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Calcium oxalate crystals and oxalate induce an epithelial-to-mesenchymal transition in the proximal tubular epithelial cells: Contribution to oxalate kidney injury

View Article: PubMed Central - PubMed

ABSTRACT

TGF-β1 is the main mediator of epithelial-to-mesenchymal transition (EMT). Hyperoxaluria induces crystalluria, interstitial fibrosis, and progressive renal failure. This study analyzed whether hyperoxaluria is associated with TGF-β1 production and kidney fibrosis in mice and if oxalate or calcium oxalate (CaOx) could induce EMT in proximal tubule cells (HK2) and therefore contribute to the fibrotic process. Hyperoxaluria was induced by adding hydroxyproline and ethylene glycol to the mice’s drinking water for up to 60 days. Renal function and oxalate and urinary crystals were evaluated. Kidney collagen production and TGF-β1 expression were assessed. EMT was analyzed in vitro according to TGF-β1 production, phenotypic characterization, invasion, cell migration, gene and protein expression of epithelial and mesenchymal markers. Hyperoxaluric mice showed a decrease in renal function and an increase in CaOx crystals and Ox urinary excretion. The deposition of collagen in the renal interstitium was observed. HK2 cells stimulated with Ox and CaOx exhibited a decreased expression of epithelial as well as increased expression mesenchymal markers; these cells presented mesenchymal phenotypic changes, migration, invasiveness capability and TGF-β1 production, characterizing EMT. Treatment with BMP-7 or its overexpression in HK2 cells was effective at preventing it. This mechanism may contribute to the fibrosis observed in hyperoxaluria.

No MeSH data available.


Related in: MedlinePlus

Hyperoxaluria increased the expression of renal fibrosis markers.The stimulation with ethylene glycol during 60 days (ETG 60 D) and hydroxyproline during 60 days (HPL 60 D) increased the expression of TGF-β1 according to immunoblot image (A) and percentage of immunostaining area (C). There was an increase in the production of collagen type I (yellow to red tone) and collagen type III (greenish tone), showed by picrosirius red staining (E and G). Densitometric quantification of western blot bands (B), immunohistochemistry positive area (D) and collagen staining by picrosirius red (F and H) using ImageJ software. There was a significant increase in TGF-β1 protein expression and collagen production in HPL and ETG treated animals in comparison to controls (CTL) animals. Data are presented as means ± standard errors. N = 5 for each group. (*) Indicates significant differences compared with the control group at p < 0.05.
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f2: Hyperoxaluria increased the expression of renal fibrosis markers.The stimulation with ethylene glycol during 60 days (ETG 60 D) and hydroxyproline during 60 days (HPL 60 D) increased the expression of TGF-β1 according to immunoblot image (A) and percentage of immunostaining area (C). There was an increase in the production of collagen type I (yellow to red tone) and collagen type III (greenish tone), showed by picrosirius red staining (E and G). Densitometric quantification of western blot bands (B), immunohistochemistry positive area (D) and collagen staining by picrosirius red (F and H) using ImageJ software. There was a significant increase in TGF-β1 protein expression and collagen production in HPL and ETG treated animals in comparison to controls (CTL) animals. Data are presented as means ± standard errors. N = 5 for each group. (*) Indicates significant differences compared with the control group at p < 0.05.

Mentions: The expression of TGF-β1 (Fig. 2A and C) and the deposition of collagen fibers (type I yellow to red tone and type III greenish tone) (Fig. 2E and G) are shown in Fig. 2. Hyperoxaluric mice treated with HPL or ETG increased TGF-β1 expression (Fig. 2A–D) and collagen I and III fibers (Fig. 2E–H) in comparison to the control animals.


Calcium oxalate crystals and oxalate induce an epithelial-to-mesenchymal transition in the proximal tubular epithelial cells: Contribution to oxalate kidney injury
Hyperoxaluria increased the expression of renal fibrosis markers.The stimulation with ethylene glycol during 60 days (ETG 60 D) and hydroxyproline during 60 days (HPL 60 D) increased the expression of TGF-β1 according to immunoblot image (A) and percentage of immunostaining area (C). There was an increase in the production of collagen type I (yellow to red tone) and collagen type III (greenish tone), showed by picrosirius red staining (E and G). Densitometric quantification of western blot bands (B), immunohistochemistry positive area (D) and collagen staining by picrosirius red (F and H) using ImageJ software. There was a significant increase in TGF-β1 protein expression and collagen production in HPL and ETG treated animals in comparison to controls (CTL) animals. Data are presented as means ± standard errors. N = 5 for each group. (*) Indicates significant differences compared with the control group at p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5384284&req=5

f2: Hyperoxaluria increased the expression of renal fibrosis markers.The stimulation with ethylene glycol during 60 days (ETG 60 D) and hydroxyproline during 60 days (HPL 60 D) increased the expression of TGF-β1 according to immunoblot image (A) and percentage of immunostaining area (C). There was an increase in the production of collagen type I (yellow to red tone) and collagen type III (greenish tone), showed by picrosirius red staining (E and G). Densitometric quantification of western blot bands (B), immunohistochemistry positive area (D) and collagen staining by picrosirius red (F and H) using ImageJ software. There was a significant increase in TGF-β1 protein expression and collagen production in HPL and ETG treated animals in comparison to controls (CTL) animals. Data are presented as means ± standard errors. N = 5 for each group. (*) Indicates significant differences compared with the control group at p < 0.05.
Mentions: The expression of TGF-β1 (Fig. 2A and C) and the deposition of collagen fibers (type I yellow to red tone and type III greenish tone) (Fig. 2E and G) are shown in Fig. 2. Hyperoxaluric mice treated with HPL or ETG increased TGF-β1 expression (Fig. 2A–D) and collagen I and III fibers (Fig. 2E–H) in comparison to the control animals.

View Article: PubMed Central - PubMed

ABSTRACT

TGF-&beta;1 is the main mediator of epithelial-to-mesenchymal transition (EMT). Hyperoxaluria induces crystalluria, interstitial fibrosis, and progressive renal failure. This study analyzed whether hyperoxaluria is associated with TGF-&beta;1 production and kidney fibrosis in mice and if oxalate or calcium oxalate (CaOx) could induce EMT in proximal tubule cells (HK2) and therefore contribute to the fibrotic process. Hyperoxaluria was induced by adding hydroxyproline and ethylene glycol to the mice&rsquo;s drinking water for up to 60 days. Renal function and oxalate and urinary crystals were evaluated. Kidney collagen production and TGF-&beta;1 expression were assessed. EMT was analyzed in vitro according to TGF-&beta;1 production, phenotypic characterization, invasion, cell migration, gene and protein expression of epithelial and mesenchymal markers. Hyperoxaluric mice showed a decrease in renal function and an increase in CaOx crystals and Ox urinary excretion. The deposition of collagen in the renal interstitium was observed. HK2 cells stimulated with Ox and CaOx exhibited a decreased expression of epithelial as well as increased expression mesenchymal markers; these cells presented mesenchymal phenotypic changes, migration, invasiveness capability and TGF-&beta;1 production, characterizing EMT. Treatment with BMP-7 or its overexpression in HK2 cells was effective at preventing it. This mechanism may contribute to the fibrosis observed in hyperoxaluria.

No MeSH data available.


Related in: MedlinePlus