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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

Semi-quantitative western blot analysis of seven proteins. Semi-quantitation of albumin, serotransferrin, alpha-1-antiproteinase, afamin, ceruloplasmin, plasminogen and AMBP determined by western blot analysis from ten additional rats at six time points each. The two western blots for each protein shown are representative of the group of ten.
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Figure 4: Semi-quantitative western blot analysis of seven proteins. Semi-quantitation of albumin, serotransferrin, alpha-1-antiproteinase, afamin, ceruloplasmin, plasminogen and AMBP determined by western blot analysis from ten additional rats at six time points each. The two western blots for each protein shown are representative of the group of ten.

Mentions: The urine samples from ten more rat models at six time points each were collected for validation. Of 23 changed proteins with disease development, nine altered proteins (albumin, serotransferrin, alpha-1-antiproteinase, afamin, ceruloplasmin, plasminogen, kininogen, alpha-2-HS-glycoprotein and alpha 1 microglobulin [AMBP]) identified in our proteomic study with human orthologs at relatively high abundance that also had commercially available antibodies were selected to be analyzed by semi-quantitative western blot analysis (FigureĀ 4). Kininogen-1 and alpha-2-HS-glycoprotein were not detected by western blot. The changes in the other seven proteins were consistent with the LC-MS/MS results.


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)

Semi-quantitative western blot analysis of seven proteins. Semi-quantitation of albumin, serotransferrin, alpha-1-antiproteinase, afamin, ceruloplasmin, plasminogen and AMBP determined by western blot analysis from ten additional rats at six time points each. The two western blots for each protein shown are representative of the group of ten.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Semi-quantitative western blot analysis of seven proteins. Semi-quantitation of albumin, serotransferrin, alpha-1-antiproteinase, afamin, ceruloplasmin, plasminogen and AMBP determined by western blot analysis from ten additional rats at six time points each. The two western blots for each protein shown are representative of the group of ten.
Mentions: The urine samples from ten more rat models at six time points each were collected for validation. Of 23 changed proteins with disease development, nine altered proteins (albumin, serotransferrin, alpha-1-antiproteinase, afamin, ceruloplasmin, plasminogen, kininogen, alpha-2-HS-glycoprotein and alpha 1 microglobulin [AMBP]) identified in our proteomic study with human orthologs at relatively high abundance that also had commercially available antibodies were selected to be analyzed by semi-quantitative western blot analysis (FigureĀ 4). Kininogen-1 and alpha-2-HS-glycoprotein were not detected by western blot. The changes in the other seven proteins were consistent with the LC-MS/MS results.

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