambered muscle‐stripped intestinal tissues.Bolus intraduodenal infusion of FITC‐LPS (50 μg/ml) with OA/TCA rapidly increased FITC‐LPS appearance into PV, followed by a gradual increase into the lymph, whereas FITC‐LPS alone did not appear on PV or lymph. In Ussing chambered jejunum, luminally‐applied LPS (10 μg/ml) rapidly appeared in the serosal solution with luminal OA/TCA, whereas LPS alone or LPS+TCA was not transported. OA/TCA‐induced LPS m‐to‐s transport in the jejunum was inhibited by luminal pretreatment with the lipid raft inhibitor methyl‐β‐cyclodextrin and the CD36 inhibitor sulfosuccinimidyl oleate, but was enhanced by serosal carbachol and luminal application of the competitive alkaline phosphatase inhibitor glycerol phosphate. Furthermore, FD4 m‐to‐s movement was also enhanced by luminal OA/TCA in the jejunum, whereas lesser effects were observed in the duodenum or ileum. Pretreatment with serosal GLP‐2 (100 nM) reduced OA/TCA‐induced LPS transport, whereas carbachol‐induced LPS transport was not affected.These results suggest that luminal LPS crosses the gut barrier physiologically during fat absorption via lipid raft‐ and CD36‐mediated transport mechanisms, similar to chylomicron transport, and possibly through emptied goblet cells. By reducing lipid raft‐, CD36‐, and chylomicron‐mediated LPS transport, GLP‐2 treatment may be a new therapy for metabolic endotoxemia.Support or Funding InformationSupported by VA Merit Review, NIH R01 DK54221

1087 - Rapid LPS Transport During Long-Chain Fatty Acid Absorption in Rat Small Intestine

1087 - Rapid LPS Transport During Long-Chain Fatty Acid Absorption in Rat Small Intestine

The time changes of endogenous salusin-β in septic rats

Salusin-α and salusin-β are newly found bioactive peptides of 28 and 20 amino acids, respectively, which are widely distributed in the hematopoietic system, endocrine system, and central nervous system. Salusins exert cardiovascular effects, including hypotension and bradycardia; promote vascular inflammation; and so on. However, little information is available yet on the relationships of salusin-β with sepsis. In this study, we investigated the distribution and content of endogenous salusin-β in septic rats. A total of 72 specified pathogen-free (SPF) male Sprague-Dawley (SD) rats were randomly divided into control group (sham operation, n = 36) and experimental group (n = 36) with sepsis rat model by cecal ligation and puncture (CLP). The model group rats were sacrificed after 6, 12, and 24 h of treatment. The concentration of salusin-β in spleen, stomach, small intestine, hypothalamus, and serum specimens was detected by enzyme-linked immunosorbent assay (ELISA). It showed that salusin-β was endogenously generated in rat tissues, including spleen, stomach, small intestine, hypothalamus, and serum. The content of salusin-β in the spleen was higher than that in other tissues. The content of salusin-β in the spleen, stomach, and small intestine, together with the serum level of salusin-β, increased significantly at 6 h after CLP compared with the control group ( P &lt; 0.05). The content of salusin-β in spleen and serum peaked at 12 h, and in small intestine, it reached the summit at 24 h. Meanwhile, no significant fluctuations in salusin-β content were observed in the stomach. The content of salusin-β in hypothalamus began to increase at 6 h, and a significant increase appeared 12 h after CLP ( P &lt; 0.05). In conclusion, this study shows that the time-dependent alterations of salusin-β in septic rats suggest that salusin-β might be involved in the pathogenesis of sepsis.

Pharmacokinetic changes of norfloxacin based on expression of MRP2 after acute exposure to high altitude at 4300m.

This study was to investigate the influence of physiological changes and the expression of MRP2 efflux transporter on the pharmacokinetics of norfloxacin after acute exposure to high altitude 4300m. The rats were randomly divided into high altitude group and plain group. Blood gas and biochemical analysis showed that the physiological parameters significantly changed at high altitude. The mRNA and protein expression of MRP2 in high altitude group were higher than plain group in rat small intestine and kidney, while was reduced in rat liver. The AUC, Ka and Cmax of norfloxacin were significantly reduced in high altitude group (p<0.05). However, the MRT, CL, t1/2 and Vd were significantly increased (p<0.05). These results indicate that physiological indicators and expression levels of drug transporters MRP2 are changed in responded to high altitude, to severely affect norfloxacin pharmacokinetics. These changes may provide basis and new ideas to adjust the dosage and administration, so as to promote rational drug use in the high altitude.

Biochemical and histological changes in female wistar rats following oral administration of aqueous extract of &lt;i&gt;Mangifera indica&lt;/i&gt; L. leaves

This study was aimed at investigating the effect of aqueous extract of Mangifera indica leaves on some biochemical parameters and histology of liver, kidney and small intestine of rats. Twenty female rats (142.30 ± 7.56 g) were randomly assigned into four groups A, B, C and D. The rats were administered orally 1 mL of distilled water, 25, 50 and 100 mg/kg body weight of the extract once daily for 10 days. The biochemical parameters were determined using standard methods. All the doses of the extract significantly increased the activities of liver aspartate aminotransferase (AST), serum alanine aminotransferase (ALT) and chloride ion (Cl - ) content whereas the liver ALT, serum urea, uric acid, potassium ion (K + ) and small intestine body weight ratio decreased significantly in a dose dependent manner. The serum concentrations of sodium ion (Na + ) and creatinine increased only at 50 and 100 mg/kg body weight of the extract while the levels of total protein, albumin, globulin, conjugated and total bilirubin significantly decreased in the serum. The liver and kidney body weight ratio significantly increased at 100 mg/kg body weight of the extract while the extract at all the doses did not significantly alter the activity of serum AST. Furthermore, the extract significantly decreased the activity of alkaline phosphatase (ALP) in the small intestine of the animals with corresponding increase in the serum. The extract at varying doses produced mild and moderate hepatocellular and submucosa layer degeneration in the liver and small intestine respectively while there was glomeruli shrinkage in the kidney. The extract caused structural and functional toxicity hence, should be consumed with caution. Keywords: Mangifera indica , Anacardiaceae, Structural toxicity, Functional toxicity, Histoarchitectural changes

Biochemical and histological changes in female wistar rats following oral administration of aqueous extract of &lt;i&gt;Mangifera indica&lt;/i&gt; L. leaves

This study was aimed at investigating the effect of aqueous extract of Mangifera indica leaves on some biochemical parameters and histology of liver, kidney and small intestine of rats. Twenty female rats (142.30 ± 7.56 g) were randomly assigned into four groups A, B, C and D. The rats were administered orally 1 mL of distilled water, 25, 50 and 100 mg/kg body weight of the extract once daily for 10 days. The biochemical parameters were determined using standard methods. All the doses of the extract significantly increased the activities of liver aspartate aminotransferase (AST), serum alanine aminotransferase (ALT) and chloride ion (Cl-) content whereas the liver ALT, serum urea, uric acid, potassium ion (K+) and small intestine body weight ratio decreased significantly in a dose dependent manner. The serum concentrations of sodium ion (Na+) and creatinine increased only at 50 and 100 mg/kg body weight of the extract while the levels of total protein, albumin, globulin, conjugated and total bilirubin significantly decreased in the serum. The liver and kidney body weight ratio significantly increased at 100 mg/kg body weight of the extract while the extract at all the doses did not significantly alter the activity of serum AST. Furthermore, the extract significantly decreased the activity of alkaline phosphatase (ALP) in the small intestine of the animals with corresponding increase in the serum. The extract at varying doses produced mild and moderate hepatocellular and submucosa layer degeneration in the liver and small intestine respectively while there was glomeruli shrinkage in the kidney. The extract caused structural and functional toxicity hence, should be consumed with caution.Keywords: Mangifera indica, Anacardiaceae, Structural toxicity, Functional toxicity, Histoarchitectural changes

Vascular Pathology of Ischemia/Reperfusion Injury of Rat Small Intestine

Ischemia/reperfusion (I/R) injury of the small intestine caused by occlusion of the superior mesenteric artery affects the intestinal tissue as well as components of the blood circulatory system from the microvasculature to mesenteric vessels. The aim of this work was to study the correlation between the dynamics of destruction development in the intestinal tissue, microvasculature, and mesenteric vessels in I/R of the small intestine. The microvasculature was analyzed by whole-organ continuous monitoring of the intestinal mucosal blood perfusion by laser Doppler flowmetry during the entire I/R. Real-time RT-PCR was used to assess gene expression of NF-κB, caspase-3, Ki67, and TNF-α in blood vessels. At the start of reperfusion, the first targets to be disrupted are microvessels in the apical villi. Injury of the apical part of the microcirculatory bloodstream correlates with the reduction in intestinal mucosal blood perfusion, which occurred simultaneously with apical villous destruction. By the end of the reperfusion period, the low intestinal mucosal blood perfusion is mirrored by the destruction of the microvasculature and mucosal structures in the entire organ. The development of mesenteric vessel injury is characterized by a change in NO metabolism and damaged endothelial cells concomitant with an alteration in the expression of genes encoding NF-κB, caspase-3, and Ki67 by the end of the reperfusion period. In I/R injury, detrimental effects on the intestinal tissue, microvasculature, and mesenteric vessels develop and exhibit common mechanisms of function, which show strong correlations.