A 1 2 3 6 SSAT ABSTRACTS GASTROENTEROLOGY, Vol. 108 , No. 4

• MULTIPLE ORGAN CYTOKINE GENE EXPRESSION INDUCED BY ACUTE PANCREATITIS. J Norman. G Fink_ G Carter, A Rosemur~,v. M ~ . Dept of Surgery, University of South Florida, Tampa, FL.

Pro-inflammatory cytoklnes are elevated during acute pancreatitis and have been implicated in the progression of pauereatifis-assoeiated multiple organ dysfunction. Where these mediators are produced, however, is not known. This study was designed to examine pro-inflammatory eytokine gena expression in organs affected during panereatitis in order to investigate their role in propagating pancreatitis and multi-organ failure. Methods. Acute necrotizing pancreatitis was induced in adult male mice by IP injection of eaerulein (50 ~g/kg/hr x 4). Control mice received IP saline. Nine animals were sacrificed by exsanguination at 0, 0.5, 1, 2, 4, and 6 hours after the first caerulein injection. The pancreas, lungs, liver, kidneys, spleen, and gastroeaemius muscle were rapidly excised from all animals and prepared for mRNA and protein determination. The severity of pancreatitis was established by histologis grading and serum amylase and lipase. Serum IL-1, IL-6, and TNF-~ were determined by ELISA. The appearance of tissue cytoldnes was examined by Western blot analysis. Tissue cytokine mRNA was determined by rtPCR. Results. Caerulein injection was associated with histologie evidence of acute pancreatitis and elevation of serum amylase and lipase within the first hour (both p < 0.05 vs time 0; Student's t test), with the severity increasing through hour 6. There was no cytokine mRNA or protein detectable within serum, pancreas, liver, kidney, and muscle tissues prior to the induction of pancreatitis. TNF-~ mRNA was detected within the pancreas within one hour following the onset of panereatitis, with IL-1 and IL-6 mRNA appearing one and two hours later, respectively. The spleen demonstrated low constitutive expression of IL-1 and TNF-¢ n'LRNA which increased significantly by hour four (p<0,05). IL-1 and TNF-,~ mRNA became detectable within the lung and liver during the fourth hour ofpancreatitis with IL-6 mRNA following 2 hours later. Kidney and muscle failed to express cytokine mRNA at all times. Concomitant with mRNA prodnetion was the elevation of tissue levels of all three cytokines, detectable first within the pancreas, and then subsequently within the lung and liver (all p<0.05 vs Control). Similarly, IL-1, IL-6 and TNF-~ appeared in the serum beginning at hour 2 and increased through hour 6 (all p<0.005 vs Control). Conclusions. Pro-inflammatory cytokines are produced within the pancreas and within organs developing dysfunction during severe acute pancreatitis. The production of these cytokines appears to occur first within the pancreas and subsequently at distant sims. Serum IL-1, IL-6, and TNF-~z which correlate with pancreatitis severity are a result of systemic production at many sites.

THE INCREASED PRODUCTION OF INFLAMMATORY CYTOKINES BY DIFFERENT POPULATIONS OF ENTEROCYTES AFTER THERMAL INJURY. C.K. Oqle,PhD., J. Mao,MD, P-O Hasselqren,MD,PhD, J.W. Alexander,M.D.,Sc.D. University of Cincinnati, Department of Surgery and Shriners Burns Institute, Cincinnati, Ohio.

Purpose: To determine if distinct populations of normal and burn enterocytes respond differently to stimulation with LPS. Methods: Guinea pigs received a 30% flame burn and were sacrificed 24 hours after injury. Three populations of enterocytes were obtained by 3 extractions with EDTA (designated Fraction I (closest to the lumen), II, and III). The cells were incubated with LPS (10~g/ml) for 24 hours and supernatants were analyzed for IL-I and IL-6. Cell associated (ca)TNF was also determined since supernatant TNF values were negligible. Statistics: ANOVA (one between and two within) followed by Newman Keulls significance at p<0.05. Results (Values in u/ml):

Fraction I II III

Uni ts /ml -LPS + LPS -LPS + LPS -LPS + LPS TNF(ca) NO.15(a) O.30(a) 0.72 0.46(a) 1.04 2.80(a)

B 0.42 3.74 0.88 2.98 1.81 14.2(b)(c)

IL-1 N 232 268 184 289(a) 2 987(c}

B 318 596 441 688 471 1,371(b}(c) IL-6 N8.12 8.70(a} 6.15(a} 9.38 10.71(al 12.22(a)

B 10.7 16.1 12.7 13.2 14.1 19.7(b}(c)

Significant differences: (a) normal N vs burn B; (b) -LPS vs +LPS; (c) fraction I vs II vs IlI. Conclusions: Enterocytes furthest from the lumen (fraction III) produced more IL-I in responseto LPS before or after and more TNF and IL-6 after thermal injury. Thermal injury primed entero- cytes to produce more TNF, IL-I, and IL-6 in response to LPS. The increase in cell associated TNF after thermal injury could lead to increased cytotoxicity for enterocytes. The gut has the potential to become a potent source of inflammatory cytokines after thermal injury.

O SELECTIVE DO~GULATION OF GLUTAMINE TRANSPORT AFTER JEJUNOILEAL AUTOTRANSPLANTATION. A.J. Oishi*, Y. Inoue#, W.W. Scuba#, M.G. Sarr*. Dept. of Surgery, Mayo Clinic, Rochester, MN* and Massachusetts General Hospital, Boston, MAt.

Gut transplantation is complicated by weight loss and diarrhea. Little is known, however, about absorption of glutamine, an important substrate for enterocyte metabolism. Aim: To determine ileal transport of glutamine iF vitro and in vivo after intestinal transplantation. Methods: Brush-border membrane vesicles (BBMV) from the ileum of 6 dogs before and 2 and 8 wk after a model of jejunoileal autotransplantation were compared to 6 operated controls who had proximal and distal small bowel transection and reanastomosis to control for disruption of enteric neural continuity. Using a rapid mixing/ filtration technique, carrier-mediated transport of 100~M 3H-L-glutamine, glucose and other amino acids was assayed. In 6 other dogs, ileal absorp- tion of glutamine was determined in vive before and at 2 and 8 wk after this model using a triple lumen technique with isosmolar electrolyte solu- tion containing 20 mM glutamine. Results: Diarrhea occurred after this model of jejunoileal autotransplantation. BBMV transport of glutamine (but not glucose) decreased at 2 wk and remained decreased at 8 wk (113±6 vs 82±4 and 92±5 pmol/mg protein/10 sec, resp; each p<0.05) secondary to a decrease in V~x but not ~. Absorption of water and electrolytes decreased at 2 wk but returned to baseline at 8 wk. Glutamine absorption in vivo decreased at 2 and 8 wk (185±20 vs 121±27 and 151±22 ~mol/cm/min, resp; pc0.06; ANOVA). Conclu- sions: Jejunoileal autotransplantation down- regulated carrier-mediated glutamine transport and in vivo glutamine absorption (but not glucose) by decreasing the number of transporters and not by changing transporter affinity. This effect appears to be mediated by extrinsic denervation. Support: NIH DK39337 (MGS).

THIRTY-SIX YEARS' EXPERIENCE WITH ELECTIVE THERAPEUTIC PORTACAVAL SHUNT FOR BLEEDING ESOPHAGOGASTRIC VARICES DUE TO CIRRHOSIS. M,J. 0rloff. M.S. Orloff, S,L. Orloff. M. Rambotti B. Girard. Department of Surgery, UCSD Medical Center, San Diego CA.

Since 1958 we have performed elective therapeutic porta- caval shunt (PCS) (side-to-side in 92%, end-to-side in 8%) in 824 patients with biopsy-proven cirrhosis (alcoholic in 89%) who were usually referred to us from other hospitals after they recovered from bleeding esophagogastric varices (BEV). All patients have had at least 5 yr of follow-up. Age ranged from 27-79 yr, 67% were male, and 55% had had >i bleeding episode. Quantitative Child's risk classes were A-10%, B-58%, C-32%. BEV were proven in all patients by endoscopy and/or contrast radiography. Incidence of serious risk factors on admission or in past history was: ascites 62%; jaundice 55%; severe muscle wasting 40%; encephalopathy 33%; hyperdynamic state (CO~6L/min) 78%; past delirium tremens 15%. Mean PV-IVC pressure gradient (ram saline) pre-PCS was 251 and post-PCS was 22. Mean operative blood transfusions was 2.2u, and mean operative time was 3,2 hr (96%~4 hr). Survival to leave the hospital was 98.4% (operative mortality 1.6%). SuhseQuent survival rates were: i yr 95%, 5 yr 68%, i0 yr (actuarial) 62%, 15 yr (actuarial) 57%. Continued alcoholism caused 2/3 of the deaths. Encephalopathy occurred transiently in 10% and recurrently in 7%, compared to 33% pre-shunt. Long-term shunt patency was 99.6% by angiography and/or Doppler ultrasonography every 1-2 yr. Only 3 patients (0.4%) had recurrent varieeal bleeding, all due to shunt occlusion. 62% abstained from alcohol, and 89% of the abstainers survived 5 yr. LFT after 1 yr were improved in 72%, unchanged in 21%, and worse in only 7%. 67% resumed full- or part-time work. Conclusion: In the largest and longest experience reported to date, elective therapeutic PCS for BEV resulted in prolonged survival and life of satisfactory quality in 2/3 of cirrhotic patients. Liver transplanta- tion is being done with increasing frequency in bleeding alcoholic cirrhotics who practice abstinence, but PCS in abstainers results in substantially greater 5-yr survival (89%), with much lower morbidity and cost.