3035
induced Gas ic MuoosaJ Apop osis in Helioobac e Pylo i Posi i e Pa ien s Taking
Low Dose Aspi in: A B-Mon h Case-con ol S udy
Xinmin Zhou, K. Lai, S. Lam, Queen Ma y Hosp, Hung Kong; Hongbo Zhang, Xijing Hosp,
Xi'an People'S Rep O China; X. Huang, Xiaoming Fan, W. Hui, Queen Ma y Hosp, Hong
Kong; Daiming Fan, Xijing Hosp, Xi'an People'S Rep O China; Benjamin C. Y. Wong,
Queen Ma y Hosp, Hong Kong
Backg ound-Mos s udies showed ha
Helicobac e pylo i(HP)
in ec ion inc eased he a e o
gas ic mucosal p oli e a ion and apop osis. Non-s e oidal an Hn lamma o y d ugs (NSAIDs)
is also known o a ec epop osis o gas ic epi helial cells, bu he e is li le in o ma ion abou
in e ac ion be ween Hp in ec ion and NSAIDs adminis a ion. Aim-The aim o his s udy is o
in es iga e he dynamic balance be ween apop osis and p oli e a ion and gene ic changes in
subjec s s a ed on aspi in. Me hods-se ial biopsy specimens o he gas ic an um we e
ob ained du ing endoscopy om 12 Hp posi i e subjec s wi h e adica ion, 12 Hp posi i e
subjec s wi hou e adica ion and 12 Hp nega i e subjec s, all o whom we e s a ed on
100
mg o aspi in a ime O. Apop osis and p oli e a ion we e measu ed by TUNEL and PCI~
immunos aining. The exp ession o apop osis associa ed p o eins bel-2, bax, and CPP-32
was also de ec ed by immunohis ocbemical s aining. Resul s-The apop o ic index (AI)was
signi ican ly inc eased in Hp posi i e pa ien s wi hou e adica ion a e six mon h ( om
12.17±3.83 o 17.58±7.82, P<O.05); whe eas he e was no signi ican changes in Hp posi i e
pa ien s 6 mon hs a e e adica ion and in Hp nega i e pa ien s. Simila ly he e wa e signi ican
inc ease in p oli e a ion index (PI) and he a io o AI/PI only in Hp posiU e subjec s wi hou
e adica ion, accompanied by a ma ked inc ease in exp ession o apo~o ic p o ein bax. The
exp ession o bcl-2 and CPP32 emained unchanged in all g oups. Conclusions-These esul s
sugges ha ac i e Hp in ec ion and aspi in in ake inc ease apop osis and p oli e a ion o
gas ic epi helial cells, and upse he AI/PI balance. The addi i e e ec is blocked by ea men
o Hp. This may p o ide a molecula basis o e adica ion o Hp be o e ini ia ion o acpidn ea -
men .
3036
De elopmen o an Imp o ed Fo mula ion o PhosphaUdyicholine (PC) - Associa ed
Nons e oidal An i-in lamma o y
D ugs (NSAIDs)
Lena d M, Lich enbe ge , Jimmy J. Rome o, Uni o Texas Medical Sch, Hous on, 13(;
Sude shan K. Sanduja, Na u al The apeu ics Inc, Suga Land, TX
Backgound:
We ha e p e iously epo ed ha : he gas ic mucosa has hyd ophobic ha ie
p ope ies due o he p esence o PC and o he zwl e ionic pbospholipids wi hin and coa ing
he o e lying mucus gel laye ; and ha NSAIDs ha e an abili y o weaken his lining by
chemically associa ing wi h PC
(Na u e Mud
1:154,1995). Acco dingly. we ha e de eloped a
s a egy o p e en his in e ac ion,and as a consequence NSAID-induced opical inju y, by
p e-associa ing an NSAID wi h ei he pu i ied o syn he ic PC. Ea lie p e-clinical and clinical
s udies demons a ed ha aqueous suspensions o a numbe o PC-NSAIDs ha e bo h lowe
GI oxici y and enhanced he apeu ic ac i i y in compa ison o he unmodi ied d ugs
(Na u e
Mud; Am J Cas o
94:1818, 1999; JPET 277:2773,1996). In he p esen s udy we e alua ed
a new me hod o p epa e PC-NSAIDs ha has he ad an age ha he inal o mula ion is
bo h solid and cos -e ec i e.
Me hods:
PC-NSAIDs we e p epa ed by admixing a leci hin oil
con aining 35% PC wi h one o he ollowing NSAIDs: aspi in (ASA); ibop o en (IBU); o
indome hacin (INDO), un il a uni o m pas e was o med. These solid o mula ions we e hen
es ed in acu e oden models o : ASA-induced gas ic ulce a ion; IBU- and INDO - induced
GI bleeding; and
NSAID-induoed
analgesic
ac~ i y (RandalI-Selli o,A ch lo ~ 111:
409, 1957).
Resul s:
I was demons a ed ha gas ic ulce a ion was signi ican ly educed by
70.3±3.5% by PC-ASA, and GI bleeding was signi ican ly educed by 42.3±6.4% and
82.0±8.5% by PC-IBU and PC-INDO espec i ely, in compa ison o he alues o a s adminis-
e ed he unmodi ied d ugs.Consis en wi h hese obse a ions employing acu e models o
NSAID-induced inju y, we also de e mined ha PC-INDO induced less dia hea and signi ican ly
ewe in es inal lesions and adhesions han unmodi ied INDO du ing a 4 day ea neo pe iod.
The inc eased GI sa e y o he new PC-NSAID o mula ions was no a ibu able o a lowe
bioa allabili y o he NSAIDs when adminis e ed as a solid, as he analgesic ac i i y o PC-
ASA, PC-IBU and PC-INDO we e all signi ican ly enhanced in compa ison o he unmodi ied
d ugs.
Conclusion:
These esul s indica e ha he new cos -e ec i e me hod o o mula e
PC-NSAIDs p o ides a GI sa e, he apeu ically supe io p oduc o ea ch onic in lamma o y
diso de s. Long- e m clinical ials e alua ing he GI sa e y o his new amily o PC-NSAIDs
a e planned. (suppo ed by NIH g an s DK 53195 and DK 52740)
3037
E ec s o Selec i e and Non-Selec i e COX-1 and COX-2 Inhibi o s on Ch onic
Gas ic Ulce
Healing
Ca alina Ala con de La Las a, Be ina Be engue , Vi ginia Mo il a, Ca men La Case,
Pha macology Dep , Uni o Se ille, Se ille Spain; Juan Manuel He e las, Diges Se ice,
Vi gen Maca ena Hosp, Se ille Spain; Ma ia Jose Ma in, Pha macology Dep , Uni o
Se ille, Se ille Spain
Backg ound. P os aglandins (PGs) de i ed om he inducible cyclooxigenase COX-2, play an
impo an ole in gas ic ulce healing. The ques ion is, whe he he new NSAID, which inhibi
p edominan ly COX-2, a e eally sa e in he long- e m use in condi ions o a p e iously
es ablished ch onic ulce . Aim. To asses he di e en healing p ocess and PGs le els, leukocy e
in il a ion and COX exp ession, using classical NSAID, in compa ison wi h he new selec i e
COX-2 inhibi o celecoxib (CLX), 0.35 mg/Kg, Ma e ials and Me hods. A ch onic ulce was
induced in a s by injec ion o 5% ace ic acid in o he gas ic su ise osa. Animals we e ea ed
wice daily (i.g.) du ing 8 and 14 days wi h ollowing: ehicle, pi oxicam (PRX), 0.35 mg/Kg,
me amizol (MET), 33 mg/Kg, and CI.X, 1.8 mg/Kg. Mac oscopic ulce index (Ul), myelope oxi-
dace ac i i y (MPO) as an index o neu ophil in il a ion and PGE2 con en , we e measu ed
a he ulce si e as well as in mucosa a ound he ulce c a e . His ology and immuno eac i i y
o COX-1 and COX-2 we e also s udied. Resul s. All NSAID delayed ulce healing, al hough
he e whe e no signi ican di e ences in UI. MPO was signi ican ly highe a he ulce si e
han in he in ac mucosa a e ea men s (p<O.O01). PRX signi ican ly inc eased MPO ac i i y
a he ulce si e s sham and con ol (p<O.01). On he con a y CLX dec eased he enzyma ic
ac i i y (p<O.05). PGE~ con en inc eased signi ican ly a he ulce si e (p<O.01) s in ac
mucosa o he ulce a ed con ol. All NSAID supp essed FGE2
gene a ion
in ulce a ed and non-
ulce a ed issues. His ological examina ion showed no impo an di e ences be ween bo h
d ugs bu con i med high neu ophil in il a ion in case o PRX. COX-1 was exp essed p edomi-
nan ly in he neck egion and bo om o gas ic glands and he e we e no any di e ences
be ween g oups. COX-2 was exp essed mainly in egions o maximal epai ac i i y a he
ulce base and supe icial mucous cells and mucous cells o he o eoles. COX-2 exp ession
was ma kedly s onge wi h PRX and con ol han which CLX. Conclusions. Selec i e and
non-selec i e NSAID delay ulce healing, wich may be ela ed wi h a descen in PGE2 con en
and di e en beha iou in COX-1 and COX-2 exp ession. In lamma o y esponses in case o
PRX may be also implica ed.
m
ODes Dasldc
Acid
Sec e ion Accele a e
NSAIO-lnducnd Tissue
Pe exida ion In Ra
S omadl?
Hi o umi Ma soi, I oe Making, Yumiko Nagano, Yasushi Mu a a, Aki a Nakaha a, Naomi
Tanaka, Uni o Tsukube, Tsukuba Japan
BACKGROUND: Gas ic mucosa was pe oxidized du ing NSAID-induced gas ic mucosal
lesions o ma ion, Recen ly, a monoclonal an ibody o a issue p o ein eac ed wi h 4-
hyd oxynononal (4HNE) has become a ailable, which can indica e dis ibu ion o issue pe oxi-
da ion. Using his monoclonal an ibody, we ha e p e iously epo ed ha he deep pa o
he gas ic mucoSa is pa icula ly pe oxidized ollowing NSAID ea men . The pa ie al cells
which a e pa icula ly loca ed in he deep pa o he gas ic mucosa, consume g ea amoun
o oxygen when hey p oduce gas ic juice. This high oxygen consump ion may lead o
enhanced p oduc ion o oxygen adicals and hus inc eased issue pe oxidiza idn in he deep
pa o gas ic mucosa, bu his possibili y has no been es ed a all. AIM: We hus aimed
o elucida e he ela ionship be ween issue pe oxida ion and acid sec e ion a e he NSAID
ea men . Fo his pu pose, we in es iga ed whe he he p e ea men wi h abep azole, one
o a p o~ pump inhibi o (PPI), a ec ed he 4HNE gene a ion induced by NSAID ea men
o no . METHODS: Male Wis a a s we e di ided in o he ollowing 4 g oups: 1) con ol
g oup, 2) indome hacin g oup, 3) abep azole g oup, 4) abep azole and indome hacin g oup.
Each a in bo h he indoma hacin g oup and he abep azole and iodome hacin g oup was
adminis e ed 30 mg kg indome hacin pe us a e 24 hou s as ing. Ra s in bo h he abep azola
g oup and he abep azole and indome hacin g oup was p e ea ed 30 ain be o e he indome h-
acin adminis a ion wi h 10 mg/kg abep azole. Each a s omach was emo ed a ei he O,
15, 30, 60, 180 o 300 mio al e he NSAID ea men . These c yosec ions o hese s omachs
we e immunohis ochamically s ained wi h bo h an i-4HNE an ibody and an i-8(OH)dG an ibody.
RESULTS: 4HNE began o appea in he deep laye o he gas ic mucosa a 30 ain a e he
indome hacin ea men . The a ea o 4HNE ime-sequen ially ex ended o he su ace laye .
The o ma ion o 8(OH)dG appea ed om 180 ain a e he ea men in he same a ea as
4HNE. The p e ea men wi h abep azole signi ican ly inhibi ed 4HNE gene a ion and 8(OH)dG
o ma ion. CONCLUSION: The indome hacin ea men induced issue pe oxida ion as well as
ONA inju y, eapec~ly in he deep pa o , he gas ic mucoca. The p e ea men wi h PPI
signi ican ly inhibi ed he NSAID ea men -induced issue pe oxida ion. These esul s sugges
ha gas~ acid sec e ion accele a es issue pe oxida ion in gas ic mucosa.
3639
6asb4c Damage Induced By Di e en Doses O Indomo hacin In Ra s is
Va iably
Al eclnd By NOS Inhibi ms.
Rica do B. Oli ei a, Ma cellus H L P Souza, Fe nando O. Cunha, Facul y de Medicine de
Rib P a o, Ribei ao P e o B azil
BACKGROUND/OBJECTIVES: Gas oin es inal damage is he majo limi a ion o he use
NSAIOs. Ni ic oxide (NO) is a media o o gas oin es inal mucosal de ence, bu i also
con ibu ed o mucosel inju y in some ci cums ances. The ole o NO in he NSAID-gas min es i-
nal damage was no de ined. We in es iga ed he e ec o inhibi o s o bo h inducible and
cons i u i e NOS on he gas ic damage (GD) induced by di e en doses o indome hacin
(INDO).
MATERIAL AND METHODS: GD was induced by in agas ic ins ila ion o INDO (2.5;
5; 20 and 40mg/Kg) and assessed 1, 3, 6, 12, 24 hs la e . A lesion index (LI) was calcula ed
as he sum o he leng hs o all lesions. Di e en g oups, ecei ed dexame hasone (lmg/Kg),
L-NAME (lOOmg/Kg), N-Ni o-a ginine (lOOmg/Kg), aminoguanidine (lOOmg/Kg), L-A ginine
(1000mg/Kg), D-A ginine (lO00mg/Kg) o L-Lisine (lO00mg/Kg). A e 1 hou , GD was induced
by INDO (5 o 20mg/Kg) and 3 hs la e he animals we e killed and LI was calcula ed.
RESULTS:
INDO caused a
signi ican
and dose-
dependen GD wi h maximal e ec a a dose
o 20 mg/Kg (p<O.O01). GD peaked a 3 hs a e injec ion o INDO (5 o 20mg/Kg) and
e u ned o con ol le els a he end o 24 hs. P e ea men wi h dexame hasone
(LI = 12.0±5.2mm; N = 10), L-NAME (LI = 12.7±3.6mm; N = 10), N-Ni o-A ginine
(LI= 12.3±6.3mm; N=IO), Aminoguanide (LI =11.3±5.3mm; N= O), o L-A ginine
(Ll=6.0±3.1mm; N=IO) bu no D-A ginine (Ll=21.4±6.1mmn; N= 7) o L-Lisine
(Ll=29.4-+ll.5mm; N=5) signi ican ly inhibi ed (P<O.05) he INDO 20 mg/Kg induced
damage (Ll=30.2±4.8mm; N=16). In con as , GD induced by INDO 5mg/Kg
(Ll=7.8±2.2mm; N=15) ended o inc ease by p e ea men by bo h L-NAME
(LI = 11.4_+3.2mm; N = 15) and N-Ni o-a ginine (LI = 17-4.6mm; N = 9), bu he di e ences
did no a ain s a is ical signi icance (p= 0,2 and p=g,36), whe eas he p e ea men wi h
L-A ginine plus L-NAME (LI = 1.4 ±0.1 mm; N = 10) educed signi ican ly (p<O.05) GD induced
by INO0 5mg/Kg
wi h L-NAME (11.4±3.2mm; N =15). Aminoguanidine had no e ec
(LI =6.5±2.2mm; N=5), whe eas L-A ginine (LI = 5.3±2.1mm; N =10), bu no D-A ginine
(LI = 11.7_+5.5mm; N = 5), ended o educe GD induced by iNDO 5mg/Kg (LI = 7.3±2.2mm;
N=15). CONCLUSIONS 1-NOS inhibi o s had a di e en e ec s depending on he dose o
he
INO0. 2- The p e ea men wi h NOS subs a e as well as inhibi o s impai ed gas ic
damage induced by INDO 20 mg/Kg. 3-
The
e ec o L-A gine seems may
be
speci ic, since
because D-A ginine and L-lisine do no cause i . Financial suppo = FAPESP, PRONEX.
A-598