Limits...
Moniliformediquinone as a potential therapeutic agent, inactivation of hepatic stellate cell and inhibition of liver fibrosis in vivo

View Article: PubMed Central - PubMed

ABSTRACT

Background: Moniliformediquinone (MFD), a phenanthradiquinone in Dendrobium moniliforme, was synthesized in our laboratory. Beyond its in vitro inhibitory effects on cancer cells, other biological activity of MFD is unknown. The purpose of the present study was to investigate the effects of MFD on hepatic fibrogenesis in vitro and in vivo.

Methods: Hepatic stellate HSC-T6 was cultured. Cell viability assay and western blot analyses were performed. Male ICR mice were evaluated on CCl4-induced hepatotoxicity using both histological examination and immunohistochemical staining.

Results: First, in vitro study showed that the synthesized MFD effectively attenuated the expression of transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), α-smooth muscle actin (α-SMA), and type I collagen (COL-1), which modulated the hepatic fibrogenesis. Furthermore, MFD reduced the phosphorylation of p65 NFκB in HSC-T6 cells. In vivo, liver fibrosis was induced by CCl4 twice a week for 10 weeks in mice. The administration of the MFD was started after 1 week of CCl4 thrice-weekly; the MFD significantly reduced plasma aspartate transaminase (AST) and lactose dehydrogenase (LDH) as well as hepatic hydroxy-proline, α-SMA, and COL-1 expression in CCl4-treated mice. Pathological analysis showed that the MFD alleviated CCl4-induced hepatic inflammation, necrosis and fibrosis. These results suggest that MFD possesses therapeutic potential for liver fibrosis.

Conclusions: The synthesized MFD exhibits anti-fibrotic potential by inactivation of HSCs in vitro and decreases mouse hepatic fibrosis in vivo. Further investigation into their clinical therapeutic potential is required.

No MeSH data available.


Related in: MedlinePlus

Effect of MFD on the expression of activated HSC-T6 markers and fibrosis-related proteins. HSC-T6 cells were treated with various concentrations of MFD for 48 h; then total cell lysates were tested for (a) TGF-β1, CTGF, α-SMA, COL-1 and p-p65 NFκB by western blot analysis. b HSC-T6 cells were treated with TGF-β1 (2 ng/mL) alone or in combination with different concentrations of MFD for 48 h; then total cell lysates were analyzed by western blot analysis against anti-CTGF, anti-α-SMA, anti-COL-1, and anti-β-actin as an internal control. Quantification of band intensity relative to α-tubulin is shown below the western blot. The quantitative data were presented as the mean of three repeats from one independent experiment. The data were presented as mean ± SD of three independent experiments. #P < 0.05 versus control. *P < 0.05, versus TGF-β1-treated alone
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC5017031&req=5

Fig2: Effect of MFD on the expression of activated HSC-T6 markers and fibrosis-related proteins. HSC-T6 cells were treated with various concentrations of MFD for 48 h; then total cell lysates were tested for (a) TGF-β1, CTGF, α-SMA, COL-1 and p-p65 NFκB by western blot analysis. b HSC-T6 cells were treated with TGF-β1 (2 ng/mL) alone or in combination with different concentrations of MFD for 48 h; then total cell lysates were analyzed by western blot analysis against anti-CTGF, anti-α-SMA, anti-COL-1, and anti-β-actin as an internal control. Quantification of band intensity relative to α-tubulin is shown below the western blot. The quantitative data were presented as the mean of three repeats from one independent experiment. The data were presented as mean ± SD of three independent experiments. #P < 0.05 versus control. *P < 0.05, versus TGF-β1-treated alone

Mentions: Transforming growth factor-β1 (TGF-β1) is a major fibrotic growth factor in liver fibrosis [9]. CTGF is selectively induced by TGF-β in a fibroblastic cell type and plays a key role in the overproduction of ECM in activated HSCs. Therefore, we determined the levels of TGF-β1 and CTGF after treatment with MFD in HSC-T6 cells using western blot analysis. The results showed that MFD significantly decreased the protein level of TGF-β1 and CTGF (Fig. 2a). In addition, the most striking biological consequences of activated HSCs are the marked accumulation of α-SMA and the dramatic increase in collagen depositions. MFD reduced the levels of α-SMA and COL-1 after treatment with MFD in HSC-T6 cells as compared with the control (Fig. 2a). Furthermore, MFD inhibited the phosphorylation of p65 NFkB (Fig. 2a), which has been suggested to be associated with the regulation of TGF-β1 expression [22]. To confirm the important role of TGF-β1 affected by MFD, TGF-β1 was added to promote the expression of CTGF, α-SMA, and COL-1. As shown in Fig. 2b, MFD reduced the expression of CTGF, α-SMA, and COL-1, which were mediated by TGF-β1.Fig. 2


Moniliformediquinone as a potential therapeutic agent, inactivation of hepatic stellate cell and inhibition of liver fibrosis in vivo
Effect of MFD on the expression of activated HSC-T6 markers and fibrosis-related proteins. HSC-T6 cells were treated with various concentrations of MFD for 48 h; then total cell lysates were tested for (a) TGF-β1, CTGF, α-SMA, COL-1 and p-p65 NFκB by western blot analysis. b HSC-T6 cells were treated with TGF-β1 (2 ng/mL) alone or in combination with different concentrations of MFD for 48 h; then total cell lysates were analyzed by western blot analysis against anti-CTGF, anti-α-SMA, anti-COL-1, and anti-β-actin as an internal control. Quantification of band intensity relative to α-tubulin is shown below the western blot. The quantitative data were presented as the mean of three repeats from one independent experiment. The data were presented as mean ± SD of three independent experiments. #P < 0.05 versus control. *P < 0.05, versus TGF-β1-treated alone
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5017031&req=5

Fig2: Effect of MFD on the expression of activated HSC-T6 markers and fibrosis-related proteins. HSC-T6 cells were treated with various concentrations of MFD for 48 h; then total cell lysates were tested for (a) TGF-β1, CTGF, α-SMA, COL-1 and p-p65 NFκB by western blot analysis. b HSC-T6 cells were treated with TGF-β1 (2 ng/mL) alone or in combination with different concentrations of MFD for 48 h; then total cell lysates were analyzed by western blot analysis against anti-CTGF, anti-α-SMA, anti-COL-1, and anti-β-actin as an internal control. Quantification of band intensity relative to α-tubulin is shown below the western blot. The quantitative data were presented as the mean of three repeats from one independent experiment. The data were presented as mean ± SD of three independent experiments. #P < 0.05 versus control. *P < 0.05, versus TGF-β1-treated alone
Mentions: Transforming growth factor-β1 (TGF-β1) is a major fibrotic growth factor in liver fibrosis [9]. CTGF is selectively induced by TGF-β in a fibroblastic cell type and plays a key role in the overproduction of ECM in activated HSCs. Therefore, we determined the levels of TGF-β1 and CTGF after treatment with MFD in HSC-T6 cells using western blot analysis. The results showed that MFD significantly decreased the protein level of TGF-β1 and CTGF (Fig. 2a). In addition, the most striking biological consequences of activated HSCs are the marked accumulation of α-SMA and the dramatic increase in collagen depositions. MFD reduced the levels of α-SMA and COL-1 after treatment with MFD in HSC-T6 cells as compared with the control (Fig. 2a). Furthermore, MFD inhibited the phosphorylation of p65 NFkB (Fig. 2a), which has been suggested to be associated with the regulation of TGF-β1 expression [22]. To confirm the important role of TGF-β1 affected by MFD, TGF-β1 was added to promote the expression of CTGF, α-SMA, and COL-1. As shown in Fig. 2b, MFD reduced the expression of CTGF, α-SMA, and COL-1, which were mediated by TGF-β1.Fig. 2

View Article: PubMed Central - PubMed

ABSTRACT

Background: Moniliformediquinone (MFD), a phenanthradiquinone in Dendrobium moniliforme, was synthesized in our laboratory. Beyond its in vitro inhibitory effects on cancer cells, other biological activity of MFD is unknown. The purpose of the present study was to investigate the effects of MFD on hepatic fibrogenesis in vitro and in vivo.

Methods: Hepatic stellate HSC-T6 was cultured. Cell viability assay and western blot analyses were performed. Male ICR mice were evaluated on CCl4-induced hepatotoxicity using both histological examination and immunohistochemical staining.

Results: First, in vitro study showed that the synthesized MFD effectively attenuated the expression of transforming growth factor-&beta;1 (TGF-&beta;1), connective tissue growth factor (CTGF), &alpha;-smooth muscle actin (&alpha;-SMA), and type I collagen (COL-1), which modulated the hepatic fibrogenesis. Furthermore, MFD reduced the phosphorylation of p65 NF&kappa;B in HSC-T6 cells. In vivo, liver fibrosis was induced by CCl4 twice a week for 10&nbsp;weeks in mice. The administration of the MFD was started after 1&nbsp;week of CCl4 thrice-weekly; the MFD significantly reduced plasma aspartate transaminase (AST) and lactose dehydrogenase (LDH) as well as hepatic hydroxy-proline, &alpha;-SMA, and COL-1 expression in CCl4-treated mice. Pathological analysis showed that the MFD alleviated CCl4-induced hepatic inflammation, necrosis and fibrosis. These results suggest that MFD possesses therapeutic potential for liver fibrosis.

Conclusions: The synthesized MFD exhibits anti-fibrotic potential by inactivation of HSCs in vitro and decreases mouse hepatic fibrosis in vivo. Further investigation into their clinical therapeutic potential is required.

No MeSH data available.


Related in: MedlinePlus