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Potent effects of dioscin against liver fibrosis.

Zhang X, Han X, Yin L, Xu L, Qi Y, Xu Y, Sun H, Lin Y, Liu K, Peng J - Sci Rep (2015)

Bottom Line: Dioscin effectively inhibited the cell viabilities of HSC-T6, LX-2 and primary rat hepatic stellate cells (HSCs), but not hepatocytes.Furthermore, dioscin markedly increased peroxisome proliferator activated receptor-γ (PPAR-γ) expression and significantly reduced a-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), collagen α1 (I) (COL1A1) and collagen α1 (III) (COL3A1) levels in vitro.Dioscin facilitated matrix degradation, and exhibited hepatoprotective effects through the attenuation of oxidative stress and inflammation, in addition to exerting anti-fibrotic effects through the modulation of the TGF-β1/Smad, Wnt/β-catenin, mitogen-activated protein kinase (MAPK) and mitochondrial signaling pathways, which triggered the senescence of activated HSCs.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian 116044, China.

ABSTRACT
We previously reported the promising effects of dioscin against liver injury, but its effect on liver fibrosis remains unknown. The present work investigated the activities of dioscin against liver fibrosis and the underlying molecular mechanisms. Dioscin effectively inhibited the cell viabilities of HSC-T6, LX-2 and primary rat hepatic stellate cells (HSCs), but not hepatocytes. Furthermore, dioscin markedly increased peroxisome proliferator activated receptor-γ (PPAR-γ) expression and significantly reduced a-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), collagen α1 (I) (COL1A1) and collagen α1 (III) (COL3A1) levels in vitro. Notably, dioscin inhibited HSCs activation and induced apoptosis in activated HSCs. In vivo, dioscin significantly improved body weight and hydroxylproline, laminin, α-SMA, TGF-β1, COL1A1 and COL3A1 levels, which were confirmed by histopathological assays. Dioscin facilitated matrix degradation, and exhibited hepatoprotective effects through the attenuation of oxidative stress and inflammation, in addition to exerting anti-fibrotic effects through the modulation of the TGF-β1/Smad, Wnt/β-catenin, mitogen-activated protein kinase (MAPK) and mitochondrial signaling pathways, which triggered the senescence of activated HSCs. In conclusion, dioscin exhibited potent effects against liver fibrosis through the modulation of multiple targets and signaling pathways and should be developed as a novel candidate for the treatment of liver fibrosis in the future.

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Effects of dioscin on HSCs activation of HSC-T6, LX2 and primary HSCs.(A) Impacts of dioscin on the cell viabilities of HSC-T6, LX2, primary rat HSCs. After incubating for 24 14h, the cells were treated with various concentrations of dioscin (1.25, 2.5 and 5.0 14μg/ml) or 0.1% DMSO as a negative control for 12, 24 and 48 14h, then the cell viability was evaluated by MTT assay. (B) Effects of dioscin on PPAR-γ protein expression in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h. Total protein samples extracted from control and dioscin-treated HSCs were analyzed by western blotting assay (The cropped gels are used and full-length gels are presented in Supplemental Figure S9). (C) Effects of dioscin on the mRNA levels of TGF-β1, α-SMA, COL1A1 and COL3A1 in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h, and then total RNA samples were extracted from control and dioscin-treated HSCs, which were measured by real-time PCR assay. (D) Effects of dioscin on the levels of α-SMA in HSC-T6, LX2 and primary rat HSCs based on immunofluorescence assay (10000 × magnification). After treatment with various concentrations of dioscin for 12, 24 and 48 14h, the expression of α-SMA was detected by immunofluorescence, and DAPI was used to visualize the nucleus. Data are presented as the mean ± SD (n ≥ 3). *p < 0.05 and **p < 0.01 compared with the control group.
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f1: Effects of dioscin on HSCs activation of HSC-T6, LX2 and primary HSCs.(A) Impacts of dioscin on the cell viabilities of HSC-T6, LX2, primary rat HSCs. After incubating for 24 14h, the cells were treated with various concentrations of dioscin (1.25, 2.5 and 5.0 14μg/ml) or 0.1% DMSO as a negative control for 12, 24 and 48 14h, then the cell viability was evaluated by MTT assay. (B) Effects of dioscin on PPAR-γ protein expression in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h. Total protein samples extracted from control and dioscin-treated HSCs were analyzed by western blotting assay (The cropped gels are used and full-length gels are presented in Supplemental Figure S9). (C) Effects of dioscin on the mRNA levels of TGF-β1, α-SMA, COL1A1 and COL3A1 in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h, and then total RNA samples were extracted from control and dioscin-treated HSCs, which were measured by real-time PCR assay. (D) Effects of dioscin on the levels of α-SMA in HSC-T6, LX2 and primary rat HSCs based on immunofluorescence assay (10000 × magnification). After treatment with various concentrations of dioscin for 12, 24 and 48 14h, the expression of α-SMA was detected by immunofluorescence, and DAPI was used to visualize the nucleus. Data are presented as the mean ± SD (n ≥ 3). *p < 0.05 and **p < 0.01 compared with the control group.

Mentions: Figure 1A demonstrates that dioscin significantly inhibited the viability of HSC-T6, LX2 and primary activated HSCs with dose- and time-dependent manners. Dioscin at the concentration of 5 14μg/ml for 48 14h significantly decreased the cell viabilities to 49.84%, 40.52% and 45.26%, respectively. Notably, dioscin at concentrations of 1.25, 2.5 and 5.0 14μg/ml for 12, 24 and 48 14h did not obviously reduce the viability of primary cultured hepatocytes, suggesting that the inhibition of cell viability induced dioscin was specific to HSCs.


Potent effects of dioscin against liver fibrosis.

Zhang X, Han X, Yin L, Xu L, Qi Y, Xu Y, Sun H, Lin Y, Liu K, Peng J - Sci Rep (2015)

Effects of dioscin on HSCs activation of HSC-T6, LX2 and primary HSCs.(A) Impacts of dioscin on the cell viabilities of HSC-T6, LX2, primary rat HSCs. After incubating for 24 14h, the cells were treated with various concentrations of dioscin (1.25, 2.5 and 5.0 14μg/ml) or 0.1% DMSO as a negative control for 12, 24 and 48 14h, then the cell viability was evaluated by MTT assay. (B) Effects of dioscin on PPAR-γ protein expression in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h. Total protein samples extracted from control and dioscin-treated HSCs were analyzed by western blotting assay (The cropped gels are used and full-length gels are presented in Supplemental Figure S9). (C) Effects of dioscin on the mRNA levels of TGF-β1, α-SMA, COL1A1 and COL3A1 in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h, and then total RNA samples were extracted from control and dioscin-treated HSCs, which were measured by real-time PCR assay. (D) Effects of dioscin on the levels of α-SMA in HSC-T6, LX2 and primary rat HSCs based on immunofluorescence assay (10000 × magnification). After treatment with various concentrations of dioscin for 12, 24 and 48 14h, the expression of α-SMA was detected by immunofluorescence, and DAPI was used to visualize the nucleus. Data are presented as the mean ± SD (n ≥ 3). *p < 0.05 and **p < 0.01 compared with the control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4389718&req=5

f1: Effects of dioscin on HSCs activation of HSC-T6, LX2 and primary HSCs.(A) Impacts of dioscin on the cell viabilities of HSC-T6, LX2, primary rat HSCs. After incubating for 24 14h, the cells were treated with various concentrations of dioscin (1.25, 2.5 and 5.0 14μg/ml) or 0.1% DMSO as a negative control for 12, 24 and 48 14h, then the cell viability was evaluated by MTT assay. (B) Effects of dioscin on PPAR-γ protein expression in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h. Total protein samples extracted from control and dioscin-treated HSCs were analyzed by western blotting assay (The cropped gels are used and full-length gels are presented in Supplemental Figure S9). (C) Effects of dioscin on the mRNA levels of TGF-β1, α-SMA, COL1A1 and COL3A1 in HSC-T6, LX2 and primary rat HSCs. The cells were treated with various concentrations of dioscin for 12, 24 and 48 14h, and then total RNA samples were extracted from control and dioscin-treated HSCs, which were measured by real-time PCR assay. (D) Effects of dioscin on the levels of α-SMA in HSC-T6, LX2 and primary rat HSCs based on immunofluorescence assay (10000 × magnification). After treatment with various concentrations of dioscin for 12, 24 and 48 14h, the expression of α-SMA was detected by immunofluorescence, and DAPI was used to visualize the nucleus. Data are presented as the mean ± SD (n ≥ 3). *p < 0.05 and **p < 0.01 compared with the control group.
Mentions: Figure 1A demonstrates that dioscin significantly inhibited the viability of HSC-T6, LX2 and primary activated HSCs with dose- and time-dependent manners. Dioscin at the concentration of 5 14μg/ml for 48 14h significantly decreased the cell viabilities to 49.84%, 40.52% and 45.26%, respectively. Notably, dioscin at concentrations of 1.25, 2.5 and 5.0 14μg/ml for 12, 24 and 48 14h did not obviously reduce the viability of primary cultured hepatocytes, suggesting that the inhibition of cell viability induced dioscin was specific to HSCs.

Bottom Line: Dioscin effectively inhibited the cell viabilities of HSC-T6, LX-2 and primary rat hepatic stellate cells (HSCs), but not hepatocytes.Furthermore, dioscin markedly increased peroxisome proliferator activated receptor-γ (PPAR-γ) expression and significantly reduced a-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), collagen α1 (I) (COL1A1) and collagen α1 (III) (COL3A1) levels in vitro.Dioscin facilitated matrix degradation, and exhibited hepatoprotective effects through the attenuation of oxidative stress and inflammation, in addition to exerting anti-fibrotic effects through the modulation of the TGF-β1/Smad, Wnt/β-catenin, mitogen-activated protein kinase (MAPK) and mitochondrial signaling pathways, which triggered the senescence of activated HSCs.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian 116044, China.

ABSTRACT
We previously reported the promising effects of dioscin against liver injury, but its effect on liver fibrosis remains unknown. The present work investigated the activities of dioscin against liver fibrosis and the underlying molecular mechanisms. Dioscin effectively inhibited the cell viabilities of HSC-T6, LX-2 and primary rat hepatic stellate cells (HSCs), but not hepatocytes. Furthermore, dioscin markedly increased peroxisome proliferator activated receptor-γ (PPAR-γ) expression and significantly reduced a-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), collagen α1 (I) (COL1A1) and collagen α1 (III) (COL3A1) levels in vitro. Notably, dioscin inhibited HSCs activation and induced apoptosis in activated HSCs. In vivo, dioscin significantly improved body weight and hydroxylproline, laminin, α-SMA, TGF-β1, COL1A1 and COL3A1 levels, which were confirmed by histopathological assays. Dioscin facilitated matrix degradation, and exhibited hepatoprotective effects through the attenuation of oxidative stress and inflammation, in addition to exerting anti-fibrotic effects through the modulation of the TGF-β1/Smad, Wnt/β-catenin, mitogen-activated protein kinase (MAPK) and mitochondrial signaling pathways, which triggered the senescence of activated HSCs. In conclusion, dioscin exhibited potent effects against liver fibrosis through the modulation of multiple targets and signaling pathways and should be developed as a novel candidate for the treatment of liver fibrosis in the future.

Show MeSH
Related in: MedlinePlus