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Dynamic changes of urinary proteins in a focal segmental glomerulosclerosis rat model.

Zhao M, Li M, Li X, Shao C, Yin J, Gao Y - Proteome Sci (2014)

Bottom Line: To reduce the effects of both genetic and environmental factors on the urinary proteome, this study used a rat model of adriamycin-induced nephropathy resembling human focal segmental glomerulosclerosis (FSGS) development.Of 23 changed proteins with disease development, 20 have human orthologs, and 13 proteins were identified as stable in normal human urine, meaning that changes in these proteins are more likely to reflect disease.Seven proteins were selected for verification in ten more rats as markers closely associated with disease severity by western blot.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathophysiology, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing 100005, China.

ABSTRACT

Background: In contrast to blood, which has mechanisms to maintain a homeostatic internal environment, urine is more likely to reflect changes in the body. As urine accumulates all types of changes, identifying the precise cause of changes in the urine proteome is challenging and crucial in biomarker discovery. To reduce the effects of both genetic and environmental factors on the urinary proteome, this study used a rat model of adriamycin-induced nephropathy resembling human focal segmental glomerulosclerosis (FSGS) development.

Results: Urine samples were collected at before adriamycin administration and day3, 7, 11, 15 and 23 after. Urinary proteins were profiled by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Of 23 changed proteins with disease development, 20 have human orthologs, and 13 proteins were identified as stable in normal human urine, meaning that changes in these proteins are more likely to reflect disease. Fifteen of the identified proteins have not been established to function in FSGS development. Seven proteins were selected for verification in ten more rats as markers closely associated with disease severity by western blot.

Conclusion: We identified proteins changed in different stages of FSGS in rat models, which may aid in biomarker development and the understanding of FSGS pathogenesis.

No MeSH data available.


Related in: MedlinePlus

Workflow of protein identification in the FSGS rat model. Urine was collected at days 0, 3, 7, 11, 15 and 23 after ADR administration, and urinary proteins were enriched by ConA agarose followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) identification. Several proteins were verified in ten more rats by western blot analysis.
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Figure 1: Workflow of protein identification in the FSGS rat model. Urine was collected at days 0, 3, 7, 11, 15 and 23 after ADR administration, and urinary proteins were enriched by ConA agarose followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) identification. Several proteins were verified in ten more rats by western blot analysis.

Mentions: In the late phase of FSGS, massive irreversible proteinuria commonly occurs after podocyte injury, which increases the difficulty of identifying some low-abundance proteins by MS. Concanavalin A (ConA) enrichment is frequently used to enrich for N-linked glycoproteins in both plasma and urine proteomic studies [10,11]. As many existing biomarkers are glycoproteins, in this study, ConA-enriched urinary protein was evaluated to monitor the dynamic changes during FSGS development (FigureĀ 1).


Dynamic changes of urinary proteins in a focal segmental glomerulosclerosis rat model.

Zhao M, Li M, Li X, Shao C, Yin J, Gao Y - Proteome Sci (2014)

Workflow of protein identification in the FSGS rat model. Urine was collected at days 0, 3, 7, 11, 15 and 23 after ADR administration, and urinary proteins were enriched by ConA agarose followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) identification. Several proteins were verified in ten more rats by western blot analysis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4109389&req=5

Figure 1: Workflow of protein identification in the FSGS rat model. Urine was collected at days 0, 3, 7, 11, 15 and 23 after ADR administration, and urinary proteins were enriched by ConA agarose followed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) identification. Several proteins were verified in ten more rats by western blot analysis.
Mentions: In the late phase of FSGS, massive irreversible proteinuria commonly occurs after podocyte injury, which increases the difficulty of identifying some low-abundance proteins by MS. Concanavalin A (ConA) enrichment is frequently used to enrich for N-linked glycoproteins in both plasma and urine proteomic studies [10,11]. As many existing biomarkers are glycoproteins, in this study, ConA-enriched urinary protein was evaluated to monitor the dynamic changes during FSGS development (FigureĀ 1).

Bottom Line: To reduce the effects of both genetic and environmental factors on the urinary proteome, this study used a rat model of adriamycin-induced nephropathy resembling human focal segmental glomerulosclerosis (FSGS) development.Of 23 changed proteins with disease development, 20 have human orthologs, and 13 proteins were identified as stable in normal human urine, meaning that changes in these proteins are more likely to reflect disease.Seven proteins were selected for verification in ten more rats as markers closely associated with disease severity by western blot.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathophysiology, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing 100005, China.

ABSTRACT

Background: In contrast to blood, which has mechanisms to maintain a homeostatic internal environment, urine is more likely to reflect changes in the body. As urine accumulates all types of changes, identifying the precise cause of changes in the urine proteome is challenging and crucial in biomarker discovery. To reduce the effects of both genetic and environmental factors on the urinary proteome, this study used a rat model of adriamycin-induced nephropathy resembling human focal segmental glomerulosclerosis (FSGS) development.

Results: Urine samples were collected at before adriamycin administration and day3, 7, 11, 15 and 23 after. Urinary proteins were profiled by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Of 23 changed proteins with disease development, 20 have human orthologs, and 13 proteins were identified as stable in normal human urine, meaning that changes in these proteins are more likely to reflect disease. Fifteen of the identified proteins have not been established to function in FSGS development. Seven proteins were selected for verification in ten more rats as markers closely associated with disease severity by western blot.

Conclusion: We identified proteins changed in different stages of FSGS in rat models, which may aid in biomarker development and the understanding of FSGS pathogenesis.

No MeSH data available.


Related in: MedlinePlus