. 2016 Nov 7;22(41):9127-9140.
doi: 10.3748/wjg.v22.i41.9127.
Ivka Djakovic 1 , Vedran Cesarec 1 , Goran Madzarac 1 , Tomislav Becejac 1 , Goran Zukanovic 1 , Domagoj Drmic 1 , Lovorka Batelja 1 , Anita Zenko Sever 1 , Danijela Kolenc 1 , Alen Pajtak 1 , Nikica Knez 1 , Mladen Japjec 1 , Kresimir Luetic 1 , Dinko Stancic-Rokotov 1 , Sven Seiwerth 1 , Predrag Sikiric 1
Affiliations
PMID: 27895400 PMCID: PMC5107594 DOI: 10.3748/wjg.v22.i41.9127
Esophagogastric anastomosis in rats: Improved healing by BPC 157 and L-arginine, aggravated by L-NAME
Zeljko Djakovic et al. World J Gastroenterol. 2016.
Abstract
Aim: To cure typically life-threatening esophagogastric anastomosis in rats, lacking anastomosis healing and sphincter function rescue, in particular.
Methods: Because we assume esophagogastric fistulas represent a particular NO-system disability, we attempt to identify the benefits of anti-ulcer stable gastric pentadecapeptide BPC 157, which was in trials for ulcerative colitis and currently for multiple sclerosis, in rats with esophagocutaneous fistulas. Previously, BPC 157 therapies have promoted the healing of intestinal anastomosis and fistulas, and esophagitis and gastric lesions, along with rescued sphincter function. Additionally, BPC 157 particularly interacts with the NO-system. In the 4 d after esophagogastric anastomosis creation, rats received medication (/kg intraperitoneally once daily: BPC 157 (10 μg, 10 ng), L-NAME (5 mg), or L-arginine (100 mg) alone and/or combined or BPC 157 (10 μg, 10 ng) in drinking water). For rats underwent esophagogastric anastomosis, daily assessment included progressive stomach damage (sum of the longest diameters, mm), esophagitis (scored 0-5), weak anastomosis (mL H2O before leak), low pressure in esophagus at anastomosis and in the pyloric sphincter (cm H2O), progressive weight loss (g) and mortality. Immediate effect assessed blood vessels disappearance (scored 0-5) at the stomach surface immediately after anastomosis creation.
Results: BPC 157 (all regimens) fully counteracted the perilous disease course from the very beginning (i.e., with the BPC 157 bath, blood vessels remained present at the gastric surface after anastomosis creation) and eliminated mortality. Additionally, BPC 157 treatment in combination with L-NAME nullified any effect of L-NAME that otherwise intensified the regular course. Consistently, with worsening (with L-NAME administration) and amelioration (with L-arginine), either L-arginine amelioration prevails (attenuated esophageal and gastric lesions) or they counteract each other (L-NAME + L-arginine); with the addition of BPC 157 (L-NAME + L-arginine + BPC 157), there was a marked beneficial effect. BPC 157 treatment for esophagogastric anastomosis, along with NOS-blocker L-NAME and/or NOS substrate L-arginine, demonstrated an innate NO-system disability (as observed with L-arginine effectiveness). BPC 157 distinctively affected corresponding events: worsening (obtained with L-NAME administration that was counteracted); or amelioration (L-arginine + BPC 157-rats correspond to BPC 157-rats).
Conclusion: Innate NO-system disability for esophagogastric anastomoses, including L-NAME-worsening, suggests that these effects could be corrected by L-arginine and almost completely eliminated by BPC 157 therapy.
Keywords: Aggravation; BPC 157; Curative treatment; Esophagogastric anastomosis; L-NAME; L-arginine; Rats.
PubMed Disclaimer
Conflict of interest statement
Conflict-of-interest statement: The authors state that they have no conflicts of interest.
Figures Figure 1
Gastric lesions, sum of the longest lesions diameters, mean ± SD, mm, in rats that underwent esophagogastric anastomosis. Medication (/kg) given intraperitoneally (ip) (once time daily) or continuously in drinking water (po) after the creation of an esophagogastric anastomosis in rats. BPC 157 (10 μg, 10 ng), L-NAME (5 mg), and L-arginine (100 mg) given alone and/or combined intraperitoneally with the first application at 30 min after anastomosis creation and the last at 24 h before sacrifice. Drinking water alone (12 mL/d per rat) or BPC 157 in drinking water (10 μg, 10 ng/kg; 0.16 μg, 0.16 ng/mL) was provided continuously until sacrifice. aP < 0.05, at least, vs control. Figure 2
Anastomosis strength. Water volume that could be sustained before leakage, mean ± SD, mL, in rats that underwent esophagogastric anastomosis. Medication (/kg) given intraperitoneally (ip) (once time daily) or continuously in drinking water (po) after the creation of esophagogastric anastomosis in rats. BPC 157 10 μg and 10 ng, L-NAME 5 mg, and L-arginine 100 mg given alone and/or combined intraperitoneally with the first application at 30 min after anastomosis creation and last at 24 h before sacrifice. Drinking water alone (12 mL/d per rat) or BPC 157 in drinking water (10 μg, 10 ng/kg; 0.16 μg, 0.16 ng/mL) was provided continuously until sacrifice. aP < 0.05, at least, vs control. Figure 3
Pressure in the esophagus at the anastomosis site. Mean ± SD, cmH2O, in rats that underwent esophagogastric anastomosis. Medication (/kg) given intraperitoneally (ip) (once time daily) or continuously in drinking water (po) after the creation of esophagogastric anastomosis in rats. BPC 157 10 μg and 10 ng, L-NAME 5 mg, and L-arginine 100 mg given alone and/or combined intraperitoneally with the first application at 30 min after anastomosis creation and last at 24 h before sacrifice. Drinking water alone (12 mL/d per rat) or BPC 157 in drinking water (10 μg, 10 ng/kg; 0.16 μg, 0.16 ng/mL) was provided continuously until sacrifice. aP < 0.05 at least vs control. The values of 68-76 cm H2O for the lower esophageal sphincter were considered to be normal, as determined previously[17,18,20-23]. Figure 4
Pressure in the pyloric sphincter. Mean ± SD, cmH2O, in rats that underwent esophagogastric anastomosis. Medication (/kg) given intraperitoneally (ip) (once time daily) or continuously in drinking water (po) after the creation of esophagogastric anastomosis in rats. BPC 157 10 μg and 10 ng, L-NAME 5 mg, and L-arginine 100 mg given alone and/or in combination intraperitoneally with the first application at 30 min after anastomosis creation and last at 24 h before sacrifice. Drinking water alone (12 mL/d per rat) or BPC 157 in drinking water (10 μg, 10 ng/kg; 0.16 μg, 0.16 ng/mL) was provided continuously until sacrifice. aP < 0.05, at least vs control. The values of 68-74 cm H2O for pyloric sphincter were considered normal, as previously determined[17,18,20-23]. Figure 5
Weight loss (g) presented as the Δ between the initial and final weights[13,18], mean ± SD, in rats that underwent esophagogastric anastomosis. Medication (/kg) given intraperitoneally (once time daily) or continuously in drinking water after the creation of an esophagogastric anastomosis in rats. BPC 157 10 μg and 10 ng, L-NAME 5 mg, and L-arginine 100 mg given alone and/or combined intraperitoneally with the first application at 30 min after anastomosis creation and the last at 24 h before sacrifice. Drinking water alone (12 mL/d per rat) or BPC 157 in drinking water (10 μg and 10 ng/kg; 0.16 μg and 0.16 ng/mL) was provided continuously until sacrifice. aP < 0.05, at least, vs control. Figure 6
Illustrative gross presentation of rats that underwent esophagogastric anastomosis at post-operative day 4 and then received medication saline (control, C), BPC 157 (B), or L-NAME (N). Figure 7
Illustrative microscopic presentation in rats that underwent esophagogastric anastomosis at post-operative day 2, hematoxylin eosin staining. Separated anastomosis edges (red arrows) with neutrophils within the edges (yellow arrow) were consistently noted in controls (c). Separated wound edges (red arrows). Connected edges in rats that underwent esophagogastric anastomosis and treated with BPC 157 regimens (black arrow) (b). Figure 8
Illustrative microscopic presentation in rats that underwent esophagogastric anastomosis at post-operative day 4 [controls (c), BPC 157 (b)] and at post-operative day 7 [BPC 157 (B)], hematoxylin eosin staining. Tissue destruction at the anastomosis edges (arrows) were consistently noted in controls (c), which is a presentation that precedes lethal outcome. Initial organization of the exudate (b) (arrows) and wound completely closed with granulation tissue (B) (arrows), which is consistently noted in rats that underwent esophagogastric anastomosis and were treated with BPC 157. Figure 9
Blood vessels presentation at the ventral site of the stomach surface in rats that just underwent esophagogastric anastomosis. In general, immediately after anastomosis creation and saline bath application, blood vessels disappeared from the rat gastric surface, and this effect lasts at least the next 15 min (scored Min/Med/Max 0/0/0), a period that was carefully monitored (control, c). With a BPC 157 bath immediately after anastomosis creation, the blood vessels did not disappear from the rat gastric surface; instead, these vessels remained present during at least the next 15 min of monitoring (BPC 157, b).
References
Sikiric P, Seiwerth S, Rucman R, Turkovic B, Rokotov DS, Brcic L, Sever M, Klicek R, Radic B, Drmic D, et al. Stable gastric pentadecapeptide BPC 157-NO-system relation. Curr Pharm Des. 2014;20:1126–1135. - PubMed
Seiwerth S, Brcic L, Vuletic LB, Kolenc D, Aralica G, Misic M, Zenko A, Drmic D, Rucman R, Sikiric P. BPC 157 and blood vessels. Curr Pharm Des. 2014;20:1121–1125. - PubMed
Sikiric P, Seiwerth S, Rucman R, Turkovic B, Rokotov DS, Brcic L, Sever M, Klicek R, Radic B, Drmic D, et al. Toxicity by NSAIDs. Counteraction by stable gastric pentadecapeptide BPC 157. Curr Pharm Des. 2013;19:76–83. - PubMed
Sikiric P, Seiwerth S, Rucman R, Turkovic B, Rokotov DS, Brcic L, Sever M, Klicek R, Radic B, Drmic D, et al. Focus on ulcerative colitis: stable gastric pentadecapeptide BPC 157. Curr Med Chem. 2012;19:126–132. - PubMed
Sikiric P, Seiwerth S, Rucman R, Turkovic B, Rokotov DS, Brcic L, Sever M, Klicek R, Radic B, Drmic D, et al. Stable gastric pentadecapeptide BPC 157: novel therapy in gastrointestinal tract. Curr Pharm Des. 2011;17:1612–1632. - PubMed