SECUNDARE ALE LEFLUNOMIDULUI (ARAVA) ÎN ARTRITA CRONICĂ INDUSĂ EXPERIMENTAL
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1 EFECTELE SECUNDARE ALE LEFLUNOMIDULUI (ARAVA) ÎN ARTRITA CRONICĂ INDUSĂ EXPERIMENTAL Mihaela Chicu 1, E. Bitere 2, Camelia Mutică 3, Magda Bădescu 1, Gioconda Dobrescu 4 1- DISCIPLINA DE FIZIOPATOLOGIE, UNIVERSITATEA DE MEDICINĂ ŞI FARMACIE GR.T.POPA IAŞI; 2- INSTITUTUL DE CARDIOLOGIE G. GEORGESCU, IAŞI; 3- POLICLINICA CFR GALAŢI; 4- DISCIPLINA DE HISTOLOGIE, UNIVERSITATEA DE MEDICINĂ ŞI FARMACIE GR.T.POPA IAŞI, ROMÂNIA SIDE EFFECTS OF LEFLUNOMIDE (ARAVA) IN INDUCED CHRONICAL ARTHRITIS Abstract. Introduction. Pharmacology therapy of chronic inflammatory arthritis has modified in the last year, by introducing of a new substances which can the delay the evaluation of disease and maintaining functions and joint structure. Leflunomide is one of this drugs which acting by blocking lymphocyte proliferation and dihidroorotat dehidrogenase action. Leflunomide inhibit also IL 1β, TNFα, NO and MMP-3 production. Material and methods. We have induced an inflammatory chronic arthritis on Wistar female rats, young and adults, using carrageenan 1% reagent, followed by Leflunomide administration (20 mg/kgc). The experiment period was about 8 weeks. At the ending of the experiment we have drawing heart, liver and kidney fragments which were analyzed on electronic microscopy. Results. Leflunomide was responsible by toxic effects on heart, liver and kidney, but improve the evolution of chronic inflammatory arthritis. Conclusions. Leflunomide delay the evolution of chronic inflammatory arthritis, but doesn t hilling this diseases. Key words: arthritis, Leflunomide, heart, liver, kidney Rezumat. Introducere. Terapia farmacologică a artritelor inflamatorii cronice s-a modificat mult în ultimii ani prin introducerea unor substanţe cu potenţial de influenţare a evoluţiei bolii, în sensul că, pe lângă ameliorarea simptomelor determină şi o reducere a gradului de deteriorare articulară, cu păstrarea structurii şi funcţiei acesteia. Din grupa acestor droguri face parte şi Leflunomidul, al cărui mecanism de acţiune se bazează pe blocarea proliferării limfocitelor prin inhibarea unei enzime dihidroorotatdehidrogenaza, cu rol în sinteza de novo a bazelor pirimidinice. Leflunomidul inhibă producţia de IL 1β, TNFα, NO şi MMP-3 în culturile celular sinoviale umane activate. Material şi Metodă. Pornind de la aceste aspecte, am realizat un model experimental de artrită inflamatorie cronică pe şobolani femele rasa Wistar, tinere şi adulte, la care am administrat Leflunomid în doză de 20 mg/kgc prin gavaj. Durata experimentului a fost de 8 săptămâni, iar în această perioadă animalele au primit hrană standard şi au fost corect hidratate. La finalul experimentului s-au recoltat fragmente de cord, ficat, rinichi, care au fost supuse examenului anatomopatologic. Rezultate. Afectarea hepatică a fost deosebit de intensă la grupul de animale tratate cu Leflunomid (distrofie lipidică, infiltrate inflamatorii abundente în spaţiul port); afectarea cardiacă a confirmat pentru prima dată în literatura de specialitate efectul toxic asupra cordului (fibre miocardice necrozate la coloraţia Lie), iar la nivel renal prezenţa de numeroase infiltrate inflamatorii cronice. Concluzie. Deşi a întârziat evoluţia fenomenului de distrucţie articulară, Leflunomidul se va administra cu prudenţă la pacienţii cu leziuni hepatice, cardiovasculare şi renale. Cuvinte cheie: artrită, Leflunomid, cord, ficat, rinichi
2 Introducere Terapia farmacologică a artritelor inflamatorii cronice s-a modificat mult în ultimii ani prin introducerea unor substanţe cu potenţial de influenţare a evoluţiei bolii, în sensul că, pe lângă ameliorarea simptomelor determină şi o reducere a gradului de deteriorare articulară, cu păstrarea structurii şi funcţiei acesteia. Din grupa acestor droguri face parte şi Leflunomidul, al cărui mecanism de acţiune se bazează pe blocarea proliferării limfocitelor prin inhibarea unei enzime dihidroorotatdehidrogenaza, cu rol în sinteza de novo a bazelor pirimidinice. Leflunomidul inhibă producţia de IL 1β, TNFα, NO şi matrix metalo protease (MMP-3) în culturile celular sinoviale umane activate (1, 2). Material şi metode Studiul experimental a constat in inducerea unei inflamaţii cronice articulare la animalele de laborator, în acest caz la şobolani rasa Wistar de sex feminin si urmărirea in timp a efectelor Leflunomidului asupra evoluţiei clinice a acesteia. Raţiunea alegerii acestui medicament a fost aceea că administrarea s-a se însoţeşte de bune rezultate clinice susţinute în timp, dar cu o serie de reacţii adverse ca anemia severă, infecţiile, leziunile hepatice, risc de dezvoltare a malignităţilor, fenomene observate şi de noi în cursul acestui experiment. Am preferat şobolani femele, întrucât frecvenţa fenomenelor inflamatorii reumatismale este mult mai mare la sexul feminin. Pornind de la aceste aspecte, am realizat un model experimental de artrită inflamatorie cronică pe şobolani femele rasa Wistar, tinere şi adulte, cu soluţie carrageenan 1% în articulaţia tibiofemurală, urmat de administrarea zilnică de Leflunomid în doză de 20 mg/kgc prin gavaj, timp de 8 săptămâni. Animalele au primit hrană standard şi au fost corect hidratate. Pentru a valida rezultatele experimentului, la finalul acestuia, am recoltat fragmente din articulaţia tibiofemurală, pe care am investigat-o în microscopia optică după prelucrare (decalcifiere, colorare), respectiv fragmente de organe (ficat, rinichi, cord), în scopul evaluării eventualelor reacţii adverse ale substanţelor administrate (carrageenan, Leflunomid) asupra acestora. Coşoraţiile utilizate în studiul nostru au fost: hematoxilina-eozina (HE), Van Gieson, Gőmőri şi Coloraţia LIE pentru fibrele miocardice. Rezultate Secţiunile au fost examinate la microscopul OLIMPUS de la Disciplina de Dermatologie a UMF Iaşi, de către doamna Academician Prof. Dr. Docent Gioconda Dobrescu. La animalele din lotul martor aspectul articulaţiei tibiofemurale a fost normal: spaţiul articular a fost liber, suprafaţa cartilagiului articular a fost intactă, cele două zone de cartilaj (nemineralizat şi mineralizat) de dimensiuni normale. Placa de os subcondral separă zona de măduvă osoasă de cartilagiul mineralizat (fig. 1, fig. 2). Fig. 1 Şobolan martor tânăr 61
3 Fig. 2.Şobolan martor adult La lotul de animale adulte la care s-a produs artrita experimentală am constatat leziuni de artrită cronică erozivă la toate animalele. Cele două zone de cartilaj: nemineralizat şi mineralizat au apărut subţiate fie pe un versant fie pe ambele, sau cele mai frecvent dezorganizate, erodate şi înlocuite de panusul inflamator. Placa osoasă subcondrală redusă, iar în cavitatea articulară abundent panus în inflamator. Am constatat şi zone de rezorbţie osoasă la nivelul plăcii osoase subcondrale. Cantitatea de panus intraarticular a fost variabilă, fie a ocupat parţial cavitatea articulară, fie a obliterat cavitatea articulară. Într-un caz panusul a ajuns să erodeze osul trabecular, care prezenta leziuni de liză osoasă şi chiar început de fibroză. Sinovia îngroşată, hipertrofiată, iar ţesutul adipos subjacent a fost invadat de celule inflamatorii (fig. 3). La lotul de şobolani adulţi cu artrită, trataţi cu Leflunomid s-au constatat leziuni artritice mai reduse, cu puţin panus intraarticular şi uşoară reducere a zonelor cartilaginoase. Placa osoasă subcondrală este mult diminuată (fig. 4). Fig. 3: Şobolan adult cu artrită Fig. 4: Şobolan adult cu artrită tratat În lotul de animale tinere la care s-au produs leziunile de artrită modificările morfologice au fost mai exuberante, cu importante leziuni de dezorganizare a cartilagiului şi abundent panus intraarticular, la nivelul căruia au apărut fie zone de necroză fibrinoidă, fie infiltrate inflamatorii importante. Cartilajul articular dezorganizat a fost înlocuit de panusul inflamator. Osul trabecular cu zone de osteoliză, în jurul cărora se constituie o reacţie fibroasă. Şi în acest lot au fost prezente leziunile de îngroşare şi proliferare a sinovialei, cu reacţie inflamatorie subjacentă (fig. 5). La lotul de şobolani tineri cu artrită, trataţi cu Leflunomid se constată o reducere semnificativă a panusului intraarticular (fig. 6). 62
4 Fig. 5: Şobolan tânăr cu artrită Fig. 8. Fibre miocardice necrozate, Lie+ nodular interstiţial - ficat La majoritatea animalelor examinate s-a constatat la nivelul ficatului leziuni de distrofie lipidică, micro- şi macro-veziculară şi infiltrate inflamatorii în spaţiile porte (fig. 9, 10). Fig. 6: Şobolan tânăr cu artrită tratat În ceea ce priveşte efectele Leflunomidului asupra principalelor organe interne (cord, ficat, rinichi) am constatat următoarele: - cord se constată tumefierea şi omogenizarea a fibrelor miocardice cu infiltrat inflamator nodular interstiţial (fig. 7) şi prezenţa fibrelor miocardice necrozate la coloraţia Lie+ (fig. 8). Fig. 9. Ficat distrofie lipidică Fig. 10. Infiltrate inflamatorii în spaţiile porte Fig. 7. Cord infiltrat inflamator - rinichi, inconstant, am observat prezenţa infiltratelor inflamatorii cronice (fig. 11). 63
5 Fig. 11. Rinichi - infiltrate inflamatorii cronice Concluzii 1. Experimentul a evidenţiat efectele toxice viscerale ale Leflunomidului asupra ficatului, cordului şi rinichiului. 2. Afectarea hepatica a fost deosebit de intensa la loturile de animale tratate cu Leflunomid confirmând datele din literatura de specialitate privind hepatotoxicitatea acestuia. 3. Pentru prima dată în literatura de specialitate am evidenţiat efectul toxic al Leflunomidului asupra cordului. 4. Examenul anatomopatologic al rinichiului a decelat modificări determinate atât prin administrarea de Leflunomid cât şi prin carrageenan. 5. Decesele survenite în lotul de animale tratate cu Leflunomid s-au datorat fenomenelor de insuficienţă hepatică acută, hemoragiilor, infecţiilor locale, diminuării apetitului şi consecutiv scăderii în greutate. 6. Deşi a întârziat evoluţia fenomenului de distrucţie articulară, Leflunomidul se va administra cu prudenţă la pacienţii cu leziuni hepatice, cardiovasculare şi renale. Bibliografie Codreanu C: Concepţii moderne în terapia poliartritei reumatoide, Revista de Reumatologie nr. 1, volum VI/1998. Zvaifler N. - Ethiology and pathogenesis of rheumatoid arthritis in Arthritis and allied conditions, 11th Ed.- McCarty, 1986, pg
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