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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 myofibroblasts and accumulation of extracellular matrix in HK-2 cells.Immunofluorescence staining measured the expression levels of a-SMA (A), collagen-I (B) and fibronectin (C). Western blot analysis of FN and collagen-I protein levels (D). Semi-quantitative data from densitometric analysis of FN and collagen-I are presented as relative ratio of each protein to β-actin (E). 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.g005: Effect of HSYA on TGF-β1-induced myofibroblasts and accumulation of extracellular matrix in HK-2 cells.Immunofluorescence staining measured the expression levels of a-SMA (A), collagen-I (B) and fibronectin (C). Western blot analysis of FN and collagen-I protein levels (D). Semi-quantitative data from densitometric analysis of FN and collagen-I are presented as relative ratio of each protein to β-actin (E). 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: Due to the fact that the progression of EMT paves the way for extracellular matrix (ECM) deposition and finally deteriorate renal fibrosis, we detected the expression of a-SMA, collagen-I and fibronectin by Immunofluorescence. Results showed that the level of a-SMA, collagen-I and fibronectin were notable with TGF-β1 stimulation. However, HSYA treatment obviously down regulated a-SMA and collagen-I protein expression, and slight down regulation of fibronectin (Fig 5A–5C).We further evaluated the expression of fibronectin and collagen-I via Western blot. Results showed that both fibronectin and collagen-I protein were higher in TGF-β1-stimulated group than in control group. HSYA treatment significantly ameliorated TGF-β1-induced increase of fibronectin and collagen-I expression (Fig 5D and 5E). These results indicated that HSYA could prevent the expression of the fibroblast marker and ECM in HK-2 cells, thus slow down the progression of TGF-β1-induced EMT.


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 myofibroblasts and accumulation of extracellular matrix in HK-2 cells.Immunofluorescence staining measured the expression levels of a-SMA (A), collagen-I (B) and fibronectin (C). Western blot analysis of FN and collagen-I protein levels (D). Semi-quantitative data from densitometric analysis of FN and collagen-I are presented as relative ratio of each protein to β-actin (E). 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.g005: Effect of HSYA on TGF-β1-induced myofibroblasts and accumulation of extracellular matrix in HK-2 cells.Immunofluorescence staining measured the expression levels of a-SMA (A), collagen-I (B) and fibronectin (C). Western blot analysis of FN and collagen-I protein levels (D). Semi-quantitative data from densitometric analysis of FN and collagen-I are presented as relative ratio of each protein to β-actin (E). 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: Due to the fact that the progression of EMT paves the way for extracellular matrix (ECM) deposition and finally deteriorate renal fibrosis, we detected the expression of a-SMA, collagen-I and fibronectin by Immunofluorescence. Results showed that the level of a-SMA, collagen-I and fibronectin were notable with TGF-β1 stimulation. However, HSYA treatment obviously down regulated a-SMA and collagen-I protein expression, and slight down regulation of fibronectin (Fig 5A–5C).We further evaluated the expression of fibronectin and collagen-I via Western blot. Results showed that both fibronectin and collagen-I protein were higher in TGF-β1-stimulated group than in control group. HSYA treatment significantly ameliorated TGF-β1-induced increase of fibronectin and collagen-I expression (Fig 5D and 5E). These results indicated that HSYA could prevent the expression of the fibroblast marker and ECM in HK-2 cells, thus slow down the progression of TGF-β1-induced EMT.

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