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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

Expression of candidate urine biomarkers of ADR-induced nephropathy during six stages. A) Twelve proteins shared an overall increasing trend in relative abundance. B) Nine proteins shared an overall decreasing trend. C) Two proteins changed at the early stage. The x axis represents different stages; the y axis represents the normalized abundance identified by the LC-MS Progenesis software.
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Figure 3: Expression of candidate urine biomarkers of ADR-induced nephropathy during six stages. A) Twelve proteins shared an overall increasing trend in relative abundance. B) Nine proteins shared an overall decreasing trend. C) Two proteins changed at the early stage. The x axis represents different stages; the y axis represents the normalized abundance identified by the LC-MS Progenesis software.

Mentions: At the discovery stage, eighteen samples of three rats (six time points each) were individually analyzed by UPLC coupled with Triple-TOF 5600 MS. LC-MS Progenesis software was used to calculate the normalized abundance of each protein by measuring the peak area intensity. By label-free quantitative and statistical analyses, 23 proteins met the following criteria: (1) compared with day 0, fold change > 2 in each rat and (2) p value < 0.05 (data were analyzed by repeated measure ANOVA). Among the 23 changed proteins, 20 proteins were annotated as glycoproteins in the UniProt database, 12 proteins shared an overall increasing trend in relative abundance, and 9 proteins shared an overall decreasing trend (Figure 3). The changed proteins are listed in Additional file 1 and the label-free quantitation data are shown in Additional file 2.Three trends were observed in these changed proteins during ADR-induced nephropathy progression. The first was a gradual increase, with examples including afamin and ceruloplasmin. Urinary afamin increased only 1.16-fold on the third day after ADR injection, remained elevated compared with normal urine samples, and peaked at 5.63-fold on day 23 after the ADR injection. The second was a gradual decrease, with examples including cadherin-2 and aggrecan core protein. Cadherin-2 decreased 1.1-fold after 3 days and continued to decrease in the following days. The third, which includes fetuin-B and beta-2-microglobulin, was early changes with distinct patterns. Fetuin-B decreased 2.39-fold on day 3 and remained unchanged during the following days until increasing 1.79-fold on day 23. Beta-2-microglobulin increased 1.7-fold on day 3, decreased 2.83-fold on day 7, and gradually increased 1.69-fold on day 23, compared with the normal samples. The time-dependent changes of 23 representative proteins from three rats are shown in Figure 3.


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)

Expression of candidate urine biomarkers of ADR-induced nephropathy during six stages. A) Twelve proteins shared an overall increasing trend in relative abundance. B) Nine proteins shared an overall decreasing trend. C) Two proteins changed at the early stage. The x axis represents different stages; the y axis represents the normalized abundance identified by the LC-MS Progenesis software.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Expression of candidate urine biomarkers of ADR-induced nephropathy during six stages. A) Twelve proteins shared an overall increasing trend in relative abundance. B) Nine proteins shared an overall decreasing trend. C) Two proteins changed at the early stage. The x axis represents different stages; the y axis represents the normalized abundance identified by the LC-MS Progenesis software.
Mentions: At the discovery stage, eighteen samples of three rats (six time points each) were individually analyzed by UPLC coupled with Triple-TOF 5600 MS. LC-MS Progenesis software was used to calculate the normalized abundance of each protein by measuring the peak area intensity. By label-free quantitative and statistical analyses, 23 proteins met the following criteria: (1) compared with day 0, fold change > 2 in each rat and (2) p value < 0.05 (data were analyzed by repeated measure ANOVA). Among the 23 changed proteins, 20 proteins were annotated as glycoproteins in the UniProt database, 12 proteins shared an overall increasing trend in relative abundance, and 9 proteins shared an overall decreasing trend (Figure 3). The changed proteins are listed in Additional file 1 and the label-free quantitation data are shown in Additional file 2.Three trends were observed in these changed proteins during ADR-induced nephropathy progression. The first was a gradual increase, with examples including afamin and ceruloplasmin. Urinary afamin increased only 1.16-fold on the third day after ADR injection, remained elevated compared with normal urine samples, and peaked at 5.63-fold on day 23 after the ADR injection. The second was a gradual decrease, with examples including cadherin-2 and aggrecan core protein. Cadherin-2 decreased 1.1-fold after 3 days and continued to decrease in the following days. The third, which includes fetuin-B and beta-2-microglobulin, was early changes with distinct patterns. Fetuin-B decreased 2.39-fold on day 3 and remained unchanged during the following days until increasing 1.79-fold on day 23. Beta-2-microglobulin increased 1.7-fold on day 3, decreased 2.83-fold on day 7, and gradually increased 1.69-fold on day 23, compared with the normal samples. The time-dependent changes of 23 representative proteins from three rats are shown in Figure 3.

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