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  • Author: Trairak Pisitkun x
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Panthip Rattanasinganchan, Kittipat Sopitthummakhun, Kent Doi, Xuzhen Hu, D. Michael Payne, Trairak Pisitkun and Asada Leelahavanichkul



Chronic tubulointerstitial (TI) fibrosis is difficult to diagnose. Urinary exosomes contain many potential biomarkers, and may be useful to identify biomarkers of TI fibrosis. However, a suitable animal model to investigate urinary exosome biomarkers for TI fibrosis has not yet been specifically identified.


To test the suitability of a folic-acid induced rat model of renal injury for identifying potential biomarkers of fibrosis from urinary exosomes.


We employed a rat model of renal injury using a single intraperitoneal injection of folic acid. Urinary exosomes were isolated and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS)


We demonstrated severe acute kidney injury after folic acid injection by a 6-fold decrease in creatinine clearance at day 2, severe inflammatory cell infiltration in the TI area at day 7, and renal fibrosis at day 14. We collected 24 h urine samples from folic acid-injected rats and vehicle-injected control rats at days 7 and 14, then isolated urinary exosomes using a differential centrifugation method. We found that levels of exosomal marker proteins ALG-2-interacting protein 1 (ALIX; PDCD6IP) and tumor susceptibility gene 101 (TSG101) were higher in folic acid-injected rats than in control rats. LC-MS/MS analysis of urinary exosomal proteins from the folic acid-injected and control rats identified 372 proteins, including various potential TI fibrosis biomarkers


We demonstrated the feasibility of using a folic acid-induced rat model of TI fibrosis for urinary proteomic analysis. These rats excreted sufficient urinary exosomes permitting the large-scale analysis and discovery of potential biomarkers of renal fibrosis.