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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

Dynamic changes in ADR-treated rats. A) Urinary protein to creatinine ratios in six phases of ADR treated rats. The data are expressed as the mean ± standard deviation (n = 13, *p < 0.05 for Experiment vs. Control). The paired t test was used to assess the significance of the differences between groups. B) Histological changes in ADR-treated rats (HE staining, 400×). Compared with the normal condition, on day 23, glomerular lesions with mesangial matrix expansion could be detected; on day 50, glomerular sclerosis and severe interstitial expansion were found in many glomeruli. C) Urinary protein and ConA-enriched urinary protein pattern of one FSGS model rat. Lane 1: Marker. Lanes 2–7, urinary proteins and ConA-enriched urinary proteins at days 0, 3, 7, 11, 15 and 23 respectively.
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Figure 2: Dynamic changes in ADR-treated rats. A) Urinary protein to creatinine ratios in six phases of ADR treated rats. The data are expressed as the mean ± standard deviation (n = 13, *p < 0.05 for Experiment vs. Control). The paired t test was used to assess the significance of the differences between groups. B) Histological changes in ADR-treated rats (HE staining, 400×). Compared with the normal condition, on day 23, glomerular lesions with mesangial matrix expansion could be detected; on day 50, glomerular sclerosis and severe interstitial expansion were found in many glomeruli. C) Urinary protein and ConA-enriched urinary protein pattern of one FSGS model rat. Lane 1: Marker. Lanes 2–7, urinary proteins and ConA-enriched urinary proteins at days 0, 3, 7, 11, 15 and 23 respectively.

Mentions: To monitor renal function, the urinary protein to creatinine ratios were measured to determine the proteinuria levels. As shown in Figure 2A, the ratios increased. From day 3 to day 7, the increase was moderate. Marked increases occurred 11 days after ADR induction. To confirm the successful induction of FSGS, the histological characteristics of the rat kidneys on days 23 and 50 were examined. On day 23, the glomerular endothelium and podocyte were partly effaced, foam cells were generated, and fewer glomerular lesions with mesangial matrix expansion were present. On day 50, severe segmental and global sclerosis, mesangial matrix expansion and interstitial fibrosis were present (Figure 2B). All the features indicated that the disease was in the late stage.The urinary proteins at days 0, 3, 7, 11, 15 and 23 were analyzed by SDS-PAGE. Compared with day 0, the expression of the 70-kD protein was gradually increased, and this protein became the most intense band at day 23. The ConA-enriched samples at each stage were also shown in Figure 2C.


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)

Dynamic changes in ADR-treated rats. A) Urinary protein to creatinine ratios in six phases of ADR treated rats. The data are expressed as the mean ± standard deviation (n = 13, *p < 0.05 for Experiment vs. Control). The paired t test was used to assess the significance of the differences between groups. B) Histological changes in ADR-treated rats (HE staining, 400×). Compared with the normal condition, on day 23, glomerular lesions with mesangial matrix expansion could be detected; on day 50, glomerular sclerosis and severe interstitial expansion were found in many glomeruli. C) Urinary protein and ConA-enriched urinary protein pattern of one FSGS model rat. Lane 1: Marker. Lanes 2–7, urinary proteins and ConA-enriched urinary proteins at days 0, 3, 7, 11, 15 and 23 respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Dynamic changes in ADR-treated rats. A) Urinary protein to creatinine ratios in six phases of ADR treated rats. The data are expressed as the mean ± standard deviation (n = 13, *p < 0.05 for Experiment vs. Control). The paired t test was used to assess the significance of the differences between groups. B) Histological changes in ADR-treated rats (HE staining, 400×). Compared with the normal condition, on day 23, glomerular lesions with mesangial matrix expansion could be detected; on day 50, glomerular sclerosis and severe interstitial expansion were found in many glomeruli. C) Urinary protein and ConA-enriched urinary protein pattern of one FSGS model rat. Lane 1: Marker. Lanes 2–7, urinary proteins and ConA-enriched urinary proteins at days 0, 3, 7, 11, 15 and 23 respectively.
Mentions: To monitor renal function, the urinary protein to creatinine ratios were measured to determine the proteinuria levels. As shown in Figure 2A, the ratios increased. From day 3 to day 7, the increase was moderate. Marked increases occurred 11 days after ADR induction. To confirm the successful induction of FSGS, the histological characteristics of the rat kidneys on days 23 and 50 were examined. On day 23, the glomerular endothelium and podocyte were partly effaced, foam cells were generated, and fewer glomerular lesions with mesangial matrix expansion were present. On day 50, severe segmental and global sclerosis, mesangial matrix expansion and interstitial fibrosis were present (Figure 2B). All the features indicated that the disease was in the late stage.The urinary proteins at days 0, 3, 7, 11, 15 and 23 were analyzed by SDS-PAGE. Compared with day 0, the expression of the 70-kD protein was gradually increased, and this protein became the most intense band at day 23. The ConA-enriched samples at each stage were also shown in Figure 2C.

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