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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.


Activation of the Wnt/β-catenin pathway induced by high glucose in podocytes Podocytes incubated with HG for 24 and 48 h, respectively. (A) Reverse transcription-PCR (RT-PCR) demonstrating an non-altered expression of Wnt1 and Wnt2 and an increased expression of Wnt5a and β-catenin compared to control; (B) corresponding statistic histogram of Wnt1, Wnt2, Wnt5a and β-catenin. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 compared to control group and HG 24 h group respectively.
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ijms-17-01404-f002: Activation of the Wnt/β-catenin pathway induced by high glucose in podocytes Podocytes incubated with HG for 24 and 48 h, respectively. (A) Reverse transcription-PCR (RT-PCR) demonstrating an non-altered expression of Wnt1 and Wnt2 and an increased expression of Wnt5a and β-catenin compared to control; (B) corresponding statistic histogram of Wnt1, Wnt2, Wnt5a and β-catenin. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 compared to control group and HG 24 h group respectively.

Mentions: An analysis of the mRNA expression of common measured Wnt genes was detected by reverse transcription-PCR (RT-PCR) approach to explore the activation status of Wnt/β-catenin signaling pathway in the control group and the HG group. As shown in Figure 2, compared to control, mRNA expression of Wnt1 and Wnt2 were not markedly changed in the HG group, neither by 24 h nor by 48 h. However, after HG treatment, mRNA expression of Wnt5a was evidently increased by 48 h, ~3.1 folds.


Wnt/ β -Catenin Signaling Mediated-UCH-L1 Expression in Podocytes of Diabetic Nephropathy
Activation of the Wnt/β-catenin pathway induced by high glucose in podocytes Podocytes incubated with HG for 24 and 48 h, respectively. (A) Reverse transcription-PCR (RT-PCR) demonstrating an non-altered expression of Wnt1 and Wnt2 and an increased expression of Wnt5a and β-catenin compared to control; (B) corresponding statistic histogram of Wnt1, Wnt2, Wnt5a and β-catenin. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 compared to control group and HG 24 h group respectively.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-17-01404-f002: Activation of the Wnt/β-catenin pathway induced by high glucose in podocytes Podocytes incubated with HG for 24 and 48 h, respectively. (A) Reverse transcription-PCR (RT-PCR) demonstrating an non-altered expression of Wnt1 and Wnt2 and an increased expression of Wnt5a and β-catenin compared to control; (B) corresponding statistic histogram of Wnt1, Wnt2, Wnt5a and β-catenin. Data representative of three independent experiments. * p < 0.05, ** p < 0.01 compared to control group and HG 24 h group respectively.
Mentions: An analysis of the mRNA expression of common measured Wnt genes was detected by reverse transcription-PCR (RT-PCR) approach to explore the activation status of Wnt/β-catenin signaling pathway in the control group and the HG group. As shown in Figure 2, compared to control, mRNA expression of Wnt1 and Wnt2 were not markedly changed in the HG group, neither by 24 h nor by 48 h. However, after HG treatment, mRNA expression of Wnt5a was evidently increased by 48 h, ~3.1 folds.

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.