LONG-TERM BROMOPRIDE INTRODUCTION TREATMENT INDUCES MORPHOLOGICAL CHANGES IN FEMALE RATS Bromopride CN - (diethylaminoethyl) - 2-rne t hoxy-4 -5- b r omobenzami ne ) is a widely used drug in gastroenterologic clinic, not only because of its central and peripheral actions on gastrointesti­ LUCIRNO FREITAS FELICIO nal motility btjl also for its antismetic Professor Assistente properties. Faculdade de Medicina Veterinária e Long-term treatments are frequently Zootecnia da U5P performed with this drug. It is known that bromopride acts on the central nervous system blocking dopaminergic IOERCIC LUIZ 5INHÜRINI receptors (1,4), leading to an increase Professor Assistente Doutor in serum prolactin levels through recep­ Faculdade de Medicina Veterinária e tors blockage at the tubero-infundibuI ar Zootecnia da U5P dopaminergic system (6). This increase in prolactin levels is an undesirable side effect in patients chronically JORO PALERMO NETO treated with this drug (6). Professor Titular In this report it is presented the Faculdads de Medicina Veterinária e assessment of long-term bromopride Zootecnia da USP treatment on the morphology of mammary glands, ovaries, uterus and fallopian tubes of rats. ANTONI A GLADYS NR5ELLO Professor Rssociado Faculdade de Ciências Médicas da MRTERIRL RND METHODS Santa Casa

Rdult virgin female rats (Wistar .origin) aging 50 days at the beginning of the treatment were used in this study. Daily vaginal smears were sampled to monitor estrous cyclicity and 20 animals showing two consecutive 4-day- estrous cycle were used. FELICIO, L.F.; SINHORINI, I.L.; PALERMO Three rats were maintained per each NETO, J-i NASELLO, A.G, Long-term bro- cage under controlled light (12h light, nopride treatment induces morphological 12h dark; lights off at 7:00h p.m.) and changes in female rats. Rev. Fac. Med. temperature (22-23°C) conditions. Food Vet. Zootec. Univ. S. Paulo, 25(1): 117- and water were available ad lib. 121,1988. Pure bromopride powder generously provieded by ROCHE was dissolved in 0.2 5UMMARY : There were morphoIogica I y examined M monopotassic phosphate solution anJ pH tha ovaries, uterus, fallopian tubes and was adjusted to 5.0 with 0.5 N mammary glands of adult female rats, that hyd roch loric acid. were long-term treated with bromopride. Half of these animals received 60.0 Large corpus luteum, uterus in a secretory mg/kg of drug subcutaneous I y (5.C.) stage and hipertrophiated mammary gland twice a day during 26 days (experimental were observed in treated animals. These group). The remaining rats were treated narked morphological changes were corre­ only with bromopride solvent in the same lated to the neuroendocrinal effects of the schedule (control group). From the tenth drug, considering what it is able to block tuberoinfundibuI ar dooamine (DA) receptors day of the treatment until the 28th day, lead!..g to an increase in prolactin levels. the vaginal smears were observed once a day at the same time (8:00h a.m.) in order to monitor estrous cyclicity. The UNITERME: Bromopride; Oestrous cycle, animals were sacrificed twenty four diestrous; Prolactin hours after the last injection, and the 118 FELICIO. L.F. et alii . Long-term bromopride treatment induces morphological changes in female rats

uterus, ovaries, fallopian tubes and tological changes mentioned are histolo­ mammary glands were removed for his to lo­ gical correlates of the diestrous gical examination. Rf ter removal, tis­ observed in the vaginal smears. sues were immediatly immersed in Bouin Bromopride is a DO antagonist fixative, included in paraffin, cut (1,4). Thus, this kind of drug is able (thickness: Sum) and stained with not only to increase serum levels of PRL hematoxylin and eosin for ordinary opti­ but also to induce diestrous in rats (5) cal microscopic observation. since DO seems to be one of the The tissues were examined by one of prolactin inhibiting factors (2,3). Pro­ the authors, who previously knew nothing bably this is the reason fjr the rise in about the treatment the animals had PRL levels in healthy women and after received. long-term bromopride treatnent (6). The histological changes observed i r> i.ammary glands are the typical ones of increased PRL levels. On elevated PRL concentra­ RESULTS tions have profound effects on reproduc­ tive cyclicity. Apparently PRL disrupts essential preovulatory neurochemical events in selected brain areas involved The vaginal smears, monitored dur­ in the regulation of'LHRH as well as the ing 18 days had shown that the animals ovarian steroid secretion patterns that of experimental group were permanently accompany the estrous cycle (7,3). in diestrous. On the contrary, rats of The ovsrian changes observed are in control group had normal cyclicity. agreement with the hormonal changes As demonstrated histologically in described by Wise (8) and the uterine Fig. 10 the ovaries of the control rats histological features were the ones showed follicles in different develop­ expected in this kind of ovarian func­ mental stages and corpus luteum corre­ tional modification. Possibly it seems sponding to a normal cycle. In the expe­ that drug-induced ovarian modifications rimental ones, the follicles were not may be the cause of the menstrual disor­ observed and the corpus luteum were well ders observed in patients treated with developed, presenting proliferation of other ortopramides such as clebopride, granulous cell layer, amorphous m’toclopramide and sulpiride (S). intercellular substance, better cellular The long-term bromopride treatment organization and connective tissue with induces morphological changes in female many vessels (Fig. 1B). Further, diffe­ rats arising out of the neuroendocrinal rent morphological features were also effects of the drug. Although high doses observed in the uterus of control were presently used, it is necessary to animals according to the cyclic phase have in mind this kind of side effect they presented at the time of their when long-term use of the drug is con­ sacrifice. The rats of the experimental sidered, especially in gastroenteroIngic group were in a diestrous phase, i.e., a clinic. Patients should be aware of same secretory stage. Whereas normal mammary possible undesirable side effects of the glands were observed in the control drug like gynecomastia in men, rats, hypertrophic and edematous glands galactorrhea, menstrual disorders and with lactic secretion in the even infertility in women. ga I actcphorous ducts were found in rats long-term treated with bromopride. Finally, the fallopian tubes of animals of both groups showed no diffe­ rences .

ACKNOWLEDGMENTS

DI5CUSSI0N

This work was partially supported by FAPESP (Proc. 83/0SS3-2). The authors wish to express sincere thanks to ROCHE Long-term bromoprid.. treatment can LABORATORIES for bromopride supply. This induce permanent diestrous in rat. work is part of M.S. dissertation Ovaries showing absense of follicles, presented by L.F. Felicio to the School with well developed corpus luteum, ute­ of Veterinary Medicine of University of rus in a secretory stage and other his­ SoO Paulo

Rev. Fac. Med. Vet- Zootec. Univ. S. Paulo. 25(1): 117-121. 1988. FIGURE 1A — Ovary of a control rat with follicles in different developmental stages (F) and corpus luteum (C) corresponding to a normal cycle (hematoxylin and eosin, x 80)

FIGURE IB — Ovary of a long-term bromopride treated rat without follicles, hypertrophic corpus luteum (C) (hematoxylin and eosin, x 80) FELICIO. L.F. et aiii 121 Long-term bromopnde treatment induces morphological changes in female rats

FELICIO, L.F.; 51 NHOR INI , I. L PALERMO corpos lúteos muito desenvolvidos, úteros NETO, J.; NRSELL-Q, fl.G. Rlteraç3es em f ôr.e secretária e glândulas mamáriar morfológicas em ratas induzidas pelo hipertrofiadas. Estas alteraçS®s morfolô- tratamento prolongado com o bromopride. gicas marcantes estão correlacionadas com Rev. Fac. Med. Vet. Zootsc. Univ. 5. os- efeitos neuroendòcrinos da droga, pois Paulo, 25(1) :l 17-121, 1988. esta è capaz de bloquear os dopaninccep- tores do sistema tuberoinfundibular, aumen­ tando, assim, os niveis séricos de prolac- t i na . RESUMO: Foram examinados ovários, úteros, trompas de falópio e glândulas mamárias, provenientes de ratas tratadas - prolongada- UNITERMQ5: Bromopride; Ciclo estral, dies- mente com o bromopride. Foratn observados tro; Prolactin

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Recebido para publicação em 16/08/67 Rpi ovado part, publicação em 15/03/68

Rev. Fac. Med. Vet. Zootec. Univ. S. Paulo. 25(1): 117-121. 1988.