Extra virgin olive oil-enriched diets protects the NSAID-induced gastric damage in rats: Role of leukocyte adherence

Ap il 1998 o ampli y a 820 base-pai egion o he u eC gene. The PCR p oduc s we e diges ed wi h he es ic ion endonucleases Sau3A and CJbl, and he agmen s gene a ed we e analyzed by aga ose gel elec opho esis. P esence o mul iple s ains o H pylo i was de ined when he sum o he es ic ion agmen s exceeded 820 bp. Resul s: H pylo i could be isola ed om 28 pa ien s (20 om gas ic biopsy and 8 om gas ic aspi a e samples); PCR on H. pylo i genomic DNA was posi i e in all o hem. When PCR was done di ec ly om gas ic biopsy/aspi a e samples, 24 (18 om gas ic biopsy and 6 om gas ic aspi a e samples) o hese pa ien s we e posi i e. No alse-posi i e esul was no ed. Fi e RFLP pa e ns wi h Sau3A and 3 RFLP pa e ns wi h CJbl we e iden i ied. RFLP pa e ns sugges ing p esence o mul iple s ains we e no ed in 3 pa ien s, when PCR was done on genomic DNA om Hpylo i isola es. PCR-RFLP pa e ns di ec ly om gas ic biopsies and aspi a es also iden i ied hese 3 pa ien s as ha bo ing mul iple s ains, and was indica i e o single s ains in he es 21 pa ien s. Conclusions: These esul s indica e ha PCR ampli ying he 820-bp egion o u eC di ec ly om gas ic biopsy and gas ic aspi a e samples is highly speci ic (100%) compa ed o ha om H pylo i genomic DNA; howe e he sensi i i y is 86%. PCR-RFLP analysis om H pylo i genomic DNA and di ec ly om gas ic biopsy and gas ic aspi a e samples is equally sensi i e in de ec ing simul aneous gas ic coloniza ion by mul iple s ains o H pylo i. • G0269 EXTRA VIRGIN OLIVE OIL-ENRICHED DIETS PROTECTS THE NSAID-INDUCED GASTRIC DAMAGE IN RATS: ROLE OF LEUKO- CYTE ADHERENCE. Ba anco M.D., Ala c6n de la Las a C., M0 il a V,, Ma n M.J., *Ga c a-Mau i io S., *S inchez-Ma gale V., *Es eban J., *He edas J.M. Dep . de Fa macolog a, Facul ad de Fa macia y Hospi al Uni e si a io Vi gen Maca ena, Uni e sidad de Se illa, Se illa, Spain. BACKGROUND. The Medi e anean die , which is cha ac e ized by a high in ake o an ioxidan s, ce eals and oli e oil, is epu ed o ha e an i- in lamma o y p ope ies. Oli e oil con ains a small amoun n-6 polyunsa u a ed a y acid bu he highes concen a ion o oleic acid, a monounsa u a ed a y acid, o all edible oils. Polyphenolic compounds a e also p esen in he ex a i gin oli e oil (un e ined oli e oil om oli es o good quali y) and he e is an in e es because hei an ioxidan ac i i ies. P e ious s udies o possible mechanisms o phenol ac ion indica e ha hese compounds a e able o sca enge ee adical and o b eak pe oxida i e chain eac ion. In addi ion, polyphenols exe se e al indi ec e ec s educing he p oduc ion o chemo ac ic and in lamma o y compounds. Ulce a ion in he gas oin es inal ac induced by NSAID is he majo limi a ion o hei he apeu ical use. A ascula e iology has been p oposed wi h ac i a ion o polymo phonuclea leukocy es. Neu ophil ac i a ion also induces changes in he epe oi e o cell su ace adhesion ecep o s and exp ession o he in eg ins a e in ol ed in neu ophil ex a asa ion du ing in lamma ion. Ma gina ion o ci cula ing PMN in o he gas ic mic oci cula ion is an ea ly and c i ical e en in he pa hogenesis o NSAID. AIMS. To examine he hypo hesis ha die s supplemen ed wi h ex a i gin oli e oil may educe he se e i y o he NSAID induced gas ic lesion and o explo e he e ec o some oli e oil polyphenols on quan i a i e and quali a i e changes in leukocy e adhesion ecep o s. METHODS. Weanling a s we e main ained on semisyn he ic die s o 6 weeks; s anda d die con aining 5% (w/w) o a as con ol and oli e oil suplemmen ed die s (5% and 20% w/w). Gas ic lesion was induced on he las day by o al adminis a ion o indome hacin (IND 60 mg/Kg b.w.). The leukocye in il a ion in gas ic wall was measu ing by he myelope oxidase ac i i y (MPO). The exp ession o in eg ins du ing neu ophil ac i a ion wi h FMLP was assessed by low cy ome y and he ollowing Mab we e used: TPI/40 an i-CDlla, Beam an i-CDllb, HCI/1 an i CDllc, and KIMI27 an i-CD18. The polyphenolic compounds assayed (25 laM - 1 mM) we e: oleu opein (OLR) and ca eic (CAF), sy ingic (SYR) and p o oca echuic (PRT) acids. RESULTS. In animals consuming s anda d die , he o al a ea o lesions was 14.7 ± 3.4 mm 2. In con as , in animals ed oli e oil die s gas ic damage dec eased in magni ude in pa allel wi h he die a y con en in he a . The ulce index was dec eased o 7.7 +_ 1.9 mm 2 (p < 0.01 s IND s anda d die ) eeding o 5% oli e oil en iched die and o 2.7-+ 0.8 mm 2 in animals consuming 20% oli e oil die (p <0.001 s IND s anda d die ). These p o ec i e e ec we e speci ically ela ed o a educ ion o neu ophil in il a ion (MPO alues). CAF, SYR and PRT induced a d ama ic dec ease o CD1 lb and CD1 lc exp ession (p < 0.001), whe eas a mode a e dec ease was obse ed wi h OLR (CD 1 lc, p < 0.05). In con as , he exp ession o o he adhesion molecules was una ec ed (CD1 la, CD18). CONCLUSION. Resul s demons a e he p e en i e p ope ies o ex a i gin oli e oil die s in NSAID induced gas ic mucosal inju y. This e ec could be explained by i s in i o an in lamma o y p ope ies bu also by he educ ion o he in i o exp ession o cell adhesion molecules. Esophageal, Gas ic, and Duodenal Diso de s A67 G0270 H. PYLORI INFECTED MUCOSA IN GASTRIC ULCER SAMPLES: PHOSPHOLIPASE ALCOHOL DEHYDROGENASE AND UREASE ACTIVITIES. R. Ba e o-Zu iiga 1,3, M. Okuyama 2, Y. Ka o 3, F. Ma o a 3,6, H. Oh a 4, T. TakekoshP, M. Ma uyama 3, D. Mu guia I. GI Se ice Mexico Gene al Hosp. 1, Ins i . o Whole Body Me abolism, Chiba, Japan 2, GI Se ice, S. Anna Hosp., Como, I aly. 6, In . Med. 3, Su ge y 4 and Pa hology 5 Dep . Cance Ins i u e Hosp. Tokyo, Japan. Obee i e: A mucosal su ace H. pylo i (HP) enzymes gene a e oxic molecules: ammonia (u ease, UR), lysoleci hin (phospholipase, PL) and ace aldehyde (alcohol dehyd ogenase, ADH). We in es iga ed whe he UR, PL and ADH ac i i ies a e al e a ed in he gas ic mucosa om gas ic ulce (GU), compa ed wi h con ols. Me hods: Biopsy aken om 44 GU and 73 con ols, comp ising wo subg oups: 48 non ulce pa ien s (NUG) and 24 pa ial gas ec omy pa ien s (PG) as gas ic inju ed con ol. The HP s a us we e de ec ed by cul u e, in i o u ease and his ological es s. Enzyme ac i i ies we e de ec ed by newly adio ace echnique TLC- Au o adioluminog aphy (TLC-ARLG) Resul s: The mean o enzymes le els in HP posi i e samples shows s a is ical signi ican ly di e ences, han HP nega i e. [*Pi: S uden 's es o pai ed da a; P ob. (2- ail)]. H. pylo i (+) H. pylo i (-) Mean ± SE Mean ± SE Pi PL .116 _+ .22 .179 + .38 .09* ADH .138 ± .106 .195 ± .12 .01" U ease 5.79 +_ 3.73 .598 ± .676 .004* The mos e iden al e a ions o PL we e induced by PG and UG (84% and 34% less han NUG espec i ely). The GU samples in ec ed wi h HP had signi ican ly lowe ADH (mean GU=0.151 s. NGU=0.285 pCi/mg/min) and PL ac i i ies [(mean GU=0.116 s. NGU=0.179 (min-lX100)] han NGU samples. The mean u ease le els in HP posi i e samples we e signi ican ly highe han HP nega i e samples (Table). Al e a ion o enzyme ac i i ies we e well co ela ed wi h he deg ee o mucosal changes such as mononuclea o polymo phonuclea cell in il a ion. Conclusion: The UR, PL and ADH enzyma ic p o iles, e lec he pa hological adap a ions behind mucosal inju y in UP and PG. Al hough high ac i i y o UR indica es well he p esence o HP, impai men ac i i ies o PL and ADH e lec mo e he gas ic mucosal in lamma ion han HP in ec ion "pe se". Fu he s udies should be p ima ily a emp ed wi h TLC- ARLG in pa ien s wi h gas ic disease and HP ea men . This esea ch was unded by The Founda ion o Li e Science Resea ch, Japan • G0271 INDOMETHACIN (Indo) AND BILE SALTS (BS) COMPETE FOR THE BILIARY PHOSPHATIDYCHOLINE (PC): AN EXPLANATION OF Indo-INDUCED INTESTINAL INJURY. JM Ba ios ° and LM Lich enbe ge *, °Depa men o Pedia ics, Baylo College, Hous on TX and *Depa men o In eg a i e Biology, Uni e si y o Texas Medical School, Hous on TX. Backg ound: PC ep esen s ± 40% o he o ganic ma e ial o bile, PC has he capaci y o associa ing wi h non-s e oidal an i-in lamma o y d ugs (NSAIDs) and bile sal s, dec easing he GI oxici y o hese wo classes o compounds. NSAIDs ha unde go en e ohepa ic cycling a e oxic o he ileal mucosa, by a mechanism ye o be elucida ed. Hypo hesis: Bilia y PC associa es wi h and de oxi ies bile sal s, o ming mixed micelles. NSAIDs sec e ed in o he bile compe e wi h bile sal s o he a ailable PC, esul ing in inc ease in he concen a ion o ee bile sal o damage he in es inal mucosa. Me hods: 5 aM o Deoxycholic acid, Indo and PC, we e ins illed in o a loop o he dis al ileum o anes he ized a s, alone and in combina ion. A e 30 minu es, loop luid and ileal mucosa we e collec ed o hemoglobin (Hb) and con ac angle analysis. The same combina ions we e used o assess hei e ec on human e y h ocy es (RBCs) as measu ed by deg ee o hemolysis, excep PC was adminis e ed a bo h 5 and 10 aM. Resul s: a e shown below wi h *=p < 0.05 s saline/bu e . ] ] Saline ] BS ] PC ] Ileal Hyd ophobici y (Con ac 0) i Sal oe I I I I Indo 23.5 -+ 3.2 20.3 -+ 3.4 14.1 -+ 2.6* Ileal Loop Hemoglobin Concen a ion ( ag %) [Saline [5.5- 1.6152.7±18.8" 5.7±1.0[ Indn 3.0 + 1.0 9.8 =1:3.8 17.8 + 5.0* RBC Hemolysis I [ Saline [ BS BS + 5PC BS + 10PC Saline [ 0 [ 8.4 ± 1.5" 0.7 ± 0.1 0.7 +_ 0.3 Indo 0 8.8 +_ 1.3" 4.8 ± 2.6* 0.5 ± 0.6 In bo h, in i o and in in i o expe imen s he p o ec i e e ec s o PC agains bile sal -induced inju y we e e e sed by Indo. Conclusion: These indings con i m ou hypo hesis ha PC p o ec s agains he inju ious ac ion o bile sal s on cell memb anes. Indo and pe haps o he NSAIDs ha en e bile, damage he mucosa, no by a di ec oxic ac ion, bu by compe ing o he a ailable p o ec i e PC molecules.