Urine exosome biomarkers of obesity after Lekhana Basti treatment - Report of a pilot study.

BACKGROUND: Obesity is a rising risk factor for various diseases including cardiovascular diseases and Cancer. The limitations of targeted obesity-treatment approaches employed in the clinic presently underscore the importance of developing integrative management strategies for identification of specific biomarkers of obesity. OBJECTIVES: Given the specificity of exosome/extracellular vesicle (EV) biomarkers, we aimed here to identify the EV biomarkers of Ayurveda treatment - Lekhana Basti - for Obesity. METHODOLOGY: A total of eighteen 24-h urine samples from 6 participants with BMI>30 kg/m(2) were used in this study, collected over 3 time-points during the Lekhana basti (medicated enema for obesity) treatment. Urine EV were isolated using Polyethylene Glycol (PEG). The proteins were resolved by 1-d gel electrophoresis and identified using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and quantified by label-free methods. Significant Protein-Protein Interactions, KEGG pathway analysis and enrichment, functional gene ontology (GO) annotation were identified and shortlisted in comparison to Obesity reference genes from DisGeNET. RESULTS: With UniProt as a reference subsequent to LC-MS/MS-identification, a total of 210 exosome proteins were identified. Seventy-three proteins were overexpressed in pathway enrichment analysis. Further, GO functional annotation identified 15 common proteins involved. Finally, the 8 hub proteins associated with obesity were identified and their differential expression profile compared between three different time-points during Lekhana Basti treatment. Six protein markers overexpressed during obesity were downregulated post Lekhana Basti treatment, while 2 markers increased in abundance post-treatment. CONCLUSION: To our knowledge, this is the first study to isolate and identify urine EV protein abundance profiles from obese female participants of India. The study results indicate significant changes in the differential expression profile of 8 hub proteins involved in obesity, after Lekhana Basti treatment. The biomarker signature of the pilot study indicates the role of Ayurveda treatment and the possible pathways involved in the treatment of Obesity. Further, this study underlines the specificity of urine exosomes/EV as diagnostic markers as well as the potential of Ayurveda treatment in effective management of obesity.