Limits...
Effects of Chinese Medicine Tong xinluo on Diabetic Nephropathy via Inhibiting TGF- β 1-Induced Epithelial-to-Mesenchymal Transition.

Zhang N, Gao Y, Zou D, Wang J, Li J, Zhou S, Zhu Z, Zhao X, Xu L, Zhang H - Evid Based Complement Alternat Med (2014)

Bottom Line: As a result, the expressions of collagen IV (Col IV) and fibronectin (FN) were significantly decreased in TXL group.In vivo, 24 h-UAER (24-hour urine albumin excretion ratio) and BUN (blood urea nitrogen) were decreased and Ccr (creatinine clearance ratio) was increased in TXL group compared with DN group.In summary, the present study demonstrates that TXL successfully inhibits TGF- β 1-induced epithelial-to-mesenchymal transition in DN, which may account for the therapeutic efficacy in TXL-mediated renoprotection.

View Article: PubMed Central - PubMed

Affiliation: School of Traditional Chinese Medicine, Capital Medical University, No. 10, Youanmenwai, Xitoutiao, Fengtai District, Beijing 100069, China.

ABSTRACT
Diabetic nephropathy (DN) is a major cause of chronic kidney failure and characterized by interstitial and glomeruli fibrosis. Epithelial-to-mesenchymal transition (EMT) plays an important role in the pathogenesis of DN. Tong xinluo (TXL), a Chinese herbal compound, has been used in China with established therapeutic efficacy in patients with DN. To investigate the molecular mechanism of TXL improving DN, KK-Ay mice were selected as models for the evaluation of pathogenesis and treatment in DN. In vitro, TGF- β 1 was used to induce EMT. Western blot (WB), immunofluorescence staining, and real-time polymerase chain reaction (RT-PCR) were applied to detect the changes of EMT markers in vivo and in vitro, respectively. Results showed the expressions of TGF- β 1 and its downstream proteins smad3/p-smad3 were greatly reduced in TXL group; meantime, TXL restored the expression of smad7. As a result, the expressions of collagen IV (Col IV) and fibronectin (FN) were significantly decreased in TXL group. In vivo, 24 h-UAER (24-hour urine albumin excretion ratio) and BUN (blood urea nitrogen) were decreased and Ccr (creatinine clearance ratio) was increased in TXL group compared with DN group. In summary, the present study demonstrates that TXL successfully inhibits TGF- β 1-induced epithelial-to-mesenchymal transition in DN, which may account for the therapeutic efficacy in TXL-mediated renoprotection.

No MeSH data available.


Related in: MedlinePlus

Effects of TXL on the expressions of p-smad3/smad3 and smad7 in both renal tissues and HKCs. (a) Representative bands of p-smad3/smad3 and smad7 detected by western blot of renal tissues. (b) Densitometry analysis of p-smad3/smad3 and smad7 bands from (a), normalized to GAPDH. (c) Representative bands of p-smad3/smad3 and smad7 detected by western blot of HKCs. (d) Densitometry analysis of p-smad3/smad3 and smad7 bands from (b), normalized to GAPDH. (e) Effects of TXL on smad3 and smad7 mRNA expressions in renal tissues, normalized to β-actin. (f) Effects of TXL on smad3 and smad7 mRNA expressions in HKCs, normalized to β-actin.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4016864&req=5

fig4: Effects of TXL on the expressions of p-smad3/smad3 and smad7 in both renal tissues and HKCs. (a) Representative bands of p-smad3/smad3 and smad7 detected by western blot of renal tissues. (b) Densitometry analysis of p-smad3/smad3 and smad7 bands from (a), normalized to GAPDH. (c) Representative bands of p-smad3/smad3 and smad7 detected by western blot of HKCs. (d) Densitometry analysis of p-smad3/smad3 and smad7 bands from (b), normalized to GAPDH. (e) Effects of TXL on smad3 and smad7 mRNA expressions in renal tissues, normalized to β-actin. (f) Effects of TXL on smad3 and smad7 mRNA expressions in HKCs, normalized to β-actin.

Mentions: In EMT, various studies have explored the roles ofTGF-β1 in activating smads [31]. To explore whether TXL affects TGF-β1/smads signal pathway, renal tissue and HKCs were detected by RT-PCR and western blot, respectively. 10 ng/mL TGF-β1 was applied to induce EMT in HKCs. Cells exposed to 10 ng/mL TGF-β1 were set as TGF-β1 group, and cells exposed to 10 ng/mL TGF-β1 and 250 μg/mL TXL were set as TGF-β1 + TXL group. The normal cells cultured only with DMEM/F12 were set as normal group. In vivo, western blot showed that p-smad3/smad3 were remarkably increased and smad7 was decreased in DN group compared with normal group (Figures 4(a) and 4(b), *P < 0.05), while p-smad3 and smad3 were significantly decreased and smad7 was increased in TXL group compared with DN group (Figures 4(a) and 4(b), ▲P < 0.05). RT-PCR results were consistent with western blot results (Figure 4(e)). Additionally, in vitro, western blot and RT-PCR also showed the significant increase of p-smad3 and smad3 in TGF-β1 group compared with normal group. In contrast, smad7 was significantly decreased (Figures 4(c), 4(d), and 4(f), *P < 0.05). Importantly, p-smad3/smad3 were remarkably decreased and smad7 was increased in the TGF-β1 + TXL group compared with TGF-β1 group (Figures 4(c), 4(d), and 4(f), ▲P < 0.05). Thus, the present results showed that TXL positively deceased p-smad3/smad3 expressions and increased the expression of smad7.


Effects of Chinese Medicine Tong xinluo on Diabetic Nephropathy via Inhibiting TGF- β 1-Induced Epithelial-to-Mesenchymal Transition.

Zhang N, Gao Y, Zou D, Wang J, Li J, Zhou S, Zhu Z, Zhao X, Xu L, Zhang H - Evid Based Complement Alternat Med (2014)

Effects of TXL on the expressions of p-smad3/smad3 and smad7 in both renal tissues and HKCs. (a) Representative bands of p-smad3/smad3 and smad7 detected by western blot of renal tissues. (b) Densitometry analysis of p-smad3/smad3 and smad7 bands from (a), normalized to GAPDH. (c) Representative bands of p-smad3/smad3 and smad7 detected by western blot of HKCs. (d) Densitometry analysis of p-smad3/smad3 and smad7 bands from (b), normalized to GAPDH. (e) Effects of TXL on smad3 and smad7 mRNA expressions in renal tissues, normalized to β-actin. (f) Effects of TXL on smad3 and smad7 mRNA expressions in HKCs, normalized to β-actin.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4016864&req=5

fig4: Effects of TXL on the expressions of p-smad3/smad3 and smad7 in both renal tissues and HKCs. (a) Representative bands of p-smad3/smad3 and smad7 detected by western blot of renal tissues. (b) Densitometry analysis of p-smad3/smad3 and smad7 bands from (a), normalized to GAPDH. (c) Representative bands of p-smad3/smad3 and smad7 detected by western blot of HKCs. (d) Densitometry analysis of p-smad3/smad3 and smad7 bands from (b), normalized to GAPDH. (e) Effects of TXL on smad3 and smad7 mRNA expressions in renal tissues, normalized to β-actin. (f) Effects of TXL on smad3 and smad7 mRNA expressions in HKCs, normalized to β-actin.
Mentions: In EMT, various studies have explored the roles ofTGF-β1 in activating smads [31]. To explore whether TXL affects TGF-β1/smads signal pathway, renal tissue and HKCs were detected by RT-PCR and western blot, respectively. 10 ng/mL TGF-β1 was applied to induce EMT in HKCs. Cells exposed to 10 ng/mL TGF-β1 were set as TGF-β1 group, and cells exposed to 10 ng/mL TGF-β1 and 250 μg/mL TXL were set as TGF-β1 + TXL group. The normal cells cultured only with DMEM/F12 were set as normal group. In vivo, western blot showed that p-smad3/smad3 were remarkably increased and smad7 was decreased in DN group compared with normal group (Figures 4(a) and 4(b), *P < 0.05), while p-smad3 and smad3 were significantly decreased and smad7 was increased in TXL group compared with DN group (Figures 4(a) and 4(b), ▲P < 0.05). RT-PCR results were consistent with western blot results (Figure 4(e)). Additionally, in vitro, western blot and RT-PCR also showed the significant increase of p-smad3 and smad3 in TGF-β1 group compared with normal group. In contrast, smad7 was significantly decreased (Figures 4(c), 4(d), and 4(f), *P < 0.05). Importantly, p-smad3/smad3 were remarkably decreased and smad7 was increased in the TGF-β1 + TXL group compared with TGF-β1 group (Figures 4(c), 4(d), and 4(f), ▲P < 0.05). Thus, the present results showed that TXL positively deceased p-smad3/smad3 expressions and increased the expression of smad7.

Bottom Line: As a result, the expressions of collagen IV (Col IV) and fibronectin (FN) were significantly decreased in TXL group.In vivo, 24 h-UAER (24-hour urine albumin excretion ratio) and BUN (blood urea nitrogen) were decreased and Ccr (creatinine clearance ratio) was increased in TXL group compared with DN group.In summary, the present study demonstrates that TXL successfully inhibits TGF- β 1-induced epithelial-to-mesenchymal transition in DN, which may account for the therapeutic efficacy in TXL-mediated renoprotection.

View Article: PubMed Central - PubMed

Affiliation: School of Traditional Chinese Medicine, Capital Medical University, No. 10, Youanmenwai, Xitoutiao, Fengtai District, Beijing 100069, China.

ABSTRACT
Diabetic nephropathy (DN) is a major cause of chronic kidney failure and characterized by interstitial and glomeruli fibrosis. Epithelial-to-mesenchymal transition (EMT) plays an important role in the pathogenesis of DN. Tong xinluo (TXL), a Chinese herbal compound, has been used in China with established therapeutic efficacy in patients with DN. To investigate the molecular mechanism of TXL improving DN, KK-Ay mice were selected as models for the evaluation of pathogenesis and treatment in DN. In vitro, TGF- β 1 was used to induce EMT. Western blot (WB), immunofluorescence staining, and real-time polymerase chain reaction (RT-PCR) were applied to detect the changes of EMT markers in vivo and in vitro, respectively. Results showed the expressions of TGF- β 1 and its downstream proteins smad3/p-smad3 were greatly reduced in TXL group; meantime, TXL restored the expression of smad7. As a result, the expressions of collagen IV (Col IV) and fibronectin (FN) were significantly decreased in TXL group. In vivo, 24 h-UAER (24-hour urine albumin excretion ratio) and BUN (blood urea nitrogen) were decreased and Ccr (creatinine clearance ratio) was increased in TXL group compared with DN group. In summary, the present study demonstrates that TXL successfully inhibits TGF- β 1-induced epithelial-to-mesenchymal transition in DN, which may account for the therapeutic efficacy in TXL-mediated renoprotection.

No MeSH data available.


Related in: MedlinePlus