Lipidomics analysis of the vaginal discharge in women with vulvovaginal candidiasis and cytolytic vaginosis

• Grant number: 16/18850-9
• Support type: Scholarships in Brazil - Master
• Effective date (Start): December 01, 2016
• Effective date (End): November 30, 2017
• Field of knowledge: Health Sciences - Medicine
• Principal researcher: Paulo César Giraldo
• Grantee: José Marcos Sanches Junior
• Home Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Background: Vaginal discharge is a very common condition that affects women. Vulvovaginal candisiasis (VVC) and cytolytic vaginosis (CV) are gynecological condition with symptoms that include vaginal itching, burning, abnormal discharge, dysuria and dyspareunia. Although the symptoms are quite similar, the vaginal microenvironment and amount of lactobacillus, inflammatory response, and treatment are completely different. The advances in mass spectrometry techniques and bioinformatics have been improved with the need for the understanding the health and disease process, using the lipidomics as a great tool to characterize potential biomarkers for many diseases. Objective: Characterize the lipids in the vaginal discharge in women with VVC and CV as biomarkers, relating with the physiology, physiopathology and metabolism of these conditions is the aim of this work. Material and methods: A cross-sectional study with 24 women, non-pregnant, pre-menopausal, aged between 18 to 44 years was conducted in the Outpatient Clinic of Genital Tract Infections of the Campinas State University Women's Hospital, Campinas, Brazil. All women provided written informed consent, approved by the Ethics and Research Committee, CAAE:60648016.8.0000.5404. All women provided written informed consent, approved by the Ethics and Research Committee, CAAE:60648016.8.0000.5404. All women were interviewed, had an anamnesis and underwent a speculum-based gynecological examination. After properly diagnosis, the patients were placed in three groups: VVC, CV or health women. The samples collected were resuspended in deionized water following the addition of chloroform HPLC grade and of methanol, vortex and centrifuged. All samples were dried using SpeedVac and kept frozen at -80oC until analysis. Chromatographic separation was performed on an Agilent1290-Infinity ultra-high performance liquid chromatography system, and a C-18 column was also utilized. The data were obtained on the positive and negative mode, by the software MassHunter Qualitative (Agilent). All metabolites were evaluated for mass error (d5 ppm), conclusive isotopic and fragmentation pattern and plausible retention time, characterized as description by The Human Metabolome Database, XCMS and Lipid Maps. Multivariate data analysis techniques were applied to process the acquired data. The segregation between groups was analyzed by PCA, and PLS-DA. Results: The PCA analysis showed an important separation which characterized the metabolites difference between the three groups. Selection of potential biomarkers provided by the PLS-DA is the result of the VIP (Variable Influence on Projection), finding a total of 38 lipids. The main biomarkers for VVC associated with inflammatory process were prostaglandins and O-Adipoylcarnitine. Phosphatidic acids and 13S-HpOTrE fatty acid could increase the signals for inflammation and pain in VCC. The physiopathology of CV is described for the lyse of the vaginal epithelium and lactobacilli overgrowing which have 1-(11Z-docosenoyl)-glycero-3-phosphate, and 5-Aminopentanoic acid as biomarkers, respectively. Also, 1-oleoyl-cyclic phosphatidic acid and Palmitoleic acid are present in injured tissue, possibly playing a role of recovering the vaginal epithelium in CV. Some lipids are also related with oxidative stress and apoptosis, especially in CV that include phosphatidylserines, N-(tetradecanoyl)-sphinganine and Glycochenodeoxycholic acid 7-sulfate. Conclusion: This work has a very important role to elucidate the metabolic mechanisms of the lipids in VVC and CV that could relate with their biofunction and pathways in physiopathology. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SANCHES, JOSE MARCOS; GIRALDO, PAULO CESAR; AMARAL, ROSE; EBERLIN, MARCOS NOGUEIRA; MARQUES, LYGIA AZEVEDO; MIGLIORINI, ISABEL; NAKAHIRA, MARCEL; MARINUS BIELEVELD, MICHEL JAN; DISCACCIATI, MICHELLE GARCIA. Vaginal lipidomics of women with vulvovaginal candidiasis and cytolytic vaginosis: A non-targeted LC-MS pilot study. PLoS One, v. 13, n. 8 AUG 22 2018. Web of Science Citations: 0.