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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

UCH-L1 overexpression induced podocyte hypermotility as assessed by cell migration assay. (A) Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector. Control cells were treated with vehicle vector or were not treated. Then subsequently scratch was processed using a 0.1 mL pipette. The observation was made immediately after scratch (0:00 h) and at 24:00 h, Original magnification, 200×; (B) quantification of the cell migration area by computerized morphometric analysis. Data are given as means + SD. n = 20 areas from three independent experiments. * p < 0.05 versus control group and control infection group, respectively.
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ijms-17-01404-f006: UCH-L1 overexpression induced podocyte hypermotility as assessed by cell migration assay. (A) Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector. Control cells were treated with vehicle vector or were not treated. Then subsequently scratch was processed using a 0.1 mL pipette. The observation was made immediately after scratch (0:00 h) and at 24:00 h, Original magnification, 200×; (B) quantification of the cell migration area by computerized morphometric analysis. Data are given as means + SD. n = 20 areas from three independent experiments. * p < 0.05 versus control group and control infection group, respectively.

Mentions: Further, a cell migration assay was carried to assess the motility of podocytes in UCH-L1 adenoviral vector-infected podocytes and control groups. As shown in Figure 6, compared with both control groups, UCH-L1 infection could significantly prompt the motility of podocytes, lessen the distances between the leading edges of the migrating podocytes. The results suggested that UCH-L1 overexpression may empower hypermotility of podocytes. Combining the above results, it was clear that UCH-L1 overexpression could affect the morphological and functional changes of podocytes.


Wnt/ β -Catenin Signaling Mediated-UCH-L1 Expression in Podocytes of Diabetic Nephropathy
UCH-L1 overexpression induced podocyte hypermotility as assessed by cell migration assay. (A) Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector. Control cells were treated with vehicle vector or were not treated. Then subsequently scratch was processed using a 0.1 mL pipette. The observation was made immediately after scratch (0:00 h) and at 24:00 h, Original magnification, 200×; (B) quantification of the cell migration area by computerized morphometric analysis. Data are given as means + SD. n = 20 areas from three independent experiments. * p < 0.05 versus control group and control infection group, respectively.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-17-01404-f006: UCH-L1 overexpression induced podocyte hypermotility as assessed by cell migration assay. (A) Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector. Control cells were treated with vehicle vector or were not treated. Then subsequently scratch was processed using a 0.1 mL pipette. The observation was made immediately after scratch (0:00 h) and at 24:00 h, Original magnification, 200×; (B) quantification of the cell migration area by computerized morphometric analysis. Data are given as means + SD. n = 20 areas from three independent experiments. * p < 0.05 versus control group and control infection group, respectively.
Mentions: Further, a cell migration assay was carried to assess the motility of podocytes in UCH-L1 adenoviral vector-infected podocytes and control groups. As shown in Figure 6, compared with both control groups, UCH-L1 infection could significantly prompt the motility of podocytes, lessen the distances between the leading edges of the migrating podocytes. The results suggested that UCH-L1 overexpression may empower hypermotility of podocytes. Combining the above results, it was clear that UCH-L1 overexpression could affect the morphological and functional changes of podocytes.

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/&beta;-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 &beta;-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/&beta;-catenin/UCH-L1 may be a new potential therapy method in the treatment of DN in future.

No MeSH data available.


Related in: MedlinePlus