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Wnt/ β -Catenin Signaling Mediated-UCH-L1 Expression in Podocytes of Diabetic Nephropathy

View Article: PubMed Central - PubMed

ABSTRACT

Increasing studies identified podocyte injury as a key early risk factor resulting in diabetic nephropathy (DN). The ubiquitin carboxy-terminal hydrolase 1 (UCH-L1) participates in podocyte differentiation and injury, which is elevated in the podocytes of a variety of nephritis. Whether UCH-L1 expression is positively related to podocyte injury of DN remains unclear. In this study, elevated expression of UCH-L1 and its intrinsic mechanism in high glucose (HG)-stimulated murine podocytes were investigated using western blot and real-time quantitative PCR. Kidney biopsies of DN patients and health individuals were stained by immunofluorescence (IF) method. The morphological and functional changes of podocytes were tested by F-actin staining and cell migration assay. Results demonstrated that HG induced upregulation of UCH-L1 and activation of the Wnt/β-catenin signaling pathway in podocytes. However, blocking of the Wnt pathway by dickkopf related protein 1 (DKK1) eliminated the above changes. Furthermore, IF staining confirmed that, compared with healthy individuals, the expression of UCH-L1 and β-catenin were obviously increased in kidney biopsy of DN patients. Overexpression of UCH-L1 remodeled its actin cytoskeleton, increased its cell migration and impacted its important proteins. All the findings manifested that Wnt/β-catenin/UCH-L1 may be a new potential therapy method in the treatment of DN in future.

No MeSH data available.


Related in: MedlinePlus

(A) UCH-L1 expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy control persons and DN patients. Synaptopodin showed in red, UCH-L1 showed in green and DAPI showed in blue (n = 6). Original magnification, 200×; (B) β-catenin expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy controls persons and DN patients. Synaptopodin showed in red, β-catenin showed in green and DAPI showed in blue (n = 6). Original magnification, 200×.
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ijms-17-01404-f004: (A) UCH-L1 expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy control persons and DN patients. Synaptopodin showed in red, UCH-L1 showed in green and DAPI showed in blue (n = 6). Original magnification, 200×; (B) β-catenin expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy controls persons and DN patients. Synaptopodin showed in red, β-catenin showed in green and DAPI showed in blue (n = 6). Original magnification, 200×.

Mentions: To investigate the potential role of Wnt/β-catenin/UCH-L1 signaling in podocyte injury in vivo, immunofluorescence staining was used to explore the expression of UCH-L1 and β-catenin in podocytes of diabetic nephropathy patients. We selected typical DN slides to further assess. The typical images of DN were that parts of glomeruli experienced nodular sclerosis, together with increase of mesangial matrix, appearance of fibrin cap and thickening of capillary wall. The healthy control slides had none of the above features. As shown in Figure 4A, double immunofluorescence staining for both synaptopodin (red) and UCH-L1 (green) revealed that UCH-L1 can be evidently positioned in the podocytes (marked by synaptopodin) of diabetic nephropathy patients. Positive staining of UCH-L1 was mainly located at the peripheral area of the capillary tufts in accord with the distribution of synaptopodin. However, in the healthy control slides, there was no obvious positive staining of UCH-L1. As for β-catenin, which was shown in Figure 4B, weak staining was detectable in the healthy control slides, however, strong positive staining was detectable in synaptopodin-positive staining podocytes in diabetic nephropathy patients. These results suggested that in podocytes of diabetic nephropathy, both UCH-L1 and β-catenin were increased, which exhibited consistent results with those of cells experiments in vivo.


Wnt/ β -Catenin Signaling Mediated-UCH-L1 Expression in Podocytes of Diabetic Nephropathy
(A) UCH-L1 expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy control persons and DN patients. Synaptopodin showed in red, UCH-L1 showed in green and DAPI showed in blue (n = 6). Original magnification, 200×; (B) β-catenin expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy controls persons and DN patients. Synaptopodin showed in red, β-catenin showed in green and DAPI showed in blue (n = 6). Original magnification, 200×.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5037684&req=5

ijms-17-01404-f004: (A) UCH-L1 expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy control persons and DN patients. Synaptopodin showed in red, UCH-L1 showed in green and DAPI showed in blue (n = 6). Original magnification, 200×; (B) β-catenin expression in podocytes of diabetic nephropathy patients. Immunofluorescence staining was performed in 4-μm-thick frozen sections from healthy controls persons and DN patients. Synaptopodin showed in red, β-catenin showed in green and DAPI showed in blue (n = 6). Original magnification, 200×.
Mentions: To investigate the potential role of Wnt/β-catenin/UCH-L1 signaling in podocyte injury in vivo, immunofluorescence staining was used to explore the expression of UCH-L1 and β-catenin in podocytes of diabetic nephropathy patients. We selected typical DN slides to further assess. The typical images of DN were that parts of glomeruli experienced nodular sclerosis, together with increase of mesangial matrix, appearance of fibrin cap and thickening of capillary wall. The healthy control slides had none of the above features. As shown in Figure 4A, double immunofluorescence staining for both synaptopodin (red) and UCH-L1 (green) revealed that UCH-L1 can be evidently positioned in the podocytes (marked by synaptopodin) of diabetic nephropathy patients. Positive staining of UCH-L1 was mainly located at the peripheral area of the capillary tufts in accord with the distribution of synaptopodin. However, in the healthy control slides, there was no obvious positive staining of UCH-L1. As for β-catenin, which was shown in Figure 4B, weak staining was detectable in the healthy control slides, however, strong positive staining was detectable in synaptopodin-positive staining podocytes in diabetic nephropathy patients. These results suggested that in podocytes of diabetic nephropathy, both UCH-L1 and β-catenin were increased, which exhibited consistent results with those of cells experiments in vivo.

View Article: PubMed Central - PubMed

ABSTRACT

Increasing studies identified podocyte injury as a key early risk factor resulting in diabetic nephropathy (DN). The ubiquitin carboxy-terminal hydrolase 1 (UCH-L1) participates in podocyte differentiation and injury, which is elevated in the podocytes of a variety of nephritis. Whether UCH-L1 expression is positively related to podocyte injury of DN remains unclear. In this study, elevated expression of UCH-L1 and its intrinsic mechanism in high glucose (HG)-stimulated murine podocytes were investigated using western blot and real-time quantitative PCR. Kidney biopsies of DN patients and health individuals were stained by immunofluorescence (IF) method. The morphological and functional changes of podocytes were tested by F-actin staining and cell migration assay. Results demonstrated that HG induced upregulation of UCH-L1 and activation of the Wnt/β-catenin signaling pathway in podocytes. However, blocking of the Wnt pathway by dickkopf related protein 1 (DKK1) eliminated the above changes. Furthermore, IF staining confirmed that, compared with healthy individuals, the expression of UCH-L1 and β-catenin were obviously increased in kidney biopsy of DN patients. Overexpression of UCH-L1 remodeled its actin cytoskeleton, increased its cell migration and impacted its important proteins. All the findings manifested that Wnt/β-catenin/UCH-L1 may be a new potential therapy method in the treatment of DN in future.

No MeSH data available.


Related in: MedlinePlus