In depth mass spectrometry based proteomic approach to explore the effect of curcumin in IL17A mediated alveolar barrier injury

Abstract

Introduction: Lung barrier function is collaboratively formed by tight junction proteins (TJP) contributing in overdrive activation of alveolar EMT. The critical role of inflammatory cytokine mediated lung epithelial barrier in the pathogenesis and recovery from acute lung injury remains unknown. Here, we describe new link between IL-17A and TJP with p53-fibrinolytic system via studying the effect ofl curcumin during compromised lung barrier integrity. Proteomic profiling confirmed the activityof barrier proteins triggering p53-fibrinolytic and ubiquitin proteins via drawing an intervention pathway in restoration of these barrier proteins.

Method: Basal alveolar epithelial cells and C57BL/6 mice were used. BLM and IL-17A were used to exert alveolar injury followed by curcumin intervention. Proteomic data set revealed novel claudins such as claudin-16, claudin-19, claudin-20 and claudin-25 with other EMT proteins validated by immuno blot, qRT-PCR and pathological staining. IL-17A, p53 and PAI-1 were knocked down to study the activity of barrier proteins. Proteomic profiling on clinical samples confirmed findings.

Results: Proteomic profiling revealed decrease levels of TJPs and claudins which were restored by curcumin. Increase levels of EMT proteins and fibrinolytic proteins was inhibited by curcumin trigeering ubiquitin pathways. Our analysis provides insight into mechanisms regulating ubiquitination. Knocking down important genes confirm role of TJPs in p53-fibrinolytic system. Clinical proteomics revealed novel biomarkers in lungs.

Method: This study reveals curcumin heals lung barrier injury revealing novel proteins in detection of lung fibrosis.