Dataset Information

Modulation of proteomic and inflammatory signals by Bradykinin in podocytes.

ABSTRACT: Podocyte damage is one of the hallmarks of diabetic nephropathy leading to proteinuria and kidney damage. The underlying mechanisms of podocyte injury are not well defined. Bradykinin (BK) was shown to contribute to diabetic kidney disease. Here, we evaluated the temporal changes in proteome profile and inflammatory signals of podocytes in response to BK (10-7M). Protein profile was evaluated by liquid chromatography mass Spectrometry (LC-MS/MS) analysis. Proteome profile analysis of podocytes treated with BK (10-7M) for 3 and 6 h, revealed 61 proteins that were differentially altered compared to unstimulated control podocytes. Pathway enrichment analysis suggested inhibition of cell death pathways, engagement of cytoskeletal elements and activation of inflammatory pathways. One of the inflammatory proteins that was identified to be induced by BK treatment is Prostaglandin (PG) H Synthase-2 (Cyclooxygenase-2, COX-2). In addition, BK significantly induced the production and release of PGE2 and this effect was inhibited by both COX-2 and MEK Kinase inhibitors, demonstrating that the production of PGE2 by BK is mediated via COX-2 and MAPK-dependent mechanisms. These findings provide a global understanding of the effector modulated proteome in response to BK and also reveal BK as an important modulator of inflammation and a potential player in podocyte injury.

SUBMITTER: Saoud R

PROVIDER: S-EPMC7270529 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Modulation of proteomic and inflammatory signals by Bradykinin in podocytes.

Saoud Richard R Jaffa Miran A MA Habib Aida A Zhao Jingfu J Al Hariri Moustafa M Zhu Rui R Hasan Anwarul A Ziyadeh Fuad N FN Kobeissy Firas F Mechref Yehia Y Jaffa Ayad A AA

Journal of advanced research 20200528

Podocyte damage is one of the hallmarks of diabetic nephropathy leading to proteinuria and kidney damage. The underlying mechanisms of podocyte injury are not well defined. Bradykinin (BK) was shown to contribute to diabetic kidney disease. Here, we evaluated the temporal changes in proteome profile and inflammatory signals of podocytes in response to BK (10<sup>-7</sup>M). Protein profile was evaluated by liquid chromatography mass Spectrometry (LC-MS/MS) analysis. Proteome profile analysis of ...[more]

PMID: 32518694

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Project description:Diabetic kidney disease is the major cause of end-stage kidney failure worldwide. Podocyte damage is one of the hallmarks of diabetic nephropathy, where the loss of the podocyte filtration barrier leads to proteinuria and progressive renal damage. Bradykinin (BK) is a vasoactive peptide that was shown to play an important role in the progression of renal damage. In this study, we aim to evaluate the changes in protein expression profiles of rat podocytes upon the treatment with BK by an advanced proteomics platform in conjunction with bioinformatics and pathway enrichment analysis. This analysis will help in discovering core effectors whose upstream and downstream protein profile can elucidate signaling mechanisms underlying BK’s modulation of the podocyte biology. Cultured immortalized rat podocytes were treated with BK at 10-7 M for 3 and 6 hours. Proteome profile was evaluated by LC-MS/MS analysis, and pathway enrichment analysis was performed on the resulting differential proteomic data. Proteomic analysis of podocytes treated by BK revealed 61 proteins that were differentially altered (up or downregulated) in response to BK treatment compared to unstimulated control podocytes. Pathway enrichment analysis suggested an inhibition of the cell death pathway and the involvement of elements of the cytoskeletal activity as well as activation of the inflammatory pathways. Of interest, one of the inflammatory proteins that were identified to be induced by BK treatment is Prostaglandin H Synthase-2 (PTGS-2, also known as COX-2). The upregulation of COX-2 by BK treatment was validated by western blot analysis. In addition, treatment of podocytes with BK significantly induced the production and release of PGE2 and this effect was inhibited by both COX-2 and MAPK inhibitors, demonstrating that the production of PGE2 by BK is mediated via a COX-2 and MAPK-dependent mechanisms. The findings of the present study provide a global understanding of the effector modulated proteome in response to BK treatment, and also reveal BK as an important modulator in the arachidonic acid metabolism pathway and a potential player in the progression of diabetic nephropathy.

Project description:Microglia, the resident phagocytes of the central nervous system and one of the key modulators of the innate immune system, have been shown to play a major role in brain insults. Upon activation in response to neuroinflammation, microglia promote the release of inflammatory mediators as well as promote phagocytosis. Plasma prekallikrein (PKall) has been recently implicated as a mediator of neuroinflammation; nevertheless, its role in mediating microglial activation has not been investigated yet. In the current study, we evaluate the mechanisms through which PKall contributes to microglial activation and release of inflammatory cytokines assessing PKall-related receptors and their dynamics. Murine N9-microglial cells were exposed to PKall (2.5 ng/ml), lipopolysaccharide (100 ng/ml), bradykinin (BK, 0.1 μM), and neuronal cell debris (16.5 μg protein/ml). Gene expression of bradykinin 2 receptor (B2KR), protease-activated receptor 2 (PAR-2), along with cytokines and fibrotic mediators were studied. Bioinformatic analysis was conducted to correlate altered protein changes with microglial activation. To assess receptor dynamics, HOE-140 (1 μM) and GB-83 (2 μM) were used to antagonize the B2KR and PAR-2 receptors, respectively. Also, the role of autophagy in modulating microglial response was evaluated. Data from our work indicate that PKall, LPS, BK, and neuronal cell debris resulted in the activation of microglia and enhanced expression/secretion of inflammatory mediators. Elevated increase in inflammatory mediators was attenuated in the presence of HOE-140 and GB-83, implicating the engagement of these receptors in the activation process coupled with an increase in the expression of B2KR and PAR-2. Finally, the inhibition of autophagy significantly enhanced the release of the cytokine IL-6 which were validated via bioinformatics analysis demonstrating the role of PKall in systematic and brain inflammatory processes. Taken together, we demonstrated that PKall can modulate microglial activation via the engagement of PAR-2 and B2KR where PKall acts as a neuromodulator of inflammatory processes.

Dataset Information

Modulation of proteomic and inflammatory signals by Bradykinin in podocytes.

Publications

Modulation of proteomic and inflammatory signals by Bradykinin in podocytes.

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