Limits...
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 and SIS3 onthe expression of α-SMA and E-cadherin in HK-2 cells.HK-2 cells treated with HSYA and SIS3for 48 h after stimulated with TGF-β1 (5 ng/ml). The a-SMA and E-cadherin expression level were detected by western blot. β-actin was used as a loading control. Semi-quantitative data from densitometric analysis of a-SMA and E-cadherin are presented as mean±SD, n = 3. #P<0.05 compared with control group; *P<0.05,**P<0.01 compared with TGF-β1-stimulated group.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4835075&req=5

pone.0153409.g009: Effect of HSYA and SIS3 onthe expression of α-SMA and E-cadherin in HK-2 cells.HK-2 cells treated with HSYA and SIS3for 48 h after stimulated with TGF-β1 (5 ng/ml). The a-SMA and E-cadherin expression level were detected by western blot. β-actin was used as a loading control. Semi-quantitative data from densitometric analysis of a-SMA and E-cadherin are presented as mean±SD, n = 3. #P<0.05 compared with control group; *P<0.05,**P<0.01 compared with TGF-β1-stimulated group.

Mentions: E-cadherin (the epithelial cell marker) and α-SMA (a specific myofibroblast marker) are usual biomarker of EMT. To further demonstrate the role of HSYA in the EMT process through TGF-β1/smads pathway, we treated HK-2withHSYA and SIS3 (5μM) after stimulated by human recombinant TGF-β1 (5ng/ml)and then to measure the expression of a-SMA and E-cadherin (Fig 9). The results showed that the expression level of a-SMA was reduced and level of E-cadherin was increased in treatment group of HSYA and SIS3, indicating that TGF-β1–induced EMT was reversed by inhibiting the smad3 signaling pathway (S6 Table). This suggested that activation of the TGF-β1/smad3 pathway appears to play a prominent role in the antifibrotic effect of HSYA.


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 and SIS3 onthe expression of α-SMA and E-cadherin in HK-2 cells.HK-2 cells treated with HSYA and SIS3for 48 h after stimulated with TGF-β1 (5 ng/ml). The a-SMA and E-cadherin expression level were detected by western blot. β-actin was used as a loading control. Semi-quantitative data from densitometric analysis of a-SMA and E-cadherin are presented as mean±SD, n = 3. #P<0.05 compared with control group; *P<0.05,**P<0.01 compared with TGF-β1-stimulated group.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153409.g009: Effect of HSYA and SIS3 onthe expression of α-SMA and E-cadherin in HK-2 cells.HK-2 cells treated with HSYA and SIS3for 48 h after stimulated with TGF-β1 (5 ng/ml). The a-SMA and E-cadherin expression level were detected by western blot. β-actin was used as a loading control. Semi-quantitative data from densitometric analysis of a-SMA and E-cadherin are presented as mean±SD, n = 3. #P<0.05 compared with control group; *P<0.05,**P<0.01 compared with TGF-β1-stimulated group.
Mentions: E-cadherin (the epithelial cell marker) and α-SMA (a specific myofibroblast marker) are usual biomarker of EMT. To further demonstrate the role of HSYA in the EMT process through TGF-β1/smads pathway, we treated HK-2withHSYA and SIS3 (5μM) after stimulated by human recombinant TGF-β1 (5ng/ml)and then to measure the expression of a-SMA and E-cadherin (Fig 9). The results showed that the expression level of a-SMA was reduced and level of E-cadherin was increased in treatment group of HSYA and SIS3, indicating that TGF-β1–induced EMT was reversed by inhibiting the smad3 signaling pathway (S6 Table). This suggested that activation of the TGF-β1/smad3 pathway appears to play a prominent role in the antifibrotic effect of HSYA.

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