Table 1.
Primers used for real-time PCR.
Fig 1.
Hemi-nephrectomized rats fed a high-P diet.
(a) Serum Cr concentration. (b) The 24-hr CCr was calculated using the following formula: (urine Cr concentration×24-hour urine volume)/(serum Cr concentration×1440×body weight). (c) Serum P concentration. (d) Serum Ca concentration. (e) The 24-hr U-P was calculated using the following formula: (urine P concentration x 24-hour urine volume)/body weight. (f) The 24-hr U-Ca was calculated using the following formula: (urine Ca concentration×24-hour urine volume)/body weight. (g) Serum 1,25(OH)2D3 concentration. (h) Serum intact PTH concentration. (i) αKlotho mRNA expression in the kidney. (j) NaPi-IIa mRNA expression in the kidney. (k) NaPi-IIc mRNA expression in the kidney. (l) CYP27B1 mRNA expression in the kidney. (m) TGFβ1 mRNA expression in the kidney. (n) Col1a1 mRNA expression in the kidney. (o) Osteopontin mRNA expression in the kidney. (p) Osteocalcin mRNA expression in the kidney. Each value represents the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001, NP sham group vs HP sham group or NP Nx group vs HP Nx group; NP sham group (n = 8), NP Nx group (n = 7), HP sham group (n = 7) and HP Nx group (n = 9). (i-p) NP sham group was used as a normalization control.
Fig 2.
FGF23 expression in hemi-nephrectomized rats fed a high-P diet.
(a) Serum FGF23 concentration. (b) FGF23 mRNA expression in the bone. NP sham group was used as a normalization control. (c) FGF23 mRNA expression in the kidney. NP sham group was used as a normalization control. (d) Western blot of FGF23 in the kidney. GAPDH was used as an internal control. Each value shown represents the mean ± SEM; *P<0.05, **P<0.01; NP sham group (n = 8), NP Nx group (n = 7), HP sham group (n = 7) and HP Nx group (n = 9).
Fig 3.
Histology on the kidney in hemi-nephrectomized rats fed a high-P diet.
HE: hematoxylin-eosin staining. MT: Masson’s trichrome staining as an evaluation of fibrosis. VK: Von Kossa staining as an evaluation of calcification. ×200: high magnification.
Fig 4.
Immunohistochemistry and in situ hybridization in the kidney of hemi-nephrectomized rats fed a high-P diet.
FGF23: FGF23 immunohistochemistry (brown). Osteopontin: osteopontin immunohistochemistry (brown). ×200: high magnification. FGF23 in situ hybridization (red spots. arrow: positive cells). ×400: high magnification.
Fig 5.
Immunofluorescence in the kidney of hemi-nephrectomized rats fed a high-P diet.
FGF23 immunofluorescence (green). αSMA immunofluorescence (red). Merge: FGF23 (green), αSMA (red), and DAPI (blue). ×400: high magnification.
Fig 6.
FGF23 mRNA expression in HK2 cells.
(a) FGF23 mRNA expression in HK2 cells stimulated with TGFβ1. (b) Osteopontin mRNA expression in HK2 cells stimulated with TGFβ1. (c) E-cadherin mRNA expression in HK2 cells stimulated with TGFβ1. (d) PAI1 mRNA expression in HK2 cells stimulated with TGFβ1. (m) FGF23 mRNA expression in HK2 cells stimulated with 1,25(OH)2D3. (l) FGF23 mRNA expression in HK2 cells stimulated with high-P levels. Each value represents the mean ± SEM; *P<0.05, **P<0.01, ***P<0.001, compared with the vehicle (each group: n = 3). Vehicle group was used as a normalization control.
Fig 7.
Partial nephrectomy rat model.
(a) Change in serum Cr concentration. (b) The 24-hr CCr. (c) Serum P concentration. (d) Serum Ca concentration. (e) The 24-hr U-P. (f) The 24-hr U-Ca. (g) Serum 1,25(OH)2D3 concentration. (h) Serum intact PTH concentration. (i) αKlotho mRNA expression in the kidney. (j) NaPi-IIa mRNA expression in the kidney. (k) NaPi-IIc mRNA expression in the kidney. (l) CYP27B1 mRNA expression in the kidney. (m) TGFβ1 mRNA expression in the kidney. (n) Col1a1 mRNA expression in the kidney. (o) Osteopontin mRNA expression in the kidney. (p) Osteocalcin mRNA expression in the kidney. Each value represents the mean ± SEM; *P<0.05, **P<0.01, ***P<0.001, sham group vs PN mild group or sham group vs PN severe group. Sham group (n = 6), PN mild group (n = 6), PN severe group (n = 6). (i-p) Sham group was used as a normalization control.
Fig 8.
FGF23 expression in the partial nephrectomy rat model.
(a) Serum FGF23 concentration. (b) FGF23 mRNA expression in the kidney. Sham group was used as a normalization control. (c) Western blot of FGF23 in the kidney. (d) Histology in the kidney. HE: hematoxylin-eosin staining. MT: Masson’s trichrome staining to evaluate fibrosis. VK: Von Kossa staining to evaluate calcification. *P<0.05, **P<0.01, ***P<0.001; sham group (n = 6), partial nephrectomy mild group (PN mild) (n = 6), partial nephrectomy severe group (PN severe) (n = 6).
Fig 9.
Immunohistochemistry and in situ hybridization in the kidney of partial nephrectomy rat model.
FGF23: FGF23 immunohistochemistry (brown). Osteopontin: osteopontin immunohistochemistry (brown). ×200: high magnification. FGF23 in situ hybridization (red spots; arrow: positive cells). ×400: high magnification.
Fig 10.
Rat model of DXR-induced renal failure.
(a) Serum Cr concentration. (b) Serum P concentration. (c) Serum Ca concentration. (d) αKlotho mRNA expression in the kidney. (e) NaPi-IIa mRNA expression in the kidney. (f) NaPi-IIc mRNA expression in the kidney. (g) CYP27B1 mRNA expression in the kidney. (h) FGF23 mRNA expression in the kidney. (j) TGFβ1 mRNA expression in the kidney. (j) Col1a1 mRNA expression in the kidney. (k) Osteopontin mRNA expression in the kidney. (l) Osteocalcin mRNA expression in the kidney. (m) Histology in the kidney. MT: Masson’s trichrome staining to evaluate fibrosis. Each value represents the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001,; vehicle group (n = 5) and DXR group (n = 5). (d-l) Vehicle group was used as a normalization control.