Ceramide-1-phosphate (C1P) protects against cyclophosphamide-induced gonadotoxicity in a mouse model
Natalia Pascuali (AR), Leopoldina Scotti (AR), Mariana Di Pietro (AR), Gonzalo Oubiña (AR), María May (AR), Antonio Gómez Muñoz (AR), Dalhia Abramovich (AR), Fernanda Parborell (AR)
[Pascuali] Experimental Medicine and Biology Institute (IByME-CONICET), [Scotti] Experimental Medicine and Biology Institute (IByME-CONICET), [Di Pietro] Experimental Medicine and Biology Institute (IByME-CONICET), [Oubiña] Experimental Medicine and Biology Institute (IByME-CONICET), [May] Instituto de Investigaciones Farmacológicas (ININFA-UBA-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina., [Gómez Muñoz] Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias y Tecnología, Universidad del País Vasco (UPV/EHU), Bilbao, España., [Abramovich] Experimental Medicine and Biology Institute (IByME-CONICET), [Parborell] Experimental Medicine and Biology Institute (IByME-CONICET)
Context: Ovarian dysfunction is a complication of cyclophosphamide (CTX) chemotherapy. Despite advances, available strategies fail to preserve fertility. Sphingolipids like ceramide-1-phosphate (C1P) regulate cell homeostasis. We have shown that C1P, a proangiogenic and anti-apoptotic agent, improves folliculogenesis and endothelial area compared to CTX-treated mice ovaries. Objective: To study the effect of C1P on primordial follicles (PrF), apoptosis and vascular stability in CTX-treated mice ovaries, as well as on endometrium morphology. Methods: 8-week old F1 (BalbC x C57/BL6) female mice were allotted into 3 groups (n=6/group) and received a single i.p. injection of either saline (control) or 75 mg/kg of CTX. CTX mice received intrabursal injection of vehicle (CTX) or C1P (CTX+C1P: 10ul/ovary; 1 mM). On day 14, their ovaries and uteruses were removed and fixed in Bouin’s solution or lysated for protein analysis by Western Blot. Paraffin-embedded ovarian slides were immunostained for DDX4 (PrF marker), cleaved-caspase-3 and α-actin (periendothelial marker) by immunohistochemistry (IHC). Main outcome measures: %PrF/ovary, periendothelial area, % positive cleaved-caspase-3 nuclei, Sphingomyelinase (Spm), Bax and BCLX-L protein expression. Results: Local C1P administration in CTX-treated ovaries restored the %PrF and decreased the % positive cleaved-caspase-3 cells and Spm expression. C1P also increased Bcl-XL:Bax ratio in CTX-treated ovaries. IHC for α-actin showed less periendothelial area in CTX-treated ovaries, which was restored by C1P administration. CTX-treated mice presented damaged endometria, while CTX+C1P uteruses showed normal architecture. Conclusions: C1P preserves ovarian function in CTX-treated mice by protecting PrF reserve, decreasing apoptosis and Spm expression and improving vascular stability. C1P might serve as ovarian cytoprotector in cancer.