Pro-inflammatory agents nitric oxide and TNF-αarrest GC-1 spermatogonia cell cycle through different mechanism

MARÍA SOFÍA AMARILLA; MARÍA EUGENIA FERREIRO; LEILANE GLIENKE; LUCAS NICOLAS GONZÁLEZ; PATRICIA VERÓNICA JACOBO; CRISTIAN SOBARZO; ANDREA DE LAURENTIIS; MARÍA JIMENA FERRARIS; MARÍA SUSANA THEAS

Mar del Plata

Congreso; LVIII Reunión Anual de la SAIC; 2019

Sociedad Argentina de Investigaciones clínicas

Nitric oxide (NO) and Tumor Necrosis Factor alpha (TNFα) are pro-inflammatory agents able to interferewith cell cycle. Experimental autoimmune orchitis (EAO) is a model of chronic inflammation associatedto infertility. In EAO high levels of NO and TNFα are produced by testicular macrophages and pre-meiotic germ cells (spermatogonia and pre-leptotene spermatocytes) proliferation is reduced. We proposethat NO and TNFα arrest spermatogonial cell cycle in EAO. To evaluate this hypothesis we explored theeffect of DETA-NO (a NO donor) and TNFα on cell cycle and death on GC-1 spermatogonia cell line byflow cytometry. Both TNF-α (50ng/ml) and DETA-NO (2.0mM), significantly increased the percentageof 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 24and 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 andapoptosis while NAC did not modify TNFα action. DETA-NO induced GC-1 cell cycle arrest andapoptosis was reverted after DETA-NO withdrawal unlike TNFα.Our results showed that NO and TNFα arrest the cell cycle of GC-1 spermatogonia. While NO effect onGC-1 cells was mediated by oxidative stress and reversible, TNFα action was independent of oxidativestress and irreversible. We conclude that NO and TNFα might control spermatogonial cycle progressionthrough different pathways in the testis under chronic inflammation.