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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.

No MeSH data available.


BBR inhibits the HG-induced expression of snail1.Notes: (A) Representative band of snail1 protein by Western blot in mRTECs. (B) Comparison of the gray value of snail1 protein in mRTECs (n=3). (C) Comparison of the mRNA level of snail1 protein by reverse transcription polymerase chain reaction in mRTECs (n=3). **P<0.01 NG and ##P<0.01, #P<0.05 compared with the model group.Abbreviations: BBR, berberine; DAPT, N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester; HG, high glucose; mRTECs, mouse renal tubular epithelial cells; NG, normal control group.
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f10-dddt-11-1065: BBR inhibits the HG-induced expression of snail1.Notes: (A) Representative band of snail1 protein by Western blot in mRTECs. (B) Comparison of the gray value of snail1 protein in mRTECs (n=3). (C) Comparison of the mRNA level of snail1 protein by reverse transcription polymerase chain reaction in mRTECs (n=3). **P<0.01 NG and ##P<0.01, #P<0.05 compared with the model group.Abbreviations: BBR, berberine; DAPT, N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester; HG, high glucose; mRTECs, mouse renal tubular epithelial cells; NG, normal control group.

Mentions: Similarly, snail1 protein and mRNA expression of mRTECs were detected by Western blot and RT-PCR, respectively, in our in vitro study. DAPT is a γ-secretase inhibitor and an indirect inhibitor of the Notch pathway. To further clarify the relationship between the Notch pathway and snail1, in the DAPT group, the cells were treated with 30 mM/L glucose after pretreatment with 10 μM/L DAPT for 40 min. Our results showed that snail1 protein and mRNA expression were significantly increased in the HG group relative to the NG (P<0.05; Figure 10A–C); also, BBR markedly inhibited snail1 protein and mRNA expression induced by HG (P<0.05; Figure 10A–C). Moreover, the protein and mRNA expression level of snail1 were also decreased in the DAPT group (P<0.05; Figure 10A–C), which implies that the snail1 expression is indeed regulated by the Notch pathway.


Effect of berberine on the renal tubular epithelial-to-mesenchymal transition by inhibition of the Notch/snail pathway in diabetic nephropathy model KKAy mice
BBR inhibits the HG-induced expression of snail1.Notes: (A) Representative band of snail1 protein by Western blot in mRTECs. (B) Comparison of the gray value of snail1 protein in mRTECs (n=3). (C) Comparison of the mRNA level of snail1 protein by reverse transcription polymerase chain reaction in mRTECs (n=3). **P<0.01 NG and ##P<0.01, #P<0.05 compared with the model group.Abbreviations: BBR, berberine; DAPT, N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester; HG, high glucose; mRTECs, mouse renal tubular epithelial cells; NG, normal control group.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5384688&req=5

f10-dddt-11-1065: BBR inhibits the HG-induced expression of snail1.Notes: (A) Representative band of snail1 protein by Western blot in mRTECs. (B) Comparison of the gray value of snail1 protein in mRTECs (n=3). (C) Comparison of the mRNA level of snail1 protein by reverse transcription polymerase chain reaction in mRTECs (n=3). **P<0.01 NG and ##P<0.01, #P<0.05 compared with the model group.Abbreviations: BBR, berberine; DAPT, N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester; HG, high glucose; mRTECs, mouse renal tubular epithelial cells; NG, normal control group.
Mentions: Similarly, snail1 protein and mRNA expression of mRTECs were detected by Western blot and RT-PCR, respectively, in our in vitro study. DAPT is a γ-secretase inhibitor and an indirect inhibitor of the Notch pathway. To further clarify the relationship between the Notch pathway and snail1, in the DAPT group, the cells were treated with 30 mM/L glucose after pretreatment with 10 μM/L DAPT for 40 min. Our results showed that snail1 protein and mRNA expression were significantly increased in the HG group relative to the NG (P<0.05; Figure 10A–C); also, BBR markedly inhibited snail1 protein and mRNA expression induced by HG (P<0.05; Figure 10A–C). Moreover, the protein and mRNA expression level of snail1 were also decreased in the DAPT group (P<0.05; Figure 10A–C), which implies that the snail1 expression is indeed regulated by the Notch pathway.

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.