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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.


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Effect of MFD on the liver fibrosis-related protein expression in CCl4-treated mouse liver. Histological examination of liver were revealed as indicated by immunohistochemical staining of TUNEL. Hydroxy-proline, α-SMA and COL-1 were evaluated by immunohistochemical staining. Representative liver sections stained as oil infusion control group (I); Mice with MFD treatment (II); Mice with CCl4 injection (III); MFD treatment (0.1 and 0.5 mg/kg) (IV, V); Magnification, ×200. The cells were counted from 10 fields (×200 magnification) of each liver sample. The results from statistical analysis are the means of cells and were calculated per microscope field from eight animals per group. Data are expressed as mean ± SD of independent experiments. *P < 0.05, as compared to the CCl4-treated group
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Fig4: Effect of MFD on the liver fibrosis-related protein expression in CCl4-treated mouse liver. Histological examination of liver were revealed as indicated by immunohistochemical staining of TUNEL. Hydroxy-proline, α-SMA and COL-1 were evaluated by immunohistochemical staining. Representative liver sections stained as oil infusion control group (I); Mice with MFD treatment (II); Mice with CCl4 injection (III); MFD treatment (0.1 and 0.5 mg/kg) (IV, V); Magnification, ×200. The cells were counted from 10 fields (×200 magnification) of each liver sample. The results from statistical analysis are the means of cells and were calculated per microscope field from eight animals per group. Data are expressed as mean ± SD of independent experiments. *P < 0.05, as compared to the CCl4-treated group

Mentions: The TUNEL assay results are shown in Fig. 4. Global immunoreactivity to apoptosis was localized primarily within liver tissue in CCl4 mouse. Quantitative examination of hepatocyte pathology showed that the number of normal hepatocytes present in the CCl4 treatment group was lower than the number of normal hepatocytes present in the control groups (control group = 5 ± 1; CCl4 treatment group = 65 ± 3; 0.1 mg/kg MFD treated CCl4 = 30 ± 3; 0.5 mg/kg MFD treated CCl4 = 15 ± 3, P < 0.05). We employed immunohistochemistry to examine liver hydroxy-proline protein expression in the hepatocyte of the untreated control and MFD alone group, the CCl4 group and the two CCl4 + MFD groups, which acts as an index of oxidative damage in hepatic fibrosis, was measured in order to investigate the therapeutic effect of MFD. A significant reduction in hydroxy-proline was found in the CCl4 + MFD groups compared to the CCl4 group. Significantly decreased expression of α-SMA, and COL-1 proteins were found in the MFD groups compared to the CCl4 group, P < 0.05 (Fig. 4).Fig. 4


Moniliformediquinone as a potential therapeutic agent, inactivation of hepatic stellate cell and inhibition of liver fibrosis in vivo
Effect of MFD on the liver fibrosis-related protein expression in CCl4-treated mouse liver. Histological examination of liver were revealed as indicated by immunohistochemical staining of TUNEL. Hydroxy-proline, α-SMA and COL-1 were evaluated by immunohistochemical staining. Representative liver sections stained as oil infusion control group (I); Mice with MFD treatment (II); Mice with CCl4 injection (III); MFD treatment (0.1 and 0.5 mg/kg) (IV, V); Magnification, ×200. The cells were counted from 10 fields (×200 magnification) of each liver sample. The results from statistical analysis are the means of cells and were calculated per microscope field from eight animals per group. Data are expressed as mean ± SD of independent experiments. *P < 0.05, as compared to the CCl4-treated group
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5017031&req=5

Fig4: Effect of MFD on the liver fibrosis-related protein expression in CCl4-treated mouse liver. Histological examination of liver were revealed as indicated by immunohistochemical staining of TUNEL. Hydroxy-proline, α-SMA and COL-1 were evaluated by immunohistochemical staining. Representative liver sections stained as oil infusion control group (I); Mice with MFD treatment (II); Mice with CCl4 injection (III); MFD treatment (0.1 and 0.5 mg/kg) (IV, V); Magnification, ×200. The cells were counted from 10 fields (×200 magnification) of each liver sample. The results from statistical analysis are the means of cells and were calculated per microscope field from eight animals per group. Data are expressed as mean ± SD of independent experiments. *P < 0.05, as compared to the CCl4-treated group
Mentions: The TUNEL assay results are shown in Fig. 4. Global immunoreactivity to apoptosis was localized primarily within liver tissue in CCl4 mouse. Quantitative examination of hepatocyte pathology showed that the number of normal hepatocytes present in the CCl4 treatment group was lower than the number of normal hepatocytes present in the control groups (control group = 5 ± 1; CCl4 treatment group = 65 ± 3; 0.1 mg/kg MFD treated CCl4 = 30 ± 3; 0.5 mg/kg MFD treated CCl4 = 15 ± 3, P < 0.05). We employed immunohistochemistry to examine liver hydroxy-proline protein expression in the hepatocyte of the untreated control and MFD alone group, the CCl4 group and the two CCl4 + MFD groups, which acts as an index of oxidative damage in hepatic fibrosis, was measured in order to investigate the therapeutic effect of MFD. A significant reduction in hydroxy-proline was found in the CCl4 + MFD groups compared to the CCl4 group. Significantly decreased expression of α-SMA, and COL-1 proteins were found in the MFD groups compared to the CCl4 group, P < 0.05 (Fig. 4).Fig. 4

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