Descending Colon Mucosa Research Articles

Structure-Activity Relationship Study of the High-Affinity Neuropeptide Y4 Receptor Positive Allosteric Modulator VU0506013.

Positive allosteric modulators targeting the Y4 receptor (Y4R), a G protein-coupled receptor (GPCR) involved in the regulation of satiety, offer great potential in anti-obesity research. In this study, we selected 603 compounds by using quantitative structure-activity relationship (QSAR) models and tested them in high-throughput screening (HTS). Here, the novel positive allosteric modulator (PAM) VU0506013 was identified, which exhibits nanomolar affinity and pronounced selectivity toward the Y4R in engineered cell lines and mouse descending colon mucosa natively expressing the Y4R. Based on this lead structure, we conducted a systematic SAR study in two regions of the scaffold and presented a series of 27 analogues with modifications in the N- and C-terminal heterocycles of the molecule to obtain insight into functionally relevant positions. By mutagenesis and computational docking, we present a potential binding mode of VU0506013 in the transmembrane core of the Y4R. VU0506013 presents a promising scaffold for developing in vivo tools to move toward anti-obesity drug research focused on the Y4R.

Identification and Characterization of the First Selective Y4 Receptor Positive Allosteric Modulator.

The human Y4 receptor (Y4R) and its cognate ligand, pancreatic polypeptide (PP), are involved in the regulation of energy expenditure, satiety, and food intake. This system represents a potential target for the treatment of metabolic diseases and has been extensively investigated and validated in vivo. Here, we present the compound tBPC (tert-butylphenoxycyclohexanol), a novel and selective Y4R positive allosteric modulator that potentiates Y4R activation in G-protein signaling and arrestin3 recruitment experiments. The compound has no effect on the binding of the orthosteric ligands, implying its allosteric mode of action at the Y4R and evidence for a purely efficacy-driven positive allosteric modulation. Finally, the ability of tBPC to selectively potentiate Y4R agonism initiated by PP was confirmed in mouse descending colon mucosa preparations expressing native Y4R, demonstrating Y4R positive allosteric modulation in vitro.

Antiphospholipid Syndrome Presenting as Thrombotic Acute Mesenteric Ischemia of an Anomalous Inferior Mesenteric Vein

Introduction: Thrombosis of the splanchnic circulationis an uncommon manifestation of Antiphospholipid Syndrome (APS); a rare site of which is the Inferior Mesenteric Vein (IMV). The rarest etiology of Acute Mesenteric Ischemia (AMI) is thrombosis of the venous supply (2-5%). We present a case of a patient found to have APS presenting with AMI of the left colon secondary to thrombosis of an anomalous IMV. Case Report: A 76 y/o female presented with bright red blood per rectum, mild abdominal pain and tenderness with no rigidity or guarding. Investigations revealed nonspecific rectal and distal sigmoid inflammatory process on non-contrast CT. A flexible sigmoidoscopy revealed multiple ulcers in the rectum and sigmoid colon, with congested mucosa and exudates. Proximally the descending colon mucosa gradually normalized in appearance with a clear marginal zone which was suspicious for an ischemic etiology (Figure 1). An abdominal angiogram revealed multiple filling defects in a dilated IMV abnormally draining into the Superior Mesenteric Vein (SMV) with significant wall thickening of the left colon (Figure 2). Intravenous heparin drip was initiated. A rapid improvement in symptoms was observed. An anti-phospholipid antibody panel showed an increase in IgA anti β2- glycoprotein, confirming APS. The patient was bridged to Rivaroxiban. A re-evaluation after six months revealed no recurrence of symptoms. A follow up Colonoscopy showed normal mucosa of the colon with complete resolution of mucosal ischemic changes (Figure 3).Figure 1Figure 2Figure 3Discussion: APS presented itself with thrombosis in the least common location within the splanchnic circulation in our patient. The most common site of all- cause splanchnic thrombosis is the SMV (50 to 82%) and rarely involves the IMV (14%). Our patient has an anomaly by which the IMV drains into the SMV (normally drains into the splenic vein) which may explain the rarity of the location of her thrombosis and left colonic pathology. The early detection of our patient's thrombosis and early intervention with anticoagulation therapy was essential in avoiding bowel wall necrosis and the need for surgical intervention. Our experience with managing this patient highlights the benefits of maintaining a high level of suspicion for thrombosis of the splanchnic circulation in patients presenting with abdominal pain and blood per rectum.

S1615 Phenylthiocarbamide Modulates Ion Secretion via Release of PYY and 5-HT in Mouse Descending Colon Mucosa

S1615 Phenylthiocarbamide Modulates Ion Secretion via Release of PYY and 5-HT in Mouse Descending Colon Mucosa

Expression of aquaporin 8 in colonic epithelium with diarrhoea-predominant irritable bowel syndrome

We analysed and compared the aquaporin 8 (AQP8) expression in ascending and descending colon mucosa between patients with diarrhoea-predominant irritable bowel syndrome (D-IBS) and healthy volunteers, in order to study the relationship between the clinical feature of IBS, the expression of AQP8 and the pathological mechanism of D-IBS. Specimens were taken from the proximal ascending colon or distal descending colon of D-IBS patients (n = 26) and healthy volunteers (n = 30), and AQP8 mRNA expression of each specimen was determined by fluorescent quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR). In patients with D-IBS, the relationship was analysed between AQP8 expression in both ascending and descending colons and clinical features including gender, age of onset, duration of illness, frequency of defecation, and stool characteristics. Although AQP8 was present in the epithelium of the ascending and descending colons in healthy persons and D-IBS patients, the AQP8 level of the D-IBS patients was significantly lower than that of the healthy persons (P < 0.01 in the ascending colon, P < 0.05 in the descending colon). AQP8 expression was not correlated with the age of patients with D-IBS (P > 0.05 both in the ascending and descending colons) or the age at the onset (P > 0.05 both in the ascending and descending colons), but closely with the duration of illness (P < 0.05 in the ascending colon, P < 0.01 in the descending colon), frequency of defecation (P < 0.01 in the ascending colon, P < 0.05 in the descending colon) and stool characteristics (P < 0.01 in the ascending colon, P > 0.05 in the descending colon). The decreased AQP8 expression in D-IBS patients indicates that dysfunction of colonic absorption may cause reduced water absorption, loose stool and diarrhoea. The expression of AQP8 may be related to D-IBS.

Receptors for calcitonin gene related peptide (CGRP) in gastrointestinal epithelia

A pharmacological comparison of calcitonin gene related peptide (CGRP) receptors expressed in normal rat colon mucosa and two human adenocarcinoma cell lines has been undertaken. Using voltage-clamp techniques electrogenic ion transport was continuously monitored across either mucosal preparations or confluent epithelial monolayers grown on permeable supports. The data presented at this meeting show that CGRP receptors are preferentially located on the basolateral epithelial surface and that their stimulation by a variety of CGRP analogues results in enhanced CI- secretion mediated via a cyclic AMP dependent mechanism. Responses to rat alpha CGRP in rat descending colon mucosa and in the adenocarcinoma cell line Colony-29 are insensitive to the inhibitory effects of the C-terminal fragment human CGRP(8-37); however, significant inhibition of rat alpha CGRP responses was observed in the parent epithelial cell line HCA-7. This together with the subtle differences seen in agonist orders of potency in the three preparations indicates that different CGRP receptor subtypes exist in the basolateral domains of HCA-7 compared with rat colon and Colony-29 epithelia.

Pharmacological characterisation of neurokinin receptors mediating anion secretion in rat descending colon mucosa

Substance P (SP), neurokinin A (NKA), neurokinin B (NKB), [Sar9, Met (O2)11]-SP (SMSP), senktide, [beta Ala8]-NKA(4-10) and neuropeptide gamma (NP gamma) all stimulate secretory responses in rat descending colon mucosa under voltage clamp conditions. Secretory responses (measured as short circuit current under voltage clamp conditions) were transient and those evoked by SP, SMSP, NKA and senktide were significantly reduced by pretreating tissues with the chloride channel blocker, diphenylamine carboxylate (DPC). Concentration-response curves showed varying degrees of sensitivity to tetrodotoxin (TTX). Senktide-induced secretion was virtually abolished by TTX, while NP gamma and [beta Ala8]-NKA(4-10) were not significantly altered. Rightward shifts of concentration-response curves were observed for SMSP, NKA and SP in TTX treated preparations compared with controls. NKA response curves in the presence of TTX were further inhibited by MEN10,207 and CP-96,345. GR71251, GR82334 and CP-96,345 all inhibited SMSP secretory responses with pA2 values of 5.8, 6.5 and 6.9 respectively. In conclusion three types of neurokinin receptor exist in preparations of rat colon mucosa and their relative location within neuronal and epithelial surfaces are discussed.

Lack of evidence of omeprazole genotoxicity in Sprague-Dawley rats.

Omeprazole is a proton pump inhibitor of increasingly wide use in the treatment of peptic ulcers. Although omeprazole has been subjected to an extensive range of genotoxicity tests, which have all been concluded as negative, the ability of this compound to interact with DNA and elicit unscheduled DNA synthesis in the rat gastric mucosa has been the subject of debate. Therefore, we have examined omeprazole using other genotoxicity end-points. In female Sprague-Dawley rats, the administration by the oral route of 100 mg/kg, either as neutral (pH 7.0) suspension or as suspension acidified to pH 1.5, which favours its transformation into the active form of sulphenamide, did not induce DNA fragmentation in gastric mucosa and liver, as detected by the alkaline elution technique. In the same experimental conditions, a frequency of total nuclear anomalies (micronuclei, pyknosis and karyorrhexis) that was significantly higher than in controls was detected with both types of suspension in forestomach and descending colon mucosa. However, in both tissues this higher frequency of nuclear anomalies was mostly due to pyknosis and karyorrhexis, which may be the outcome of a non-genotoxic effect, whereas there was no significant increase in the number of micronucleated cells, and this suggests the absence of clastogenic activity. Finally, in rats initiated with N-nitrosodiethylamine, the oral administration of 100 mg/kg omeprazole for 14 successive days produced a modest but statistically significant increase of liver gamma-glutamyltranspeptidase positive foci, which is consistent with a potential promoting activity.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of capsaicin upon electrogenic ion transport in rat descending colon

Preparations of rat descending colon mucosa have been used to record changes in short circuit current (SCC) under voltage clamp conditions. When added to the basolateral compartment capsaicin (8-methyl-N-vanillyl-6-nonenamide, 0.1-3 microM) caused an initial transient increase in SCC, followed by a more prolonged reduction in SCC, that lasted for 20-30 min. Repeated applications of 3 microM capsaicin caused desensitisation of the initial secretory response. The antisecretory effects (i.e. reduction in SCC from the original baseline) remained, although they were significantly reduced. In some preparations described as "non-responders", 3 microM capsaicin did not elicit a secretory response. No desensitization of the remaining antisecretory responses was observed in these tissues; in fact these reductions in SCC were consistently larger than those from tissues which responded with a secretory response. Tetrodotoxin (100 nM), hexamethonium (10 microM), and yohimbine (50 microM) had no significant effect upon either secretory or antisecretory responses. Ruthenium red (10 microM) abolished the secretory response to 3 microM capsaicin, but had no effect upon the antisecretory responses. Pretreatment of the tissues with 1 microM substance P (SP) resulted in significant desensitisation to the peptide and abolished the secretory response to 3 microM capsaicin. The antisecretory responses remained, and were significantly larger compared with responses from control tissues.

The antisecretory effects of somatostatin and analogues in rat descending colon mucosa

Somatostatin-14 (SS-14) and somatostatin-28 (SS-28) produce concentration dependent reductions in short-circuit current in rat colonic mucosa. EC50 values of 15.0 and 13.3 nM were obtained for SS-14 and SS-28 respectively while the N-terminal fragments of SS-28, namely somatostatin-(1-12) (SS1-12) and somatostatin-(1-14) (SS1-14) were inactive. Cyclo(Pro-Phe-D-Trp-Lys-Phe) and cyclo(Pro-Tyr-D-Lys-Thr-Phe) were potent antisecretory peptides, like SS-14 and SS-28; while the putative somatostatin antagonist, cyclo(7-aminoheptanoyl-Phe-D-Trp-Lys-Thr[Bzl]) exhibited neither agonist nor antagonist effects. Responses to SS-14 could be regulated by agents which affected the secretory state of the epithelium. Antisecretory effects of SS-14 were markedly attenuated by piroxicam and were restored following piroxicam plus either forskolin or vasoactive intestinal polypeptide (VIP). SS-14 also attenuated secretory responses produced by carbachol, substance P (SP), VIP and α- and β-calcitonin gene related peptide (α-, β-CGRP). Therefore, SS-14 exhibits broad spectrum antisecretory effects in rat descending colon mucosa.

Effects of Continuous Administration of 1,25-Dihydroxyvitamin D3 on Plasma Minerals and Unoccupied Colon Mucosal 1,25-Dihydroxyvitamin D3 Receptor Concentrations

Six mature nonlactating, nonpregnant Jersey cows were implanted with Alzet mini-osmotic pumps, which delivered 50 μg of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] each day for 7 d in an effort to mimic plasma concentrations of 1,25-dihydroxyvitamin D [1,25-(OH)2D] observed in cows at parturition. Plasma samples were obtained daily beginning 6 d prior to implantation and ending 8 d after removal of the implants. Six biopsies of the descending colon mucosa were obtained per rectum before and after implantation and assayed for unoccupied 1,25-(OH)2D3 receptor concentration. Plasma concentration of 1,25-(OH)2D increased from 37 pg/ml pretreatment to 294 pg/ml with the pumps implanted. Plasma Ca concentration increased within 2 d after implantation and remained elevated for 7 d after the pumps were removed. Unoccupied colon mucosa 1,25-(OH)2D3 receptor mean concentration prior to treatment was 14.6 fmol/mg protein and increased within 2 d following implantation to 30.5 fmol/mg protein. These data suggest that 1,25-(OH)2D3 upregulates its own receptor in the intestine of the cow.