Effects of n-6 and n-3 polyunsaturated acid-rich soybean phosphatidylcholine on membrane lipid profile and cryotolerance of human sperm.
OBJECTIVE
To study the effects of n-6 and n-3 polyunsaturated acid-rich soybean phosphatidylcholine (soy-PC) on sperm cryotolerance with regard to sperm membrane lipid profile, membrane surface integrity, and routine semen parameters.


METHODS
Experimental study.


METHODS
University-affiliated tertiary hospital.


METHODS
A total of 20 normospermic fertile men.


METHODS
Semen samples examined for differences in semen parameters, sperm membrane lipid profile, and plasma membrane surface both before and after cryopreservation using basic freezing medium with N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid (TES) and tris-(hydroxymethyl)-aminomethane (TRIS) supplemented with purified soy-PC (TEST-PC) or egg yolk (TEST-Y), both alone or in association (TEST-Y-PC).


METHODS
Conventional semen parameters and membrane lipid profile by matrix-assisted laser/desorption ionization mass spectrometry (MALDI-MS).


RESULTS
Postthaw sperm cell motility, vitality, and morphology parameters were similar for soy-PC (TEST-PC) and egg yolk (TEST-Y) cryoprotectants. However, sperm exposed to TEST-Y-PC presented better kinetic parameters, which were similar to the original quality of the fresh semen. Human sperm MALDI-MS lipid profiles revealed that the relative abundance of glycerophospholipids of m/z 760.44 [PC (34:1)+H]+, 781.55 [SM (20:0) +Na]+, 784.55 [PC (36:3) +H]+, 806.64 [PC (38:6) +H]+, 807.64 [SM (22:1) +Na]+, and 809.64 [SM (22:0) +Na]+ increased in soy-PC samples (TEST-PC). Nonetheless, only one lipid (m/z 781.55, [SM (20:0) +Na]+) statistically significantly changed when sperm was cryopreserved in TEST-Y-PC.


CONCLUSIONS
Sphingomyelin was defined as a prospective biomarker of soy-PC treatment, and it could be related to the positive cryoprotective effects of soy-PC in human sperm, opening new perspectives to design of a more efficient synthetic cryoprotectant medium containing purified egg yolk biomolecules combined with soy-PC.
Department of Obstetrics and Gynecology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.
Alessandra A
AA
Vireque
Metabolite Profiling Facility, Bindley Bioscience Center, Purdue University, West Lafayette, Indiana.
Alessandra
A
Tata
Department of Obstetrics and Gynecology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.
Oswaldo F L L O
OF
Silva
Department of Surgery, Division of Urology, Human Reproduction Section, São Paulo Federal University, São Paulo, Brazil.
Edson G
EG
LoTurco
Department of Surgery, Division of Urology, Human Reproduction Section, São Paulo Federal University, São Paulo, Brazil.
Augusto
A
Azzolini
Metabolite Profiling Facility, Bindley Bioscience Center, Purdue University, West Lafayette, Indiana.
Christina R
CR
Ferreira
Department of Obstetrics and Gynecology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.
Marilda H Y
MH
Dantas
Department of Obstetrics and Gynecology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil; National Institute of Hormones and Woman's Health, CNPq, Porto Alegre, Brazil.
Rui A
RA
Ferriani
Department of Obstetrics and Gynecology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil; National Institute of Hormones and Woman's Health, CNPq, Porto Alegre, Brazil. Electronic address: romareis@fmrp.usp.br.
Rosana M
RM
Reis
D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D003080:Cold Temperature / Q000009:adverse effects*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D004530:Egg Yolk / Q000737:chemistry; D015525:Fatty Acids, Omega-3 / Q000302:isolation & purification / Q000494:pharmacology*; D043371:Fatty Acids, Omega-6 / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism*; D008823:Micelles; D008855:Microscopy, Electron, Scanning; D010713:Phosphatidylcholines / Q000302:isolation & purification / Q000494:pharmacology*; D013025:Glycine max / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D013109:Sphingomyelins / Q000302:isolation & purification / Q000494:pharmacology
D003451:Cryoprotective Agents; D015525:Fatty Acids, Omega-3; D043371:Fatty Acids, Omega-6; D008563:Membrane Lipids; D008823:Micelles; D010713:Phosphatidylcholines; D013109:Sphingomyelins
Lα-Phosphatidylcholine; lipid profile; plasma membrane; sperm cryopreservation