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, 2012, 212530

Recovery of the Cell Cycle Inhibition in CCl(4)-Induced Cirrhosis by the Adenosine Derivative IFC-305

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Recovery of the Cell Cycle Inhibition in CCl(4)-Induced Cirrhosis by the Adenosine Derivative IFC-305

Victoria Chagoya de Sánchez et al. Int J Hepatol.

Abstract

Introduction. Cirrhosis is a chronic degenerative illness characterized by changes in normal liver architecture, failure of hepatic function, and impairment of proliferative activity. The aim of this study is to know how IFC-305 compound induces proliferation of the liver during reversion of cirrhosis. Methods. Once cirrhosis has been installed by CCl(4) treatment for 10 weeks in male Wistar rats, they were divided into four groups: two received saline and two received the compound; all were euthanized at 5 and 10 weeks of treatment. Liver homogenate, mitochondria, and nucleus were used to measure cyclins, CDKs, and cell cycle regulatory proteins PCNA, pRb, p53, E2F, p21, p27, HGF, liver ATP, and mitochondrial function. Results. Diminution and small changes were observed in the studied proteins in the cirrhotic animals without treatment. The IFC-305-treated rats showed a clear increase in most of the proteins studied mainly in PCNA and CDK6, and a marked increased in ATP and mitochondrial function. Discussion/Conclusion. IFC-305 induces a recovery of the cell cycle inhibition promoting recovery of DNA damage through the action of PCNA and p53. The increase in energy and preservation of mitochondrial function contribute to recovering the proliferative function.

Figures

Figure 1
Figure 1
Effect of IFC305 treatment on the liver PCNA protein expression in CCl4-induced cirrhosis in rats. (a) Protein expression of PCNA and the house keeping gen β-actin from liver nuclear extracts was determined by western blot analysis. A representative western blot image is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group; values were normalized to β-actin immunodetection. (b) Immunohistochemical analysis of PCNA expression. Hepatocyte (arrowhead) and nonparenchymal cell (arrow) are marked. The number of PCNA positive hepatocytes was quantified as described in Section 2. *Statistical difference (P < 0.01) when compared to C group. #Statistical difference (P < 0.01) when compared to their respective experimental group SS5 or SS10.
Figure 2
Figure 2
Effect of IFC305 treatment on cyclin D1, cyclin E, cyclin A, and cyclin B1 protein expression in CCl4-induced cirrhosis in rats. (a), (c), (d), and (e) Expression of the indicated proteins from liver nuclear extracts was determined by western blot analysis. A representative western blot image of each one is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group, values were normalized to β-actin immunodetection (image, Figure 1(a)). *Statistical difference (P < 0.05) when compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10. (b) Effect of IFC305 treatment on cyclin D1 mRNA expression in CCl4-induced cirrhosis in rats. RNA was isolated from liver and mRNA expression for cyclin D1 was analyzed by quantitative RT-PCR as described in Section 2. Arbitrary units were normalized with Arbp mRNA gene expression level. Data represent mean ± SEM from 3 rats/group. *Statistical difference (P < 0.05) compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10.
Figure 3
Figure 3
Effect of IFC305 treatment on CDK4, CDK6, p21, and p27 proteins expression in cirrhotic livers. (a), (b), (c), and (d) Expression of the indicated proteins from liver nuclear extracts was done by western blot analysis. A representative western blot image is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group; values were normalized to β-actin immunodetection (image, Figure 1(a)). *Statistical difference (P < 0.05) when compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10.
Figure 4
Figure 4
Effect of IFC305 treatment on phospho-Rb (Ser 795), E2F1, and DP1 proteins expression in cirrhotic livers. (a), (c), and (d) Expression of the indicated proteins from liver nuclear extracts was done by western blot analysis. A representative western blot image of each one is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group; values were normalized to β-actin immunodetection (image, Figure 1(a)). *Statistical difference (P < 0.05) when compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10. (b) Effect of IFC305 treatment on Rb mRNA expression in CCl4-induced cirrhosis in rats. RNA was isolated from liver and mRNA expression for Rb was analyzed by quantitative RT-PCR as described in Section 2. Arbitrary units were normalized with Arbp mRNA gene expression level. Data represent mean ± SEM from 3 rats/group. *Statistical difference (P < 0.05) compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10.
Figure 5
Figure 5
Effect of IFC305 treatment on p53 and MDM2 proteins expression in cirrhotic livers. (a) and (c) Expression of the indicated proteins from liver nuclear extracts was done by western blot analysis. A representative western blot image of each one is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group; values were normalized to β-actin immunodetection (image, Figure 1(a)). *Statistical difference (P < 0.05) when compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10. (b) Effect of IFC305 treatment on p53 mRNA expression in CCl4-induced cirrhosis in rats. RNA was isolated from liver and mRNA expression for Rb was analyzed by quantitative RT-PCR as described in Section 2. Arbitrary units were normalized with Arbp mRNA gene expression level. Data represent mean ± SEM from 3 rats/group. *Statistical difference (P < 0.05) when compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10.
Figure 6
Figure 6
Effect of IFC305 treatment on serum HGF, liver HGF, and cMet proteins expression in CCl4-induced cirrhosis in rats. (a), (b), and (c) Expression of the indicated proteins from serum or liver extracts was done by western blot analysis. A representative western blot image of each one is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group; values were normalized to β-actin immunodetection (image, Figure 1(a)). Values from serum (a) were not normalized. *Statistical difference (P < 0.05) when compared to C group. #Statistical difference (P < 0.05) when compared to their respective experimental group SS5 or SS10.
Figure 7
Figure 7
Effect of IFC305 treatment on adenosine receptors expression in CCl4-induced cirrhosis in rats. Expression of the indicated adenosine receptors from liver extracts was done by western blot analysis. A representative western blot image of each one is shown. The bar graph represents the densitometry analysis expressed as arbitrary units of the mean ± SEM from 3 rats/group; values were normalized to β-actin immunodetection (image, Figure 1(a)).

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References

    1. Bruck R, Hershkoviz R, Lider O, et al. Inhibition of experimentally-induced liver cirrhosis in rats by a nonpeptidic mimetic of the extracellular matrix-associated Arg-Gly-Asp epitope. Journal of Hepatology. 1996;24(6):731–738. - PubMed
    1. Tamayo RP. Is cirrhosis of the liver experimentally produced by CCl4 an adequate model of human cirrhosis? Hepatology. 1983;3(1):112–120. - PubMed
    1. Kokudo N, Kothary PC, Eckhauser FE, Raper SE. Transforming growth factor-α (TGF-α) improves hepatic DNA synthesis after hepatectomy in cirrhotic rats. Journal of Surgical Research. 1992;52(6):648–655. - PubMed
    1. Kato A, Bamba H, Shinohara M, et al. Relationship between expression of cyclin D1 and impaired liver regeneration observed in fibrotic or cirrhotic rats. Journal of Gastroenterology and Hepatology. 2005;20(8):1198–1205. - PubMed
    1. de Sánchez VC, Hernández-Luis F, Díaz-Muñoz M, Hernández-Muñoz R. Role of the Energy State of Liver Cell in Cirrosis Development and Treatment. Hauppauge, NY, USA: NOVA Science; 2011.
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