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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-induced Apoptosis in HK-2 Cells.The apoptosis-relative protein level of Bax, Bcl-2 and cleaved caspase3 were measured. Semi-quantitative data from densitometric analysis of Bax, Bcl-2 and caspase3 are presented as relative ratio of each protein to β-actin. Data are expressed as mean±SD, n = 3, #P<0.05 compared with control group; *P<0.05 compared with TGF-β1-stimulated group.
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pone.0153409.g006: Effect of HSYA on TGF-β1-induced Apoptosis in HK-2 Cells.The apoptosis-relative protein level of Bax, Bcl-2 and cleaved caspase3 were measured. Semi-quantitative data from densitometric analysis of Bax, Bcl-2 and caspase3 are presented as relative ratio of each protein to β-actin. Data are expressed as mean±SD, n = 3, #P<0.05 compared with control group; *P<0.05 compared with TGF-β1-stimulated group.

Mentions: It is well-known that cell apoptosis involved in the injury and repair progress of multiple kidney disease. Growing evidence showed that TGF-β1/Smads signaling could mediate cell apoptosis by stimulating expression of pro-apoptotic members and by suppressing expression of anti-apoptotic members, respectively [21]. Here, we test the expression of pro-apoptotic Bax, anti-apoptotic Bcl-2, and cleaved caspase3 by Western blotting in HK-2 cells. Data (S5 Table) indicated that Bax and caspase3 protein exhibited higher expression in TGF-β1-stimulated group. While, Bcl-2 expression was lower in TGF-β1-stimulated group than in control group. The expression of Bax, Bcl-2 and caspase3 were all reversed in HSYA group. These phenomena suggested that HSYA could reduce cell apoptosis via regulating the ratio of Bax and Bcl-2, down-regulating the expression of cleaved caspase3. (Fig 6)


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-induced Apoptosis in HK-2 Cells.The apoptosis-relative protein level of Bax, Bcl-2 and cleaved caspase3 were measured. Semi-quantitative data from densitometric analysis of Bax, Bcl-2 and caspase3 are presented as relative ratio of each protein to β-actin. Data are expressed as mean±SD, n = 3, #P<0.05 compared with control group; *P<0.05 compared with TGF-β1-stimulated group.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835075&req=5

pone.0153409.g006: Effect of HSYA on TGF-β1-induced Apoptosis in HK-2 Cells.The apoptosis-relative protein level of Bax, Bcl-2 and cleaved caspase3 were measured. Semi-quantitative data from densitometric analysis of Bax, Bcl-2 and caspase3 are presented as relative ratio of each protein to β-actin. Data are expressed as mean±SD, n = 3, #P<0.05 compared with control group; *P<0.05 compared with TGF-β1-stimulated group.
Mentions: It is well-known that cell apoptosis involved in the injury and repair progress of multiple kidney disease. Growing evidence showed that TGF-β1/Smads signaling could mediate cell apoptosis by stimulating expression of pro-apoptotic members and by suppressing expression of anti-apoptotic members, respectively [21]. Here, we test the expression of pro-apoptotic Bax, anti-apoptotic Bcl-2, and cleaved caspase3 by Western blotting in HK-2 cells. Data (S5 Table) indicated that Bax and caspase3 protein exhibited higher expression in TGF-β1-stimulated group. While, Bcl-2 expression was lower in TGF-β1-stimulated group than in control group. The expression of Bax, Bcl-2 and caspase3 were all reversed in HSYA group. These phenomena suggested that HSYA could reduce cell apoptosis via regulating the ratio of Bax and Bcl-2, down-regulating the expression of cleaved caspase3. (Fig 6)

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