APOPTOSIS IN LONG TERM DIABETIC MOUSE UTERUS: AN ANALYSIS OF INTRINSIC AND EXTRINSIC PATHWAY OF APOPTOSIS

MEILERMAN ABUELAFIA ANALIA.; FRAUNHOFFER NICOLAS A.; ROMAN KAREN; D´AMICO PAULA; BLANCO LILA; STURLA CECILIA; DE GREGORIO GUIDO; VITULLO ALFREDO D.; BARRIOS MARCELA

Mar del Plata

Congreso; LXI Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2016

Sociedad Argentina de Investigación Clínica

Type I diabetes mellitus (T1D) accounts for about 10% ofall cases of diabetes. It is a multifactorial autoinmune diseasefor which susceptibility is determined by genetic, environmentaland immunological factors. Emerging evidence has shown thatdiabetic mice uterus presented severe atrophy, glands reduction,fibrosis and destruction of smooth muscle cells. These changeswould be related with an increase of apoptosis. The objectiveof this study was to analyze the balance between the apoptosispathways in diabetic mice uterus. 25 BALB/c female mice (age30 days; 15 diabetic and 10 controls) were used in this assay.To generate a T1D model, female mice received five injectionsof streptozotocin at a dose of 60 mg/kg, during 5 consecutivedays. Animals were selected as diabetic when glucose level was>200 mg/dl. Weight and food intake was recorded weekly both indiabetic and control animals. Three diabetic and 2 control animalswere sacrificed at day 15, 20, 40, 70 and 80 post-treatment. Theuteri were removed and weighed; one part was fixed in 4% PFAfor immunohistochemistry and immunofluorescence and anotherone was frozen at -80 °C for Western Blot analysis. The proteinsstudied were: Bax, Bcl-2, cleaved Bid, Fas/Fas-L system, cleavedcaspase 8 and active caspase 3. Bcl-2 expression was constantat all time-points, with no differences between control and diabeticmice. Bax and extrinsic apoptosis pathway markers (Fas, Fas-L,cleaved Bid and cleaved caspase 8) showed strong cytoplasmaticimmunostaining in endometrial glands and smooth muscle cell ofdiabetic mice uterus at all time-points analyzed. Active caspase 3was positive in glands and smooth muscle cells of diabetic miceuterus with constant levels of expression in all time-points. I n conclusionour results show that during the streptozotocin - inducedT1D, both intrinsic and extrinsic apoptosis pathway are involved inthe endometrial degeneration by promoting glandular and smoothmuscle cell death.