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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.


UCH-L1 expression induced by high glucose in podocytes. Podocytes were maintained or not in high glucose (30 mM) for 48 h. Podocyte lysate proteins (60 μg) were analyzed respectively by western blot using murine anti-UCH-L1 antibody and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1; (B) corresponding statistic histogram of UCH-L1 protein expression; (C) quantitative real-time RT-PCR assay of UCH-L1 mRNA. Data representative of three independent experiments. * p < 0.05 compared to control group.
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ijms-17-01404-f001: UCH-L1 expression induced by high glucose in podocytes. Podocytes were maintained or not in high glucose (30 mM) for 48 h. Podocyte lysate proteins (60 μg) were analyzed respectively by western blot using murine anti-UCH-L1 antibody and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1; (B) corresponding statistic histogram of UCH-L1 protein expression; (C) quantitative real-time RT-PCR assay of UCH-L1 mRNA. Data representative of three independent experiments. * p < 0.05 compared to control group.

Mentions: The cultured podocytes were divided into the control group and the HG group which were treated with HG for 48 h. The expression of UCH-L1 in the HG group was significantly higher than that in the control one (Figure 1A,B). At the mRNA level, quantitative real time RT-PCR showed the increase of UCH-L1 in HG-treated cells were greater than that in the control group, ~4.4-fold for 48 h (Figure 1C). Overall, these results indicated that HG upregulated the expression of UCH-L1 in podocytes at both the mRNA and protein levels.


Wnt/ β -Catenin Signaling Mediated-UCH-L1 Expression in Podocytes of Diabetic Nephropathy
UCH-L1 expression induced by high glucose in podocytes. Podocytes were maintained or not in high glucose (30 mM) for 48 h. Podocyte lysate proteins (60 μg) were analyzed respectively by western blot using murine anti-UCH-L1 antibody and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1; (B) corresponding statistic histogram of UCH-L1 protein expression; (C) quantitative real-time RT-PCR assay of UCH-L1 mRNA. Data representative of three independent experiments. * p < 0.05 compared to control group.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-17-01404-f001: UCH-L1 expression induced by high glucose in podocytes. Podocytes were maintained or not in high glucose (30 mM) for 48 h. Podocyte lysate proteins (60 μg) were analyzed respectively by western blot using murine anti-UCH-L1 antibody and β-Actin was used as control for protein loading. (A) Western blot assay of UCH-L1; (B) corresponding statistic histogram of UCH-L1 protein expression; (C) quantitative real-time RT-PCR assay of UCH-L1 mRNA. Data representative of three independent experiments. * p < 0.05 compared to control group.
Mentions: The cultured podocytes were divided into the control group and the HG group which were treated with HG for 48 h. The expression of UCH-L1 in the HG group was significantly higher than that in the control one (Figure 1A,B). At the mRNA level, quantitative real time RT-PCR showed the increase of UCH-L1 in HG-treated cells were greater than that in the control group, ~4.4-fold for 48 h (Figure 1C). Overall, these results indicated that HG upregulated the expression of UCH-L1 in podocytes at both the mRNA and protein levels.

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.