Nermin Lojo

Effects of Diclofenac, L-NAME, L-Arginine, and Pentadecapeptide BPC 157 on Gastrointestinal, Liver, and Brain Lesions, Failed Anastomosis, and Intestinal Adaptation Deterioration in 24 Hour-Short-Bowel Rats

Stable gastric pentadecapeptide BPC 157 was previously used to ameliorate wound healing following major surgery and counteract diclofenac toxicity. To resolve the increasing early risks following major massive small bowel resectioning surgery, diclofenac combined with nitric oxide (NO) system blockade was used, suggesting therapy with BPC 157 and the nitric oxide synthase (NOS substrate) L-arginine, is efficacious. Immediately after anastomosis creation, short-bowel rats were untreated or administered intraperitoneal diclofenac (12 mg/kg), BPC 157 (10 μg/kg or 10 ng/kg), L-NG-nitroarginine methyl ester (L-NAME, 5 mg/kg), L-arginine (100 mg/kg) alone or combined, and assessed 24 h later. Short-bowel rats exhibited poor anastomosis healing, failed intestine adaptation, and gastrointestinal, liver, and brain lesions, which worsened with diclofenac. This was gradually ameliorated by immediate therapy with BPC 157 and L-arginine. Contrastingly, NOS-blocker L-NAME induced further aggravation and lesions gradually worsened. Specifically, rats with surgery alone exhibited mild stomach/duodenum lesions, considerable liver lesions, and severe cerebral/hippocampal lesions while those also administered diclofenac showed widespread severe lesions in the gastrointestinal tract, liver, cerebellar nuclear/Purkinje cells, and cerebrum/hippocampus. Rats subjected to surgery, diclofenac, and L-NAME exhibited the mentioned lesions, worsening anastomosis, and macro/microscopical necrosis. Thus, rats subjected to surgery alone showed evidence of deterioration. Furtheremore, rats subjected to surgery and administered diclofenac showed worse symptoms, than the rats subjected to surgery alone did. Rats subjected to surgery combined with diclofenac and L-NAME showed the worst deterioration. Rats subjected to surgery exhibited habitual adaptation of the remaining small intestine, which was markedly reversed in rats subjected to surgery and diclofenac, and those with surgery, diclofenac, and L-NAME. BPC 157 completely ameliorated symptoms in massive intestinal resection-, massive intestinal resection plus diclofenac-, and massive intestinal resection plus diclofenac plus L-NAME-treated short bowel rats that presented with cyclooxygenase (COX)-NO-system inhibition. L-arginine ameliorated only L-NAME-induced aggravation of symptoms in rats subjected to massive intestinal resection and administered diclofenac plus L-NAME.

Stable gastric pentadecapeptide BPC 157 in the treatment of colitis and ischemia and reperfusion in rats: New insights

AIM To provide new insights in treatment of colitis and ischemia and reperfusion in rats using stable gastric pentadecapeptide BPC 157. METHODS Medication [BPC 157, L-NAME, L-arginine (alone/combined), saline] was bath at the blood deprived colon segment. During reperfusion, medication was BPC 157 or saline. We recorded (USB microscope camera) vessel presentation through next 15 min of ischemic colitis (IC-rats) or reperfusion (removed ligations) (IC + RL-rats); oxidative stress as MDA (increased (IC- and IC + RL-rats)) and NO levels (decreased (IC-rats); increased (IC + RL-rats)) in colon tissue. IC + OB-rats [IC-rats had additional colon obstruction (OB)] for 3 d (IC + OB-rats), then received BPC 157 bath. RESULTS Commonly, in colon segment (25 mm, 2 ligations on left colic artery and vein, 3 arcade vessels within ligated segment), in IC-, IC + RL-, IC + OB-rats, BPC 157 (10 μg/kg) bath (1 mL/rat) increased vessel presentation, inside/outside arcade interconnections quickly reappeared, mucosal folds were preserved and the pale areas were small and markedly reduced. BPC 157 counteracted worsening effects induced by L-NAME (5 mg) and L-arginine (100 mg). MDA- and NO-levels were normal in BPC 157 treated IC-rats and IC + RL-rats. In addition, on day 10, BPC 157-treated IC + OB-rats presented almost completely spared mucosa with very small pale areas and no gross mucosal defects; the treated colon segment was of normal diameter, and only small adhesions were present. CONCLUSION BPC 157 is a fundamental treatment that quickly restores blood supply to the ischemically injured area and rapidly activates collaterals. This effect involves the NO system.

The Effect of Pentadecapeptide BPC 157 and High Dose Diclofenac on Induced Short Bowel Syndrome

Stable gastric pentadecapeptide BPC 157 (an anti-ulcer peptide effective in IBD trials, LD1 not achieved) counteracted short bowel syndrome (Sever et al., Dig Dis Sci, 2009) and protected rats against diclofenac gastrointestinal injuries (Ilic et al., J Physiol Pharmacol, 2009). Aim: In this study we tested the effect of pentadecapeptide 157 on induced short bowel syndrome rats aggravated with high dose of diclofenac. Material and methods We used 80%-small intestine resection and then short bowel syndrome rats received diclofenac application (12.5 mg/kg i.p.). Medication (BPC 157 10 μg/kg or an equivolume of saline (5 ml/kg ip) was given immediately after diclofenac. Animals were sacrificed 24 h after the surgery. Assessment included bleeding period (sec), gastrointestinal lesions (sum of longest diameters of lesions stomach, small and large intestine), serum AST, ALT, bilirubine levels. Microscopic analysis was also performed. Results All BPC 157 treated animals had shorter bleeding period compared to control group animals (210 +/- 37s vrs. 480 +/- 43 s). Bilirubine levels were significantly increased in control group animals compared to BPC 157 treated animals ( 192.3 +/- 46.7 μmol/ L vrs 6.8+/- 3.1 μmol/L). AST, ALT, LDH were increased in both tested groups (AST: 308+/-14 u/L (BPC157) vrs 383+/-27 u/L (con.) ; ALT: 52+/-4 u/L (BPC157) vrs 63+/-6 u/L (con.)) Number of gastrointestinal lesions (gastric, duodenal, small and large intestinal ulcerations) was significantly larger in control animal group compared to BPC 157 treated animals (gastric: 5.6 +/- 2.7 lesions (con) vrs . 0.5+/-0.2 lesions (BPC) ; duodenal (1.3 +/-0.2 lesions(con) vrs 0 lesions (BPC) ; jejunum, ileum and rectum score (1- ulcerations, 2- erosions and erithema, 3- normal mucosal surface) ; Jejunum: 1.6+/- 0, 7(con) vrs 2.8+/-0.2 (BPC) ; Ileum: 1 +/- 0, 4 (con) vrs 2.8+/-2 (BPC) ; Rectum 1+/-0.3 (con) vrs 2.6 +/- 0, 4 (BPC)). Conclusion According to our results we could conclude that BPC 157 improves small bowel and liver lesions healing in malnutrition conditions even during high dose diclofenac exposure.

Su1279 The Beneficial Effect of Pentadecapeptide BPC 157 on Healing of Rectovaginal Fistulas in Rats

G A A b st ra ct s who tend to be non-ambulatory, older, and more frail, intolerance and thus, noncompliance with the bowel purge, is a critical factor influencing the quality of bowel prep during colonoscopy. Aims: To evaluate patient compliance, feasibility and efficacy of a split-dose PEG-based lavage in the inpatient setting. Methods: Single-center, prospective observational pilot study of a consecutive convenience sample comparing inpatients that received 4-L PEG solution versus split dose solution (2-L PEG the evening prior and 2-L after midnight on the day of the procedure). The Boston Bowel Preparation Scale was used to evaluate the quality of the bowel prep. Tolerability, patient preferences and adverse events were recorded using a self-administered patient survey completed prior to colonoscopy and through chart review. The Students t test was used for continuous variables and the Fishers exact test was used for all categorical variables. A p value less than 0.05 was considered significant. Results: Patient demographics are shown in Table 1. All patients in the split dose group (SPLIT) completed the bowel prep successfully while 4 patients in the full dose group (FULL) did not; resulting in cancellation of the procedure. The majority of patients in the SPLIT group (85%) would prefer split dose over full dose for future colonoscopies while 39% of patients in the FULL group would not choose full dose lavage again. There were no significant differences in adverse events or quality of bowel prep between the two groups (Table 2). Conclusions: This pilot study demonstrates that the split dose 4-L PEG solution is a well-tolerated, feasible bowel cleansing method among hospitalized patients. The higher rates of completion of the bowel prep among the split dose bowel cleansing group suggest that split dose lavage may enhance compliance with bowel prep before colonoscopy among hospitalized patients. Table 1. Baseline Characteristics of Patients

Mo2065 The Effect of Pentadecapeptide BPC 157 on Duodenocolic Fistula Healing

Background: Keratins (K) are intermediate filaments important for cell protection from stress and for protein targeting in intracellular pathways. Keratins in epithelia forming the intestinal wall consist of type II (K7, K8) and type I (K19, K18, K20) heteropolymers, where K8 and K19 are the main components. In humans, keratin mutations predispose to liver disorders, while the roles of keratins in intestinal diseases are still ambiguous. Support for keratins in intestinal health comes from the K8-null (KO) mouse manifesting early chronic colitis, similar to the human inflammatory bowel disease a major risk factor for developing colorectal cancer (CRC).We hypothesize that the K8-null related colitis would predispose mice to CRC. Further support for keratins in the CRC progression are that K8KO colonocytes hyperproliferate, are resistant to apoptosis, have abnormal differentiation patterns and interact with mitogen activated kinases. Aims: The aim was to test whether aging K8KO mice with chronic colitis have spontaneous neoplastic changes, and if the K8KO induced colonic changes render mice more susceptible to induced CRC than K8 wild type (WT). Methods and Results: 7-9 month old K8KO and WT untreated mice displayed no colon adenomas, and rare abnormal crypt foci as studied by methylene blue staining and hematoxylin and eosin staining. A low dose of the colon carcinogen azoxymethane (AOM; 10 mg/kg, 1 injection, 3 months) induced aberrant crypt foci in both strains with a trend of higher incidence in K8KO distal colon compared to WT. A high dose of AOM (15 mg/kg, 4 weekly injections) induced after 10 weeks multiple distal colon tumors (1.5-4 mm in diameter) in both genotypes, with a dramatically higher number of tumors in the K8KO distal colon (20 ± 7.5 tumors per mouse) compared to WT (4 ± 2.9 tumors per mouse). Histological analysis showed that K8KO tumors were tubular adenomas of moderate grade while the slightly smaller WT tumors where low grade dysplasias. The dysplastic changes in all tumors showed translocation of beta-catenin from the cell membrane to nuclei indicating increased Wntsignaling. Interestingly, during basal non-tumor conditions, all K8KO colonic crypt epithelial cells stained strongly for membraneous beta-catenin, while in WT preferentially brushborder epithelial cells were positive. Conclusions: K8-null mice with chronic colitis and multiple hallmarks of cancer, are not predisposed to spontaneous dysplasia or CRC but have a highly increased susceptibility to carcinogen-induced adenomas. These results show that epithelial keratin-deficiency-induced colonic changes are risk factors in colorectal tumorigenesis.