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MALDI-TOF Fingerprinting of Seminal Plasma Lipids in the Study of Human Male Infertility

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Lipids

Abstract

This study proposed lipid fingerprinting of human seminal plasma by mass spectrometry as an analytical method to differentiate biological conditions. For this purpose, we chose infertile men as a model to study specific conditions, namely: high and low seminal plasma lipid peroxidation levels (sub-study 1.1), high and low sperm nuclear DNA fragmentation (sub-study 1.2), and intervention status: before and after subinguinal microsurgical varicocelectomy (study 2). Study 1 included 133 patients, of which 113 were utilized for sub-study 1.1 and 89 for sub-study 1.2. Study 2 included 17 adult men submitted to subinguinal varicocelectomy, before and 90 days after varicocelectomy. Lipids were extracted from seminal plasma and submitted to Matrix-Assisted Laser Desorption Ionization Quadrupole-Time-of-Flight Mass Spectrometry in the positive ionization mode. Spectra were processed using Waters® MassLynx, and MetaboAnalyst online software was used for statistical analyses. For sub-studies 1.1 and 1.2, and study 2, univariate analysis revealed 8, 87 and 34 significant ions, respectively. Multivariate analysis was performed through PCA and PLS-DA. PCA generated 56, 32 and 34 components respectively for each study and these were submitted to logistic regression. A ROC curve was plotted and the area under the curve was equal to 97.4, 92.5 and 96.5 %. PLS-DA generated a list of 19, 24 and 23 VIP ions for sub-studies 1.1 and 1.2, and study 2, respectively. Therefore, this study established the lipid profile and comparison of patterns altered in response to specific biological conditions.

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Abbreviations

CNPq:

National Council of Technological and Scientific Development

DNA:

Deoxyribonucleic acid

DTT:

Dithiothreitol

EDTA:

Ethylenediamine tetraacetic acid

FAPESP:

Sao Paulo Research Foundation

LPO:

Lipid peroxidation

m/z :

Mass-to-charge ratio

MALDI:

Matrix-assisted laser desorption ionization

MALDI-TOF MS:

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry

MDA:

Malondialdehyde

mRNA:

Messenger ribonucleic acid

MS:

Mass spectrometry

OS:

Oxidative stress

PCA:

Principal component analysis

PLS-DA:

Partial least square-discriminant analysis

ROC:

Receiver operating characteristics

SD:

Standard deviation

SPSS:

Statistical package of the social sciences

TBA:

Thiobarbituric acid

TBARS:

Thiobarbituric acid reactive substances

TBE:

Tris, boric acid and EDTA

TCA:

Trichloroacetic acid

UNIFESP:

Sao Paulo Federal University

VIP:

Variable importance in projection

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Acknowledgments

The authors wish to acknowledge National Council of Technological and Scientific Development (CNPq) for providing funding for the research (Process 472941/2012-7) and a scholarship to Ms. Camargo, and Ms. Bruna de Lima. We also thank the Sao Paulo Research Foundation (FAPESP) for providing a scholarship to Ms. Intasqui. The fund providers had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflict of interest

The authors have no conflict of interest to disclose.

Author information

Authors and Affiliations

  1. Department of Surgery, Division of Urology, Human Reproduction Section, Sao Paulo Federal University, R Embau, 231, Sao Paulo, 04039-060, SP, Brazil

    Mariana Camargo, Paula Intasqui, Camila Bruna de Lima, Daniela Antunes Montani, Edson Guimarães Lo Turco & Ricardo Pimenta Bertolla

  2. Hospital Sao Paulo, Sao Paulo, Brazil

    Edson Guimarães Lo Turco & Ricardo Pimenta Bertolla

  3. Department of Animal Reproduction, School of Veterinary Medicine, University of Sao Paulo, Sao Paulo, Brazil

    Marcílio Nichi

  4. Institute of Chemistry, University of Campinas and National Institute of Science and Technology Bioanalytical, Sao Paulo, Brazil

    Eduardo Jorge Pilau & Fabio Cesar Gozzo

  5. Department of Chemistry, Center for Exact Sciences, Maringa State University, Maringa, PR, Brazil

    Eduardo Jorge Pilau

Authors
  1. Mariana Camargo
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  2. Paula Intasqui
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  3. Camila Bruna de Lima
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  4. Daniela Antunes Montani
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  5. Marcílio Nichi
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  6. Eduardo Jorge Pilau
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  7. Fabio Cesar Gozzo
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  8. Edson Guimarães Lo Turco
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  9. Ricardo Pimenta Bertolla
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Corresponding author

Correspondence to Ricardo Pimenta Bertolla.

Electronic supplementary material

Below is the link to the electronic supplementary material.

11745_2014_3922_MOESM1_ESM.pdf

Supplementary material 1 (PDF 112 kb) Supplementary Fig. 1 Example MALDI spectrum for study 2 (Pre and Post-varicocelectomy)

11745_2014_3922_MOESM2_ESM.pdf

Supplementary material 2 (PDF 525 kb) Supplementary Table 1 Significant ions of sub-studies 1.1 and 1.2 and study 2, revealed by Univariate analysis, and their respective lipid category, class, error mass, fold-change

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Camargo, M., Intasqui, P., Bruna de Lima, C. et al. MALDI-TOF Fingerprinting of Seminal Plasma Lipids in the Study of Human Male Infertility. Lipids 49, 943–956 (2014). https://doi.org/10.1007/s11745-014-3922-7

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  • DOI: https://doi.org/10.1007/s11745-014-3922-7

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