Limits...
Osthole ameliorates hepatic fibrosis and inhibits hepatic stellate cell activation.

Liu YW, Chiu YT, Fu SL, Huang YT - J. Biomed. Sci. (2015)

Bottom Line: Additionally, osthole reduced the expression of fibrosis-related genes significantly.Furthermore, osthole decreased TNF-α-triggered NF-κB activities significantly.In addition, osthole suppressed HSCs activation in vitro significantly.

View Article: PubMed Central - PubMed

Affiliation: Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, No. 155, Li-Nong Street, Sec. 2, Taipei, 11221, Taiwan. yaweiliu19850417@gmail.com.

ABSTRACT

Background: Hepatic fibrosis is a dynamic process which ultimately leads to cirrhosis in almost patients with chronic hepatic injury. However, progressive fibrosis is a reversible scarring response. Activation of hepatic stellate cells (HSCs) is the prevailing process during hepatic fibrosis. Osthole is an active component majorly contained in the fruit of Cnidium monnieri (L.) Cusson. This present study investigated the therapeutic effects of osthole on rat liver fibrosis and HSC activation.

Results: We established the thioacetamide (TAA)-model of Sprague-Dawley (SD) rats to induce hepatic fibrosis. Rats were divided into three groups: control, TAA, and TAA + osthole (10 mg/kg). In vivo, osthole significantly reduced liver injury by diminishing levels of plasma AST and ALT, improving histological architecture, decreasing collagen and α-SMA accumulation, and improving hepatic fibrosis scores. Additionally, osthole reduced the expression of fibrosis-related genes significantly. Osthole also suppressed the production of fibrosis-related cytokines and chemokines. Moreover, nuclear translocation of p65 was significantly suppressed in osthole-treated liver. Osthole also ameliorated TAA-induced injury through reducing cellular oxidation. Osthole showed inhibitory effects in inflammation-related genes and chemokines production as well. In vitro, we assessed osthole effects in activated HSCs (HSC-T6 and LX-2). Osthole attenuated TGF-β1-induced migration and invasion in HSCs. Furthermore, osthole decreased TNF-α-triggered NF-κB activities significantly. Besides, osthole alleviated TGF-β1- or ET-1-induced HSCs contractility.

Conclusions: Our study demonstrated that osthole improved TAA-caused liver injury, fibrogenesis and inflammation in rats. In addition, osthole suppressed HSCs activation in vitro significantly.

No MeSH data available.


Related in: MedlinePlus

Osthole suppressed inflammation and oxidation in TAA-injected rat liver. a Nuclear fractions of rat liver from different treatment groups were analyzed by Western blotting analysis. For expression of p65, JunD and Nrf-2, PCNA expression served as loading control in nuclear protein. b GSH/GSSG ratio in the livers was measured by Glutathione kit. c Levels of 4-HNE in the livers were measured using 4-HNE ELISA kit. d Levels of MDA in the livers were measured using MDA ELISA kit e qRT-PCR showed the expressions of interleukin-1β, tnf-α and inos transcripts in rats. f ELISA of LIX, CXCL1, CCL20 and CCL5 in plasma. Data are shown as mean ± SD of 6 rats in each group. *p < 0.05; **p < 0.01, compared with other groups
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4522080&req=5

Fig3: Osthole suppressed inflammation and oxidation in TAA-injected rat liver. a Nuclear fractions of rat liver from different treatment groups were analyzed by Western blotting analysis. For expression of p65, JunD and Nrf-2, PCNA expression served as loading control in nuclear protein. b GSH/GSSG ratio in the livers was measured by Glutathione kit. c Levels of 4-HNE in the livers were measured using 4-HNE ELISA kit. d Levels of MDA in the livers were measured using MDA ELISA kit e qRT-PCR showed the expressions of interleukin-1β, tnf-α and inos transcripts in rats. f ELISA of LIX, CXCL1, CCL20 and CCL5 in plasma. Data are shown as mean ± SD of 6 rats in each group. *p < 0.05; **p < 0.01, compared with other groups

Mentions: In chronic liver diseases, inflammation is a major feature associated with fibrogenesis [21, 22]. We postulated that osthole treatment might attenuate TAA-induced injury by inhibiting liver inflammation. The H&E-stained liver sections in TAA injected rats showed portal triad inflammation and periportal inflammatory cell infiltration (Fig. 2a). We further prepared nuclear fractions to determine translocation of p65 and JunD. The p65 is the main subunit of NF-κB which can be triggered to translocate into the nucleus then activating transcription of various genes. JunD is a functional component of the activator protein-1 (AP-1) transcription factor complex known to activate AP-1-mediated transcription of genes in inflammatory response [23]. Data of nuclear fraction using Western blotting analysis indicated that there were more p65 and JunD nuclear translocation in TAA-injected livers than in the control group. Osthole treatment evidently reduced p65 translocation into nuclei in TAA-injected rats (Fig. 3a). The JunD translocation in osthole-treatment group tended to be lower than those in the TAA group. Nrf-2 plays a critical role in stress-inducible genes and also in cellular resistance to oxidants. TAA-promoted Nrf-2 translocation was profoundly down-regulated by osthole treatment, suggesting that oxidative stress in the liver might be counteracted by osthole (Fig. 3a). In Glutathione assay, we measured the levels of total GSH and oxidized GSSG, and then determined the level of GSH/GSSG ratio. The results showed that TAA treatment led to higher GSH/GSSG ratio, and osthole treatment restored GSH depletion in the liver (Fig. 3b). In addition, we found that osthole reduced the amount of TAA-caused lipid peroxidative products 4-HNE and MDA in the liver (Fig. 3c, d). To further explore the anti-inflammatory role of osthole treatment, the mRNA levels of interleukin-1β, tnf-α and inos genes in the rat liver were evaluated by qRT-PCR. The expression levels of these inflammation-related genes in the liver were increased in TAA-injected rats, which were obviously attenuated by osthole treatment (Fig. 3e). Additionally, levels of inflammation-related chemokines in the TAA group were significantly higher than those in control rats, but only CXCL1 levels showed significant reduction after osthole treatment. LIX and CCL20 levels in the TAA + osthole group tended to be lower than those in the TAA group, but did not reach statistic significance (Fig. 3f).Fig. 3


Osthole ameliorates hepatic fibrosis and inhibits hepatic stellate cell activation.

Liu YW, Chiu YT, Fu SL, Huang YT - J. Biomed. Sci. (2015)

Osthole suppressed inflammation and oxidation in TAA-injected rat liver. a Nuclear fractions of rat liver from different treatment groups were analyzed by Western blotting analysis. For expression of p65, JunD and Nrf-2, PCNA expression served as loading control in nuclear protein. b GSH/GSSG ratio in the livers was measured by Glutathione kit. c Levels of 4-HNE in the livers were measured using 4-HNE ELISA kit. d Levels of MDA in the livers were measured using MDA ELISA kit e qRT-PCR showed the expressions of interleukin-1β, tnf-α and inos transcripts in rats. f ELISA of LIX, CXCL1, CCL20 and CCL5 in plasma. Data are shown as mean ± SD of 6 rats in each group. *p < 0.05; **p < 0.01, compared with other groups
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4522080&req=5

Fig3: Osthole suppressed inflammation and oxidation in TAA-injected rat liver. a Nuclear fractions of rat liver from different treatment groups were analyzed by Western blotting analysis. For expression of p65, JunD and Nrf-2, PCNA expression served as loading control in nuclear protein. b GSH/GSSG ratio in the livers was measured by Glutathione kit. c Levels of 4-HNE in the livers were measured using 4-HNE ELISA kit. d Levels of MDA in the livers were measured using MDA ELISA kit e qRT-PCR showed the expressions of interleukin-1β, tnf-α and inos transcripts in rats. f ELISA of LIX, CXCL1, CCL20 and CCL5 in plasma. Data are shown as mean ± SD of 6 rats in each group. *p < 0.05; **p < 0.01, compared with other groups
Mentions: In chronic liver diseases, inflammation is a major feature associated with fibrogenesis [21, 22]. We postulated that osthole treatment might attenuate TAA-induced injury by inhibiting liver inflammation. The H&E-stained liver sections in TAA injected rats showed portal triad inflammation and periportal inflammatory cell infiltration (Fig. 2a). We further prepared nuclear fractions to determine translocation of p65 and JunD. The p65 is the main subunit of NF-κB which can be triggered to translocate into the nucleus then activating transcription of various genes. JunD is a functional component of the activator protein-1 (AP-1) transcription factor complex known to activate AP-1-mediated transcription of genes in inflammatory response [23]. Data of nuclear fraction using Western blotting analysis indicated that there were more p65 and JunD nuclear translocation in TAA-injected livers than in the control group. Osthole treatment evidently reduced p65 translocation into nuclei in TAA-injected rats (Fig. 3a). The JunD translocation in osthole-treatment group tended to be lower than those in the TAA group. Nrf-2 plays a critical role in stress-inducible genes and also in cellular resistance to oxidants. TAA-promoted Nrf-2 translocation was profoundly down-regulated by osthole treatment, suggesting that oxidative stress in the liver might be counteracted by osthole (Fig. 3a). In Glutathione assay, we measured the levels of total GSH and oxidized GSSG, and then determined the level of GSH/GSSG ratio. The results showed that TAA treatment led to higher GSH/GSSG ratio, and osthole treatment restored GSH depletion in the liver (Fig. 3b). In addition, we found that osthole reduced the amount of TAA-caused lipid peroxidative products 4-HNE and MDA in the liver (Fig. 3c, d). To further explore the anti-inflammatory role of osthole treatment, the mRNA levels of interleukin-1β, tnf-α and inos genes in the rat liver were evaluated by qRT-PCR. The expression levels of these inflammation-related genes in the liver were increased in TAA-injected rats, which were obviously attenuated by osthole treatment (Fig. 3e). Additionally, levels of inflammation-related chemokines in the TAA group were significantly higher than those in control rats, but only CXCL1 levels showed significant reduction after osthole treatment. LIX and CCL20 levels in the TAA + osthole group tended to be lower than those in the TAA group, but did not reach statistic significance (Fig. 3f).Fig. 3

Bottom Line: Additionally, osthole reduced the expression of fibrosis-related genes significantly.Furthermore, osthole decreased TNF-α-triggered NF-κB activities significantly.In addition, osthole suppressed HSCs activation in vitro significantly.

View Article: PubMed Central - PubMed

Affiliation: Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, No. 155, Li-Nong Street, Sec. 2, Taipei, 11221, Taiwan. yaweiliu19850417@gmail.com.

ABSTRACT

Background: Hepatic fibrosis is a dynamic process which ultimately leads to cirrhosis in almost patients with chronic hepatic injury. However, progressive fibrosis is a reversible scarring response. Activation of hepatic stellate cells (HSCs) is the prevailing process during hepatic fibrosis. Osthole is an active component majorly contained in the fruit of Cnidium monnieri (L.) Cusson. This present study investigated the therapeutic effects of osthole on rat liver fibrosis and HSC activation.

Results: We established the thioacetamide (TAA)-model of Sprague-Dawley (SD) rats to induce hepatic fibrosis. Rats were divided into three groups: control, TAA, and TAA + osthole (10 mg/kg). In vivo, osthole significantly reduced liver injury by diminishing levels of plasma AST and ALT, improving histological architecture, decreasing collagen and α-SMA accumulation, and improving hepatic fibrosis scores. Additionally, osthole reduced the expression of fibrosis-related genes significantly. Osthole also suppressed the production of fibrosis-related cytokines and chemokines. Moreover, nuclear translocation of p65 was significantly suppressed in osthole-treated liver. Osthole also ameliorated TAA-induced injury through reducing cellular oxidation. Osthole showed inhibitory effects in inflammation-related genes and chemokines production as well. In vitro, we assessed osthole effects in activated HSCs (HSC-T6 and LX-2). Osthole attenuated TGF-β1-induced migration and invasion in HSCs. Furthermore, osthole decreased TNF-α-triggered NF-κB activities significantly. Besides, osthole alleviated TGF-β1- or ET-1-induced HSCs contractility.

Conclusions: Our study demonstrated that osthole improved TAA-caused liver injury, fibrogenesis and inflammation in rats. In addition, osthole suppressed HSCs activation in vitro significantly.

No MeSH data available.


Related in: MedlinePlus