PRO-INFLAMMATORY AGENTS NITRIC OXIDE AND TNF ALPHA ARREST GC-1 SPERMATOGONIA CELL CYCLE THROUGH DIFFERENT MECHANISM

LEILANE GLIENKE; CRISTIAN MARCELO SOBARZO ALVAREZ; MARÍA SUSANA THEAS; MARÍA SOFÍA AMARILLA ; CANDELA ROCÍO GONZÁLEZ; ANDREA DE LAURENTIIS; MARIA EUGENIA FERREIRO; PATRICIA VERÓNICA JACOBO; MARÍA JIMENA FERRARIS

Mar del Plata

Congreso; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2019

SAIC-SAFE-SAB-SAP-NANOMED-ar

Nitric oxide (NO) and tumor necrosis factor alpha (TNF alpha) are pro-inflammatory agents able to interfere with cell cycle. Experimental autoimmune orchitis (EAO) is a model of chronic inflammation associated to infertility. In EAO high levels of NO and TNF alpha are produced by testicular macrophages and pre-meiotic germ cells (spermatogonia and pre-leptotene spermatocytes) proliferation is reduced. We propose that NO and TNF alpha arrest spermatogonial cell cycle in EAO. To evaluate this hypothesis, we explored the effect of DETA-NO (a NO donor) and TNF alpha on cell cycle and death on GC-1 spermatogonia cell line by flow cytometry. Both TNF alpha (50 ng/ml) and DETA-NO (2.0 mM), significantly increased the percentage of GC-1 cells in the S-phase and significantly reduced the percentage in the G1-phase of the cell cycle (propidium iodide incorporation, IP) also inducing cell apoptosis (Annexin V-FITC-IP assay) after 24 and 18 h of incubation respectively. Pre-incubation of GC-1 cells with a general antioxidant, N-acetyl-L-cysteine (NAC, 2.5 and 5.0 mM) significantly reduced DETA-NO effect on cell cycle arrest and apoptosis while NAC did not modify TNF alpha action. DETA-NO induced GC-1 cell cycle arrest and apoptosis was reverted after DETA-NO withdrawal unlike TNF alpha.