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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 attenuated liver injury and fibrogenesis in the TAA rat model. Representative liver sections were obtained from groups as follows: control, TAA and osthole-treated TAA rats. Liver sections were analyzed with a H&E-, b Picrosirius red- and c immunostaining of α-SMA. d Fibrosis scores were assessed by a pathologist in a blind fashion. e Quantification of collagen-positive area was performed using Metamorph software. f α-SMA protein in liver tissues detected by Western blotting analysis. g The expressions of α-sma and procollagen I, icam-1, mmp2, mmp9 and mmp13 transcripts in livers measured by qRT-PCR. h ELISA of TIMP-1, ICAM-1, CXCL7, CD62L, VEGF, and CX3CL1 in plasma of various groups. Data are shown as mean ± SD of 8 rats in each group. *p < 0.05; **p < 0.01, compared with other groups
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Fig2: Osthole attenuated liver injury and fibrogenesis in the TAA rat model. Representative liver sections were obtained from groups as follows: control, TAA and osthole-treated TAA rats. Liver sections were analyzed with a H&E-, b Picrosirius red- and c immunostaining of α-SMA. d Fibrosis scores were assessed by a pathologist in a blind fashion. e Quantification of collagen-positive area was performed using Metamorph software. f α-SMA protein in liver tissues detected by Western blotting analysis. g The expressions of α-sma and procollagen I, icam-1, mmp2, mmp9 and mmp13 transcripts in livers measured by qRT-PCR. h ELISA of TIMP-1, ICAM-1, CXCL7, CD62L, VEGF, and CX3CL1 in plasma of various groups. Data are shown as mean ± SD of 8 rats in each group. *p < 0.05; **p < 0.01, compared with other groups

Mentions: The function of osthole on TAA-induced hepatic injury and fibrosis were then evaluated based on the representative H&E- and Picrosirius red-stained liver sections. We observed changes in histological examination of livers from TAA rats compared with control rats, such as damage of hepatocytes and progressive increase and expansion of fibrous septa. Histological analysis revealed the suppressive effects of osthole in TAA-induced hepatic injury and fibrosis (Fig. 2a). Picrosirius red stain showed collagen fibres deposited in TAA-treated livers significantly compared with control rats (Fig. 2b). In addition, decreased α-SMA-positive cells were found in the fibrous septa portal tracts and sinusoids of the TAA + osthole-treated livers by immunohistochemical staining compared with TAA-treated livers (Fig. 2c). Consistently, the fibrosis score in TAA rats (0.72 ± 0.10) was significantly higher than that in control rats (0.14 ± 0.09), and osthole treatment (0.21 ± 0.14) significantly ameliorated hepatic fibrosis in TAA rats (Fig. 2d). The results showed that TAA intoxication led to a significant increase of collagen-positive area in the rat liver, which was attenuated by osthole treatment (Fig. 2e). We further confirmed that osthole treatment significantly attenuated TAA-induced α-SMA protein expression by Western blotting analysis (Fig. 2f). TAA-induced fibrosis-related gene expression, such as α-sma and procollagen I, was significantly reduced by osthole administration. There were also obvious decreases of mRNA levels in ECM synthesis-related genes, including icam-1, mmp2, mmp9 and mmp13 (Fig. 2g). To investigate whether osthole treatment affects the secretion of TAA-induced ECM-related cytokines and chemokines, we measured the production of distinct ECM-formation mediators by cytokine array. Results illustrated that osthole treatment exhibited a significant reduction of ICAM-1, CD62L, VEGF, and CX3CL1 levels. TIMP-1 and CXCL7 levels in the TAA + osthole group tended to be lower than those in the TAA group, but did not reach statistic significance (Fig. 2h). The data suggested that osthole treatment alleviated TAA-caused liver fibrosis and ECM synthesis.Fig. 2


Osthole ameliorates hepatic fibrosis and inhibits hepatic stellate cell activation.

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

Osthole attenuated liver injury and fibrogenesis in the TAA rat model. Representative liver sections were obtained from groups as follows: control, TAA and osthole-treated TAA rats. Liver sections were analyzed with a H&E-, b Picrosirius red- and c immunostaining of α-SMA. d Fibrosis scores were assessed by a pathologist in a blind fashion. e Quantification of collagen-positive area was performed using Metamorph software. f α-SMA protein in liver tissues detected by Western blotting analysis. g The expressions of α-sma and procollagen I, icam-1, mmp2, mmp9 and mmp13 transcripts in livers measured by qRT-PCR. h ELISA of TIMP-1, ICAM-1, CXCL7, CD62L, VEGF, and CX3CL1 in plasma of various groups. Data are shown as mean ± SD of 8 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

Fig2: Osthole attenuated liver injury and fibrogenesis in the TAA rat model. Representative liver sections were obtained from groups as follows: control, TAA and osthole-treated TAA rats. Liver sections were analyzed with a H&E-, b Picrosirius red- and c immunostaining of α-SMA. d Fibrosis scores were assessed by a pathologist in a blind fashion. e Quantification of collagen-positive area was performed using Metamorph software. f α-SMA protein in liver tissues detected by Western blotting analysis. g The expressions of α-sma and procollagen I, icam-1, mmp2, mmp9 and mmp13 transcripts in livers measured by qRT-PCR. h ELISA of TIMP-1, ICAM-1, CXCL7, CD62L, VEGF, and CX3CL1 in plasma of various groups. Data are shown as mean ± SD of 8 rats in each group. *p < 0.05; **p < 0.01, compared with other groups
Mentions: The function of osthole on TAA-induced hepatic injury and fibrosis were then evaluated based on the representative H&E- and Picrosirius red-stained liver sections. We observed changes in histological examination of livers from TAA rats compared with control rats, such as damage of hepatocytes and progressive increase and expansion of fibrous septa. Histological analysis revealed the suppressive effects of osthole in TAA-induced hepatic injury and fibrosis (Fig. 2a). Picrosirius red stain showed collagen fibres deposited in TAA-treated livers significantly compared with control rats (Fig. 2b). In addition, decreased α-SMA-positive cells were found in the fibrous septa portal tracts and sinusoids of the TAA + osthole-treated livers by immunohistochemical staining compared with TAA-treated livers (Fig. 2c). Consistently, the fibrosis score in TAA rats (0.72 ± 0.10) was significantly higher than that in control rats (0.14 ± 0.09), and osthole treatment (0.21 ± 0.14) significantly ameliorated hepatic fibrosis in TAA rats (Fig. 2d). The results showed that TAA intoxication led to a significant increase of collagen-positive area in the rat liver, which was attenuated by osthole treatment (Fig. 2e). We further confirmed that osthole treatment significantly attenuated TAA-induced α-SMA protein expression by Western blotting analysis (Fig. 2f). TAA-induced fibrosis-related gene expression, such as α-sma and procollagen I, was significantly reduced by osthole administration. There were also obvious decreases of mRNA levels in ECM synthesis-related genes, including icam-1, mmp2, mmp9 and mmp13 (Fig. 2g). To investigate whether osthole treatment affects the secretion of TAA-induced ECM-related cytokines and chemokines, we measured the production of distinct ECM-formation mediators by cytokine array. Results illustrated that osthole treatment exhibited a significant reduction of ICAM-1, CD62L, VEGF, and CX3CL1 levels. TIMP-1 and CXCL7 levels in the TAA + osthole group tended to be lower than those in the TAA group, but did not reach statistic significance (Fig. 2h). The data suggested that osthole treatment alleviated TAA-caused liver fibrosis and ECM synthesis.Fig. 2

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