Zarko Rasic

Stable gastric pentadecapeptide BPC 157 heals rectovaginal fistula in rats

AIM Rectovaginal fistula is a devastating condition providing more than 99% of patients for surgical treatment. We hypothesized that rectovaginal fistula may be healed by therapy with stable gastric pentadecapeptide BPC 157, in consistence with its initial clinical application and effect on external fistulas. MAIN METHODS BPC 157 (10μg/kg or 10ng/kg) was given perorally, in drinking water (0.16μg/ml or 0.16ng/ml, 12ml/rat/day) till sacrifice, or alternatively, intraperitoneally, first application at 30min after surgery, last at 24h before sacrifice. Controls simultaneously received an equivolume of saline (5.0ml/kg ip) or water only (12ml/rat/day). The assessment (i.e., rectal and vaginal defect, fistula leakage, defecation through the fistula, adhesions and intestinal obstruction as healing processes) was at day 1, 3, 5, 7, 10, 14 and 21. KEY FINDINGS Regularly, rectovaginal fistulas exhibited poor healing, with both of the defects persisting, continuous fistula leakage, defecation through the fistula, advanced adhesion formation and intestinal obstruction. By contrast, BPC 157 given perorally or intraperitoneally, in μg- and ng-regimens rapidly improved the whole presentation, with both rectal and vaginal defects simultaneously ameliorated and eventually healed. The maximal instilled volume was continuously raised till the values of healthy rats were achieved, there were no signs of defecation through the fistula. A counteraction of advanced adhesion formation and intestinal obstruction was achieved. Microscopic improvement was along with macroscopic findings. SIGNIFICANCE BPC 157 effects appear to be suited to induce a full healing of rectovaginal fistulas in rats.

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.

W1337 Therapy With Gastric Pentadecapeptide BPC 157 (PL14736) and L-NAME in Short Bowel Syndrome and Entero-Enteral Anastomosis Healing in Rats

Gastric pentadecapeptide BPC 157 (BPC 157), safe in trials for inflammatory bowel disease (IBD) (PL14736 Pliva)(Gastroenterology, 2005), could also be useful in IBD complications. It successfully heals the intestinal anastomosis in rats (Surg Today, 2007) and has a therapeutic effect on the short-bowel syndrome that compromises nutritional status and healing process (Dig Dis Sci, 2008). Also, it interacts with the NO-system and counteracts the effects of NOS-inhibition, induced by application of NOS-inhibitor, N-G-nitro-L-arginine-methyl ester (L-NAME) (Eur J Pharmacol, 1997, J Pharm Sci, 2008). Thereby, this study investigates whether the therapeutic effect of the pentadecapeptide BPC 157 on the short-bowel syndrome could be abolished by L-NAME application. Extensive small bowel resection (from the 4th ileal artery cranially of the ileocecal valve to 5 cm underneath the pylorus) and entero-enteral anastomosis were carried out in male rats as described before (Surg Today, 2007, Gastroenterology, 2007) Medication (BPC 157 (10 µg, 10 ng, 10 pg), L-NAME (5 mg) or an equivolume of saline (5 ml) was given once daily, i.p. (/kg), first application 30 min following surgery, last 24 h before sacrifice (at the post-operative day 1, 3, 5, 6, 7, 14). The assessment includes macroscopic presentation (body weight, mass of small intestine (3 cm cranially to 3 cm caudally from anastomosis), diameter measurements (jejunal, ileal and anastomosis diameter), biomechanical presentation (the volume (ml) instilled through a syringe-perfusion pump system (1ml /10 sec)) to the induction of leakage), microscopic presentation (Surg Today, 2007, Dig Dis Sci, 2008). A severe bowel syndrome was observed (severe body weight loss, jejunal diameter increase, a small volume (2-3 ml) before leakage, particularly decreased height of the villi distal to anastomosis). This was completely counteracted by both pentadecapeptide BPC 157 regimens. Weight gain to healthy values along with markedly higher volume values (4-6 ml) was obtained without leakage. Anastomosis healing is improved (edema markedly attenuated, granulocyte number decreased, necrosis attenuated, granulation tissue, reticulin and collagen formation markedly increased, and finally epithelization increased). All measured diameters were similar to diameter of healthy animals, the height of the villi distal to the anastomosis were higher than in controls. BPC 157 therapeutic effect could be not abolished by L-NAME application. Alone, L-NAME application aggravated short bowel syndrome. Short bowel syndrome and entero-enteral anastomosis healing may be a particular target for therapy with BPC 157.