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ImmunohIstochemIcal evaluatIon of prolIferatIon, apoptosIs and steroIdogenIc enzymes In the ovary of rats wIth polycystIc ovary rev assoc med Bras 2014; 60(4):349-356 349original articleImmunohistochemical evaluation of proliferation, apoptosis and steroidogenic enzymes in the ovary of rats with polycystic ovary Conflict of interest: noneObjective: to evaluate the immunohistochemical expression of proliferative, apoptotic and steroidogenic enzyme markers in the ovaries of rats with polycystic ovary syndrome (PCOS). Methods: twenty rats were divided into two groups: GCtrl -estrous phase, and PCOS -with polycystic ovaries. The GCtrl animals were subjected to a lighting period from 7 am to 7 pm, while the animals with PCOS group remained with continuous lighting for 60 days. Subsequently, the animals were anesthetized, the ovaries were removed and fixed in 10% formaldehyde, prior to paraffin embedding. Sections were stained using H.E. or subjected to immunohistochemical methods for the detection of Ki-67, cleaved caspase-3, CYP11A1, CYP17A1 and CYP19A1. The results were analyzed using Student's t-test (p < 0,05). Results: morphological results showed evidence of interstitial cells originating from the inner theca cells of degenerating ovarian cysts in PCOS. Immunoexpression of Ki-67 was higher in the granulosa cells in GCtrl, and the theca interna cells in PCOS, while cleaved caspase-3 was higher in granulosa cells of ovarian cysts from PCOS and in the theca interna cells of GCtrl. Immunoreactivity of CYP11A1 in the theca interna, granulosa and interstitial cells was similar between the two groups, while CYP17A1 and CYP19A1 were higher in the granulosa and interstitial cells in the PCOS group. Conclusion: the results indicate that the interstitial cells are derived from the theca interna and that enzymatic changes occur in the theca interna and interstitial cells in ovaries of rats with PCOS, responsible for the high levels of androgens and estradiol.

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We may consider that these structures are clinically relevant findings and, due to the low accuracy of the panoramic radiographs, high-quality tomographic exams should always be asked for presurgical treatment planning.

Il dossier raccoglie i contributi discussi nel corso di una giornata di lavoro dedicata alla rete idrica Ostiense e Portuense. I singoli resoconti fanno parte di un’unica ricerca finalizzata a riassumere i risultati delle scoperte nel territorio, moltiplicatesi negli ultimi anni, e a fornire un primo strumento di lavoro sui problemi tecnici relativi all’approvvigionamento idrico di Ostia, Porto e dei siti del litorale. La raccolta della ricca documentazione grafica, fotografica e antiquaria esistente, confrontata con i ritrovamenti avvenuti sia recentemente che nel secolo scorso, consente l’individuazione di tre grandi linee di trasporto idrico e di posizionarle su planimetria. Mentre gli acquedotti diretti a Porto e al territorio Laurentino, in particolare al Vicus Augustanus Laurentium, presentano un andamento chiaro e regolare, l’acquedotto Ostiense si configura molto più complesso e forse costituito da varie canalizzazioni che nel tempo avrebbero contribuito all’alimentazione idrico di Ostia. Di questo, che al momento si può definire un sistema di acquedotti, vi sono testimonianze già a più di 13 km da Ostia e oltre, fino a giungere alle porte ed all’interno della città antica, ove ricerche recentissime ne hanno messo in luce i collegamenti con il grande Castellum aquae sito presso Porta Romana. Una prima analisi delle caratteristiche idrogeologiche del territorio, consente di avanzare ipotesi sulle opere di captazione che si situano, per Ostia, nella parte alta della valle percorsa dal fosso di Malafede e, per Porto, lungo il fosso della Magliana. Per le zone della costa poste a sud di Ostia si è invece più inclini a pensare ad un sistema di convogliamento, in unico canale, di più cunicoli drenanti, data la presenza di varie sorgenti, ma di piccola portata. Ai resoconti di scavi e scoperte si aggiungono contributi di carattere epigrafico, di cultura materiale e di cartografia storica, con cui si intende riassumere ad oggi lo stato delle ricerche e degli studi, da cui partire per nuovi, successivi approfondimenti.

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SUMMARY OBJECTIVE To evaluate the ovarian effects of melatonin (Mel) in a rat model of polycystic-ovary-syndrome (PCOS) before and after permanent estrus induction. METHODS Thirty-two adult-female rats with regular estrous cycle were equally divided into four groups: 1) GCtrl – at estrous phase. 2) GPCOS - at permanent-estrous phase. 3) GMel1 – treated for 60 days with Mel (0.4 mg/Kg) during permanent estrus induction and 4) GMel2 – rats with PCOS and treated for 60 days with Mel. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 (Casp-3) detections. RESULTS The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in Mel-treated groups, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells. These results were more evident in GMel1. The percentage of Ki-67-positive cells was significantly higher in the Mel-treated groups, mainly in the GMel2, as compared to GPCOS. On the other hand, the percentage of Casp-3-positive cells was significantly lower in granulosa cells of GMel1, whereas it was significantly higher in the interstitial-like cells of GMel2, in comparison to GPCOS. CONCLUSION Melatonin administration prevents the permanent estrus state in the PCOS rat model. This effect is more efficient when melatonin is administered before permanent estrus induction.

The interstitial cells of the rat polycystic ovary probably originate from ovarian cysts due to the degeneration of granulosa cells and differentiation of the internal theca cells. The elevations of serum testosterone and estradiol were probably due to the significant increase in cell activity and in the area occupied by interstitial cells.