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Hydroxysafflor Yellow A Ameliorates Renal Fibrosis by Suppressing TGF-β1-Induced Epithelial-to-Mesenchymal Transition.

Hu N, Duan J, Li H, Wang Y, Wang F, Chu J, Sun J, Liu M, Wang C, Lu C, Wen A - PLoS ONE (2016)

Bottom Line: HE staining and Masson staining demonstrated that kidney interstitial fibrosis, tubular atrophy, and inflammatory cell infiltration were notably attenuated in the high-dose HSYA group compared with the model group.The expressions of α-SMA, collagen-I and fibronectin were decreased in the UUO kidney and HK-2 cells of the HSYA-treatment group.Moreover, HSYA reduced the apoptotic rate of HK-2 cells stimulated by TGF-β1.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.

ABSTRACT

Objective: Renal fibrosis is the common pathological foundation of many chronic kidney diseases (CKDs). The aim of this study was to investigate whether Hydroxysafflor yellow A (HSYA) can preserve renal function by inhibiting the progression of renal fibrosis and the potential mechanisms.

Methods: Renal fibrosis was induced by unilateral ureteral obstruction (UUO) performed on 7-week-old C57BL/6 mice. HSYA (10, 50 and 100 mg/kg) were intragastrically administered. Sham group and model group were administered with the same volume of vehicle. Serum and kidney samples were collected 14 days after the UUO surgery. Serum biochemical indicators were measured by automatic biochemical analyzer. Histological changes were evaluated by HE and Masson staining. In vitro, the anti-fibrotic effect of HSYA was tested on human recombinant transforming growth factor-β1 (TGF-β1) stimulated HK-2 cells. The protein levels of α-SMA, collagen-I and fibronectin in kidney tissue and HK-2 cells were measured by immunohistochemistry and immunofluorescence. The protein levels of apoptosis-relative and TGF-β1/Smad3 signaling were detected by western blot.

Results: HSYA slowed the development of renal fibrosis both in vivo and in vitro. In UUO rats, renal function index suggested that HSYA treatment decreased the level of serum creatinine (Scr) and blood urea nitrogen (BUN) rose by UUO (P<0.05). HE staining and Masson staining demonstrated that kidney interstitial fibrosis, tubular atrophy, and inflammatory cell infiltration were notably attenuated in the high-dose HSYA group compared with the model group. The expressions of α-SMA, collagen-I and fibronectin were decreased in the UUO kidney and HK-2 cells of the HSYA-treatment group. Moreover, HSYA reduced the apoptotic rate of HK-2 cells stimulated by TGF-β1. Further study revealed that HSYA regulated the TGF-β1/Smads signaling pathway both in kidney tissue and HK-2 cells.

Conclusions: These results suggested that HSYA had a protective effect against fibrosis in renal cells, at least partly, through inhibiting TGF-β1/smad3-mediated Epithelial-mesenchymal transition signaling pathway.

No MeSH data available.


Related in: MedlinePlus

Effect of HSYA on TGF-β1/Smads signaling pathway in HK-2 cells.Western blotting was performed to detect TGF-β1, Smad3, p-Smad3, Smad7 protein levels, and β-actin was used as a loading control (A). Semi-quantitative data from densitometric analysis of TGF-β1 and Smad7 are presented as relative ratio of each protein to β-actin, p-Smad3 are presented as the ratio to Smad3 (B-D). Data are expressed as mean±SD, n = 3, #P<0.05,##P<0.01compared with control group; *P<0.05 compared with TGF-β1-stimulated group.
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pone.0153409.g008: Effect of HSYA on TGF-β1/Smads signaling pathway in HK-2 cells.Western blotting was performed to detect TGF-β1, Smad3, p-Smad3, Smad7 protein levels, and β-actin was used as a loading control (A). Semi-quantitative data from densitometric analysis of TGF-β1 and Smad7 are presented as relative ratio of each protein to β-actin, p-Smad3 are presented as the ratio to Smad3 (B-D). Data are expressed as mean±SD, n = 3, #P<0.05,##P<0.01compared with control group; *P<0.05 compared with TGF-β1-stimulated group.

Mentions: We then test the effect of HSYA on TGF-β1/Smad3 signaling, which have been confirmed a central role to regulate interstitial fibrosis [22, 23].The level of TGF-β1 was down-regulated compare with the model group, and the ratio of p-Smad3 to Smad3 was decreased in the HSYA-treated groups relative to the model group in a dose-dependent manner (Fig 7 and S3 Table). The semblable effect was identified in HK-2cells. Administration with HSYA notably reduced the high expression of TGF-β1 and p-Smad3 stimulated by TGF-β1. In addition, the negative regulator, Smad7 was up-regulated by HSYA (Fig 8). These results suggested that activation of the TGF-β1/Smads signaling pathway plays a prominent role in the progress of renal fibrosis and it was meaningful that HSYA could rebalance the TGF-β1/Smad ssignaling to exhibiting antifibrotic effect.


Hydroxysafflor Yellow A Ameliorates Renal Fibrosis by Suppressing TGF-β1-Induced Epithelial-to-Mesenchymal Transition.

Hu N, Duan J, Li H, Wang Y, Wang F, Chu J, Sun J, Liu M, Wang C, Lu C, Wen A - PLoS ONE (2016)

Effect of HSYA on TGF-β1/Smads signaling pathway in HK-2 cells.Western blotting was performed to detect TGF-β1, Smad3, p-Smad3, Smad7 protein levels, and β-actin was used as a loading control (A). Semi-quantitative data from densitometric analysis of TGF-β1 and Smad7 are presented as relative ratio of each protein to β-actin, p-Smad3 are presented as the ratio to Smad3 (B-D). Data are expressed as mean±SD, n = 3, #P<0.05,##P<0.01compared with control group; *P<0.05 compared with TGF-β1-stimulated group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835075&req=5

pone.0153409.g008: Effect of HSYA on TGF-β1/Smads signaling pathway in HK-2 cells.Western blotting was performed to detect TGF-β1, Smad3, p-Smad3, Smad7 protein levels, and β-actin was used as a loading control (A). Semi-quantitative data from densitometric analysis of TGF-β1 and Smad7 are presented as relative ratio of each protein to β-actin, p-Smad3 are presented as the ratio to Smad3 (B-D). Data are expressed as mean±SD, n = 3, #P<0.05,##P<0.01compared with control group; *P<0.05 compared with TGF-β1-stimulated group.
Mentions: We then test the effect of HSYA on TGF-β1/Smad3 signaling, which have been confirmed a central role to regulate interstitial fibrosis [22, 23].The level of TGF-β1 was down-regulated compare with the model group, and the ratio of p-Smad3 to Smad3 was decreased in the HSYA-treated groups relative to the model group in a dose-dependent manner (Fig 7 and S3 Table). The semblable effect was identified in HK-2cells. Administration with HSYA notably reduced the high expression of TGF-β1 and p-Smad3 stimulated by TGF-β1. In addition, the negative regulator, Smad7 was up-regulated by HSYA (Fig 8). These results suggested that activation of the TGF-β1/Smads signaling pathway plays a prominent role in the progress of renal fibrosis and it was meaningful that HSYA could rebalance the TGF-β1/Smad ssignaling to exhibiting antifibrotic effect.

Bottom Line: HE staining and Masson staining demonstrated that kidney interstitial fibrosis, tubular atrophy, and inflammatory cell infiltration were notably attenuated in the high-dose HSYA group compared with the model group.The expressions of α-SMA, collagen-I and fibronectin were decreased in the UUO kidney and HK-2 cells of the HSYA-treatment group.Moreover, HSYA reduced the apoptotic rate of HK-2 cells stimulated by TGF-β1.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.

ABSTRACT

Objective: Renal fibrosis is the common pathological foundation of many chronic kidney diseases (CKDs). The aim of this study was to investigate whether Hydroxysafflor yellow A (HSYA) can preserve renal function by inhibiting the progression of renal fibrosis and the potential mechanisms.

Methods: Renal fibrosis was induced by unilateral ureteral obstruction (UUO) performed on 7-week-old C57BL/6 mice. HSYA (10, 50 and 100 mg/kg) were intragastrically administered. Sham group and model group were administered with the same volume of vehicle. Serum and kidney samples were collected 14 days after the UUO surgery. Serum biochemical indicators were measured by automatic biochemical analyzer. Histological changes were evaluated by HE and Masson staining. In vitro, the anti-fibrotic effect of HSYA was tested on human recombinant transforming growth factor-β1 (TGF-β1) stimulated HK-2 cells. The protein levels of α-SMA, collagen-I and fibronectin in kidney tissue and HK-2 cells were measured by immunohistochemistry and immunofluorescence. The protein levels of apoptosis-relative and TGF-β1/Smad3 signaling were detected by western blot.

Results: HSYA slowed the development of renal fibrosis both in vivo and in vitro. In UUO rats, renal function index suggested that HSYA treatment decreased the level of serum creatinine (Scr) and blood urea nitrogen (BUN) rose by UUO (P<0.05). HE staining and Masson staining demonstrated that kidney interstitial fibrosis, tubular atrophy, and inflammatory cell infiltration were notably attenuated in the high-dose HSYA group compared with the model group. The expressions of α-SMA, collagen-I and fibronectin were decreased in the UUO kidney and HK-2 cells of the HSYA-treatment group. Moreover, HSYA reduced the apoptotic rate of HK-2 cells stimulated by TGF-β1. Further study revealed that HSYA regulated the TGF-β1/Smads signaling pathway both in kidney tissue and HK-2 cells.

Conclusions: These results suggested that HSYA had a protective effect against fibrosis in renal cells, at least partly, through inhibiting TGF-β1/smad3-mediated Epithelial-mesenchymal transition signaling pathway.

No MeSH data available.


Related in: MedlinePlus