Limits...
Lack of CC chemokine ligand 2 differentially affects inflammation and fibrosis according to the genetic background in a murine model of steatohepatitis.

Galastri S, Zamara E, Milani S, Novo E, Provenzano A, Delogu W, Vizzutti F, Sutti S, Locatelli I, Navari N, Vivoli E, Caligiuri A, Pinzani M, Albano E, Parola M, Marra F - Clin. Sci. (2012)

Bottom Line: Sirius Red staining demonstrated pericellular fibrosis in zone 3, and image analysis showed a significantly lower matrix accumulation in CCL2-KO mice.This was associated with reduced hepatic expression of TGF-β (transforming growth factor-β), type I procollagen, TIMP-1 (tissue inhibitor of metalloproteinases-1) and α-smooth muscle actin.We conclude that, in experimental murine steatohepatitis, the effects of CCL2 deficiency are markedly dependent on the genetic background.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Medicina Interna, University of Florence, Florence, Italy.

ABSTRACT
Expression of CCL2 (CC chemokine ligand 2) (or monocyte chemoattractant protein-1) regulates inflammatory cell infiltration in the liver and adipose tissue, favouring steatosis. However, its role in the pathogenesis of steatohepatitis is still uncertain. In the present study, we investigated the development of non-alcoholic steatohepatitis induced by an MCD diet (methionine/choline-deficient diet) in mice lacking the CCL2 gene on two different genetic backgrounds, namely Balb/C and C57/Bl6J. WT (wild-type) and CCL2-KO (knockout) mice were fed on a lipid-enriched MCD diet or a control diet for 8 weeks. In Balb/C mice fed on the MCD diet, a lack of CCL2 was associated with lower ALT (alanine transaminase) levels and reduced infiltration of inflammatory cells, together with a lower generation of oxidative-stress-related products. Sirius Red staining demonstrated pericellular fibrosis in zone 3, and image analysis showed a significantly lower matrix accumulation in CCL2-KO mice. This was associated with reduced hepatic expression of TGF-β (transforming growth factor-β), type I procollagen, TIMP-1 (tissue inhibitor of metalloproteinases-1) and α-smooth muscle actin. In contrast, in mice on a C57Bl/6 background, neither ALT levels nor inflammation or fibrosis were significantly different comparing WT and CCL2-KO animals fed on an MCD diet. In agreement, genes related to fibrogenesis were expressed to comparable levels in the two groups of animals. Comparison of the expression of several genes involved in inflammation and repair demonstrated that IL (interleukin)-4 and the M2 marker MGL-1 (macrophage galactose-type C-type lectin 1) were differentially expressed in Balb/C and C57Bl/6 mice. No significant differences in the degree of steatosis were observed in all groups of mice fed on the MCD diet. We conclude that, in experimental murine steatohepatitis, the effects of CCL2 deficiency are markedly dependent on the genetic background.

Show MeSH

Related in: MedlinePlus

Lack of CCL2 reduces aminotransferase levels in Balb/C miceWT or CCL2-KO mice were fed for 8 weeks on the control diet or on the MCD diet. WT Cnt, WT animals fed on the control diet; KO Cnt, CCL2-KO animals fed on the control diet; WT MCD, WT animals fed on the MCD diet; KO MCD, CCL2-KO animals fed on the MCD diet. At the end of the study protocol, animals were killed and serum ALT (A) and AST (B) were assayed as described in Materials and methods section. *P<0.01 compared with WT animals fed on the control diet; **P<0.01 compared with WT animals fed on the MCD diet. UI, international units.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3369401&req=5

Figure 1: Lack of CCL2 reduces aminotransferase levels in Balb/C miceWT or CCL2-KO mice were fed for 8 weeks on the control diet or on the MCD diet. WT Cnt, WT animals fed on the control diet; KO Cnt, CCL2-KO animals fed on the control diet; WT MCD, WT animals fed on the MCD diet; KO MCD, CCL2-KO animals fed on the MCD diet. At the end of the study protocol, animals were killed and serum ALT (A) and AST (B) were assayed as described in Materials and methods section. *P<0.01 compared with WT animals fed on the control diet; **P<0.01 compared with WT animals fed on the MCD diet. UI, international units.

Mentions: We first analysed the effects of an MCD diet given for 8 weeks to WT and CCL2-KO Balb/C mice. At the end of the treatment period, mice fed on the MCD diet showed a marked and significant decrease in body weight (Table 2). However, no differences were observed comparing WT and CCL2-KO mice with either diet regimen. The liver/body weight ratio tended to be higher in mice fed on the MCD diet (Table 2), but no significant differences were found comparing the different diets or the different mouse genotypes. In WT mice fed on the MCD diet, the levels of ALT and AST (aspartate aminotransferase) were increased more than 5-fold in comparison with mice treated with the control diet (Figure 1). In contrast, in mice lacking CCL2, aminotransferase levels were significantly lower than in WT animals fed on the MCD diet. No differences were observed comparing WT and CCL2-KO mice fed on the control diet.


Lack of CC chemokine ligand 2 differentially affects inflammation and fibrosis according to the genetic background in a murine model of steatohepatitis.

Galastri S, Zamara E, Milani S, Novo E, Provenzano A, Delogu W, Vizzutti F, Sutti S, Locatelli I, Navari N, Vivoli E, Caligiuri A, Pinzani M, Albano E, Parola M, Marra F - Clin. Sci. (2012)

Lack of CCL2 reduces aminotransferase levels in Balb/C miceWT or CCL2-KO mice were fed for 8 weeks on the control diet or on the MCD diet. WT Cnt, WT animals fed on the control diet; KO Cnt, CCL2-KO animals fed on the control diet; WT MCD, WT animals fed on the MCD diet; KO MCD, CCL2-KO animals fed on the MCD diet. At the end of the study protocol, animals were killed and serum ALT (A) and AST (B) were assayed as described in Materials and methods section. *P<0.01 compared with WT animals fed on the control diet; **P<0.01 compared with WT animals fed on the MCD diet. UI, international units.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3369401&req=5

Figure 1: Lack of CCL2 reduces aminotransferase levels in Balb/C miceWT or CCL2-KO mice were fed for 8 weeks on the control diet or on the MCD diet. WT Cnt, WT animals fed on the control diet; KO Cnt, CCL2-KO animals fed on the control diet; WT MCD, WT animals fed on the MCD diet; KO MCD, CCL2-KO animals fed on the MCD diet. At the end of the study protocol, animals were killed and serum ALT (A) and AST (B) were assayed as described in Materials and methods section. *P<0.01 compared with WT animals fed on the control diet; **P<0.01 compared with WT animals fed on the MCD diet. UI, international units.
Mentions: We first analysed the effects of an MCD diet given for 8 weeks to WT and CCL2-KO Balb/C mice. At the end of the treatment period, mice fed on the MCD diet showed a marked and significant decrease in body weight (Table 2). However, no differences were observed comparing WT and CCL2-KO mice with either diet regimen. The liver/body weight ratio tended to be higher in mice fed on the MCD diet (Table 2), but no significant differences were found comparing the different diets or the different mouse genotypes. In WT mice fed on the MCD diet, the levels of ALT and AST (aspartate aminotransferase) were increased more than 5-fold in comparison with mice treated with the control diet (Figure 1). In contrast, in mice lacking CCL2, aminotransferase levels were significantly lower than in WT animals fed on the MCD diet. No differences were observed comparing WT and CCL2-KO mice fed on the control diet.

Bottom Line: Sirius Red staining demonstrated pericellular fibrosis in zone 3, and image analysis showed a significantly lower matrix accumulation in CCL2-KO mice.This was associated with reduced hepatic expression of TGF-β (transforming growth factor-β), type I procollagen, TIMP-1 (tissue inhibitor of metalloproteinases-1) and α-smooth muscle actin.We conclude that, in experimental murine steatohepatitis, the effects of CCL2 deficiency are markedly dependent on the genetic background.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Medicina Interna, University of Florence, Florence, Italy.

ABSTRACT
Expression of CCL2 (CC chemokine ligand 2) (or monocyte chemoattractant protein-1) regulates inflammatory cell infiltration in the liver and adipose tissue, favouring steatosis. However, its role in the pathogenesis of steatohepatitis is still uncertain. In the present study, we investigated the development of non-alcoholic steatohepatitis induced by an MCD diet (methionine/choline-deficient diet) in mice lacking the CCL2 gene on two different genetic backgrounds, namely Balb/C and C57/Bl6J. WT (wild-type) and CCL2-KO (knockout) mice were fed on a lipid-enriched MCD diet or a control diet for 8 weeks. In Balb/C mice fed on the MCD diet, a lack of CCL2 was associated with lower ALT (alanine transaminase) levels and reduced infiltration of inflammatory cells, together with a lower generation of oxidative-stress-related products. Sirius Red staining demonstrated pericellular fibrosis in zone 3, and image analysis showed a significantly lower matrix accumulation in CCL2-KO mice. This was associated with reduced hepatic expression of TGF-β (transforming growth factor-β), type I procollagen, TIMP-1 (tissue inhibitor of metalloproteinases-1) and α-smooth muscle actin. In contrast, in mice on a C57Bl/6 background, neither ALT levels nor inflammation or fibrosis were significantly different comparing WT and CCL2-KO animals fed on an MCD diet. In agreement, genes related to fibrogenesis were expressed to comparable levels in the two groups of animals. Comparison of the expression of several genes involved in inflammation and repair demonstrated that IL (interleukin)-4 and the M2 marker MGL-1 (macrophage galactose-type C-type lectin 1) were differentially expressed in Balb/C and C57Bl/6 mice. No significant differences in the degree of steatosis were observed in all groups of mice fed on the MCD diet. We conclude that, in experimental murine steatohepatitis, the effects of CCL2 deficiency are markedly dependent on the genetic background.

Show MeSH
Related in: MedlinePlus