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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 changes of expression of several proteins in podocytes. Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector for 48 h. Control cells were treated with vehicle vector or were not treated for 48 h. Podocytes lysate proteins (60 μg) were analyzed respectively by western blot using murine corresponding antibodies and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1, synaptopodin, CD2AP, nephrin and Snail; (B) corresponding statistic histogram of UCH-L1, synaptopodin, CD2AP, nephrin, and Snail protein expression. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 both versus control group and control infection group.
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ijms-17-01404-f007: UCH-L1 overexpression induced changes of expression of several proteins in podocytes. Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector for 48 h. Control cells were treated with vehicle vector or were not treated for 48 h. Podocytes lysate proteins (60 μg) were analyzed respectively by western blot using murine corresponding antibodies and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1, synaptopodin, CD2AP, nephrin and Snail; (B) corresponding statistic histogram of UCH-L1, synaptopodin, CD2AP, nephrin, and Snail protein expression. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 both versus control group and control infection group.

Mentions: To further investigate the effect of increased UCH-L1 in podocytes, several important molecules of podocytes were assessed after differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector for 48 h. As shown in Figure 7, expression of UCH-L1 in UCH-L1 infected-group was significantly high than that of control group and control infection group, which demonstrated the successful infection. The successful infection decreased the expression of synaptopodin, CD2AP and nephrin and increased that of Snail. These results demonstrated that UCH-L1 could change the expression of important proteins in podocytes, which may correspondingly affect its structure and function.


Wnt/ β -Catenin Signaling Mediated-UCH-L1 Expression in Podocytes of Diabetic Nephropathy
UCH-L1 overexpression induced changes of expression of several proteins in podocytes. Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector for 48 h. Control cells were treated with vehicle vector or were not treated for 48 h. Podocytes lysate proteins (60 μg) were analyzed respectively by western blot using murine corresponding antibodies and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1, synaptopodin, CD2AP, nephrin and Snail; (B) corresponding statistic histogram of UCH-L1, synaptopodin, CD2AP, nephrin, and Snail protein expression. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 both versus control group and control infection group.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-17-01404-f007: UCH-L1 overexpression induced changes of expression of several proteins in podocytes. Differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector for 48 h. Control cells were treated with vehicle vector or were not treated for 48 h. Podocytes lysate proteins (60 μg) were analyzed respectively by western blot using murine corresponding antibodies and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1, synaptopodin, CD2AP, nephrin and Snail; (B) corresponding statistic histogram of UCH-L1, synaptopodin, CD2AP, nephrin, and Snail protein expression. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 both versus control group and control infection group.
Mentions: To further investigate the effect of increased UCH-L1 in podocytes, several important molecules of podocytes were assessed after differentiated podocytes were infected with UCH-L1 overexpression adenoviral vector for 48 h. As shown in Figure 7, expression of UCH-L1 in UCH-L1 infected-group was significantly high than that of control group and control infection group, which demonstrated the successful infection. The successful infection decreased the expression of synaptopodin, CD2AP and nephrin and increased that of Snail. These results demonstrated that UCH-L1 could change the expression of important proteins in podocytes, which may correspondingly affect its structure and function.

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