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Effect of berberine on the renal tubular epithelial-to-mesenchymal transition by inhibition of the Notch/snail pathway in diabetic nephropathy model KKAy mice

View Article: PubMed Central - PubMed

ABSTRACT

Renal tubular epithelial-to-mesenchymal transition (EMT) and renal tubular interstitial fibrosis are the main pathological changes of diabetic nephropathy (DN), which is a common cause of end-stage renal disease. Previous studies have suggested that berberine (BBR) has antifibrotic effects in the kidney and can reduce apoptosis and inhibit the EMT of podocytes in DN. However, the effect of BBR on the renal tubular EMT in DN and its mechanisms of action are unknown. This study was performed to explore the effects of BBR on the renal tubular EMT and the molecular mechanisms of BBR in DN model KKAy mice and on the high glucose (HG)-induced EMT in mouse renal tubular epithelial cells. Our results showed that, relative to the model mice, the mice in the treatment group had an improved general state and reduced blood glucose and 24-h urinary protein levels. Degradation of renal function was ameliorated by BBR. We also observed the protective effects of BBR on renal structural changes, including normalization of an index of renal interstitial fibrosis and kidney weight/body weight. Moreover, BBR suppressed the activation of the Notch/snail pathway and upregulated the α-SMA and E-cadherin levels in DN model KKAy mice. BBR was further found to prevent HG-induced EMT events and to inhibit the HG-induced expression of Notch pathway members and snail1 in mouse renal tubular epithelial cells. Our findings indicate that BBR has a therapeutic effect on DN, including its inhibition of the renal tubular EMT and renal interstitial fibrosis. Furthermore, the BBR-mediated EMT inhibition occurs through Notch/snail pathway regulation.

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Berberine suppresses renal tubular epithelial-to-mesenchymal transition in DN model KKAy mice by upregulating the expression of E-Cad and downregulating the expression of α-SMA.Notes: (A) Representative photographs of immunohistochemistry for E-Cad and α-SMA in DN model KKAy mice, 400×. (B) Representative band of E-Cad and α-SMA proteins by Western blot in DN model KKAy mice. (C) Comparison of MOD of E-Cad and α-SMA proteins in DN model KKAy mice. (D) Comparison of the gray value of E-Cad and α-SMA proteins in DN model KKAy mice (n=3). (E) Representative photographs of in situ hybridization for E-Cad and α-SMA in DN model KKAy mice, 400×. (F) Comparison of MOD of E-Cad and α-SMA mRNA in DN model KKAy mice. **P<0.01, NG and ##P<0.01.Abbreviations: DN, diabetic nephropathy; E-Cad, E-cadherin; MG, model group; MOD, mean optical density; NG, normal control group; TG, treatment group.
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f5-dddt-11-1065: Berberine suppresses renal tubular epithelial-to-mesenchymal transition in DN model KKAy mice by upregulating the expression of E-Cad and downregulating the expression of α-SMA.Notes: (A) Representative photographs of immunohistochemistry for E-Cad and α-SMA in DN model KKAy mice, 400×. (B) Representative band of E-Cad and α-SMA proteins by Western blot in DN model KKAy mice. (C) Comparison of MOD of E-Cad and α-SMA proteins in DN model KKAy mice. (D) Comparison of the gray value of E-Cad and α-SMA proteins in DN model KKAy mice (n=3). (E) Representative photographs of in situ hybridization for E-Cad and α-SMA in DN model KKAy mice, 400×. (F) Comparison of MOD of E-Cad and α-SMA mRNA in DN model KKAy mice. **P<0.01, NG and ##P<0.01.Abbreviations: DN, diabetic nephropathy; E-Cad, E-cadherin; MG, model group; MOD, mean optical density; NG, normal control group; TG, treatment group.

Mentions: It is established that E-Cad is a marker of epithelial cells, while α-SMA is a mesenchymal cell marker. The EMT is a process in which renal tubular epithelial cells lose their epithelial markers, change their morphology, become muscle fibroblasts, and obtain the mesenchymal cell marker α-SMA, to combat injury and avoid potential apoptosis. To investigate the effects of BBR on inhibition of the EMT in DN model KKAy mice, E-Cad and α-SMA protein expression was detected by Western blot and immunohistochemistry (Figure 5A and B). Relative to the normal group, E-Cad expression was markedly decreased in the DN model KKAy mice (P<0.01; Figure 5C and D), while α-SMA expression was increased (P<0.01; Figure 5B and D). BBR attenuated the decreased α-SMA expression in the DN model KKAy mice (P<0.01; Figure 5B and D). Moreover, E-Cad and α-SMA mRNA expression levels were detected by ISH (Figure 5E), consistent with the results from Western blot and immunohistochemistry. The E-Cad mRNA expression level was significantly decreased in the MG relative to the NG (P<0.01; Figure 5F). Moreover, the α-SMA mRNA expression was also significantly increased in the DN model KKAy mice (P<0.01; Figure 5F). BBR reversed the decreased α-SMA protein expression and mRNA transcription level and increased the E-Cad protein expression and mRNA transcription level in the DN model KKAy mice (P<0.01; Figure 5F), which suggests that BBR can suppress the renal tubular EMT in DN model KKAy mice.


Effect of berberine on the renal tubular epithelial-to-mesenchymal transition by inhibition of the Notch/snail pathway in diabetic nephropathy model KKAy mice
Berberine suppresses renal tubular epithelial-to-mesenchymal transition in DN model KKAy mice by upregulating the expression of E-Cad and downregulating the expression of α-SMA.Notes: (A) Representative photographs of immunohistochemistry for E-Cad and α-SMA in DN model KKAy mice, 400×. (B) Representative band of E-Cad and α-SMA proteins by Western blot in DN model KKAy mice. (C) Comparison of MOD of E-Cad and α-SMA proteins in DN model KKAy mice. (D) Comparison of the gray value of E-Cad and α-SMA proteins in DN model KKAy mice (n=3). (E) Representative photographs of in situ hybridization for E-Cad and α-SMA in DN model KKAy mice, 400×. (F) Comparison of MOD of E-Cad and α-SMA mRNA in DN model KKAy mice. **P<0.01, NG and ##P<0.01.Abbreviations: DN, diabetic nephropathy; E-Cad, E-cadherin; MG, model group; MOD, mean optical density; NG, normal control group; TG, treatment group.
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f5-dddt-11-1065: Berberine suppresses renal tubular epithelial-to-mesenchymal transition in DN model KKAy mice by upregulating the expression of E-Cad and downregulating the expression of α-SMA.Notes: (A) Representative photographs of immunohistochemistry for E-Cad and α-SMA in DN model KKAy mice, 400×. (B) Representative band of E-Cad and α-SMA proteins by Western blot in DN model KKAy mice. (C) Comparison of MOD of E-Cad and α-SMA proteins in DN model KKAy mice. (D) Comparison of the gray value of E-Cad and α-SMA proteins in DN model KKAy mice (n=3). (E) Representative photographs of in situ hybridization for E-Cad and α-SMA in DN model KKAy mice, 400×. (F) Comparison of MOD of E-Cad and α-SMA mRNA in DN model KKAy mice. **P<0.01, NG and ##P<0.01.Abbreviations: DN, diabetic nephropathy; E-Cad, E-cadherin; MG, model group; MOD, mean optical density; NG, normal control group; TG, treatment group.
Mentions: It is established that E-Cad is a marker of epithelial cells, while α-SMA is a mesenchymal cell marker. The EMT is a process in which renal tubular epithelial cells lose their epithelial markers, change their morphology, become muscle fibroblasts, and obtain the mesenchymal cell marker α-SMA, to combat injury and avoid potential apoptosis. To investigate the effects of BBR on inhibition of the EMT in DN model KKAy mice, E-Cad and α-SMA protein expression was detected by Western blot and immunohistochemistry (Figure 5A and B). Relative to the normal group, E-Cad expression was markedly decreased in the DN model KKAy mice (P<0.01; Figure 5C and D), while α-SMA expression was increased (P<0.01; Figure 5B and D). BBR attenuated the decreased α-SMA expression in the DN model KKAy mice (P<0.01; Figure 5B and D). Moreover, E-Cad and α-SMA mRNA expression levels were detected by ISH (Figure 5E), consistent with the results from Western blot and immunohistochemistry. The E-Cad mRNA expression level was significantly decreased in the MG relative to the NG (P<0.01; Figure 5F). Moreover, the α-SMA mRNA expression was also significantly increased in the DN model KKAy mice (P<0.01; Figure 5F). BBR reversed the decreased α-SMA protein expression and mRNA transcription level and increased the E-Cad protein expression and mRNA transcription level in the DN model KKAy mice (P<0.01; Figure 5F), which suggests that BBR can suppress the renal tubular EMT in DN model KKAy mice.

View Article: PubMed Central - PubMed

ABSTRACT

Renal tubular epithelial-to-mesenchymal transition (EMT) and renal tubular interstitial fibrosis are the main pathological changes of diabetic nephropathy (DN), which is a common cause of end-stage renal disease. Previous studies have suggested that berberine (BBR) has antifibrotic effects in the kidney and can reduce apoptosis and inhibit the EMT of podocytes in DN. However, the effect of BBR on the renal tubular EMT in DN and its mechanisms of action are unknown. This study was performed to explore the effects of BBR on the renal tubular EMT and the molecular mechanisms of BBR in DN model KKAy mice and on the high glucose (HG)-induced EMT in mouse renal tubular epithelial cells. Our results showed that, relative to the model mice, the mice in the treatment group had an improved general state and reduced blood glucose and 24-h urinary protein levels. Degradation of renal function was ameliorated by BBR. We also observed the protective effects of BBR on renal structural changes, including normalization of an index of renal interstitial fibrosis and kidney weight/body weight. Moreover, BBR suppressed the activation of the Notch/snail pathway and upregulated the &alpha;-SMA and E-cadherin levels in DN model KKAy mice. BBR was further found to prevent HG-induced EMT events and to inhibit the HG-induced expression of Notch pathway members and snail1 in mouse renal tubular epithelial cells. Our findings indicate that BBR has a therapeutic effect on DN, including its inhibition of the renal tubular EMT and renal interstitial fibrosis. Furthermore, the BBR-mediated EMT inhibition occurs through Notch/snail pathway regulation.

No MeSH data available.


Related in: MedlinePlus