Liver fibrosis is a wound-healing response against chronic insult that leads to an alteration of the homeostasis within the hepatic parenchyma. The main contributors of this disequilibrium are the inflammatory processes which activate Hepatic Stellate Cells (HSCs), sinusoidal cells known as the principal triggers of fibrogenesis. Advanced stages of liver fibrosis result in cirrhosis, an irreversible condition of liver damage, and finally hepatocellular carcinoma. The aim of this work is to investigate gender-related differences between male and female mice during liver fibrosis and regeneration, focusing on the regulation of the main immune cells, resident in or infiltrating the liver. To induce liver fibrosis, increasing doses of carbon tetrachloride (CCl4, from 0.17 to 0.72 uL/g body weight) have been intraperitoneally injected to Balb/C mice for 12 weeks, followed by an 8-week washout period to allow liver regeneration. Mice were then sacrificed at week 6, 12 and 20. In order to identify the entity of damage, liver histology was assessed by Masson’s trichrome staining. Liver inflammation was evaluated by measuring the gene expression of pro-inflammatory cytokines, while the collagen content and the activation of HSCs, measured by the expression of their marker alpha-smooth muscle actin (-SMA) were evaluated by Western Blot and Immunohistochemistry, respectively. Flow cytometry experiments were performed on fresh liver tissues to calculate the amount of recruited monocytes-derived macrophages (MoMFs), neutrophils, and Kupffer cells. The results obtained highlighted that this new model of increasing doses of CCl4 led to the development of an increasing liver damage without adaptive responses of the liver to the injury. In addition, it has been observed that gender influences the recruitment of pro-inflammatory immune cells and the activation of hepatic stellate cells. Specifically, the damage occurs more rapidly in males than females, while females recover better than males from liver damage. Furthermore, we noticed a different recruitment of immune cells between males and females. In particular, neutrophil levels are higher in males than females in the whole period of treatment and washout, while Kupffer cell levels are initially equal between genders, and then they become higher in males.

Liver fibrosis is a wound-healing response against chronic insult that leads to an alteration of the homeostasis within the hepatic parenchyma. The main contributors of this disequilibrium are the inflammatory processes which activate Hepatic Stellate Cells (HSCs), sinusoidal cells known as the principal triggers of fibrogenesis. Advanced stages of liver fibrosis result in cirrhosis, an irreversible condition of liver damage, and finally hepatocellular carcinoma. The aim of this work is to investigate gender-related differences between male and female mice during liver fibrosis and regeneration, focusing on the regulation of the main immune cells, resident in or infiltrating the liver. To induce liver fibrosis, increasing doses of carbon tetrachloride (CCl4, from 0.17 to 0.72 uL/g body weight) have been intraperitoneally injected to Balb/C mice for 12 weeks, followed by an 8-week washout period to allow liver regeneration. Mice were then sacrificed at week 6, 12 and 20. In order to identify the entity of damage, liver histology was assessed by Masson’s trichrome staining. Liver inflammation was evaluated by measuring the gene expression of pro-inflammatory cytokines, while the collagen content and the activation of HSCs, measured by the expression of their marker alpha-smooth muscle actin (-SMA) were evaluated by Western Blot and Immunohistochemistry, respectively. Flow cytometry experiments were performed on fresh liver tissues to calculate the amount of recruited monocytes-derived macrophages (MoMFs), neutrophils, and Kupffer cells. The results obtained highlighted that this new model of increasing doses of CCl4 led to the development of an increasing liver damage without adaptive responses of the liver to the injury. In addition, it has been observed that gender influences the recruitment of pro-inflammatory immune cells and the activation of hepatic stellate cells. Specifically, the damage occurs more rapidly in males than females, while females recover better than males from liver damage. Furthermore, we noticed a different recruitment of immune cells between males and females. In particular, neutrophil levels are higher in males than females in the whole period of treatment and washout, while Kupffer cell levels are initially equal between genders, and then they become higher in males.

Immunological orchestration of liver fibrosis and regeneration in male and female mice

DANIELI, GIULIA
2021/2022

Abstract

Liver fibrosis is a wound-healing response against chronic insult that leads to an alteration of the homeostasis within the hepatic parenchyma. The main contributors of this disequilibrium are the inflammatory processes which activate Hepatic Stellate Cells (HSCs), sinusoidal cells known as the principal triggers of fibrogenesis. Advanced stages of liver fibrosis result in cirrhosis, an irreversible condition of liver damage, and finally hepatocellular carcinoma. The aim of this work is to investigate gender-related differences between male and female mice during liver fibrosis and regeneration, focusing on the regulation of the main immune cells, resident in or infiltrating the liver. To induce liver fibrosis, increasing doses of carbon tetrachloride (CCl4, from 0.17 to 0.72 uL/g body weight) have been intraperitoneally injected to Balb/C mice for 12 weeks, followed by an 8-week washout period to allow liver regeneration. Mice were then sacrificed at week 6, 12 and 20. In order to identify the entity of damage, liver histology was assessed by Masson’s trichrome staining. Liver inflammation was evaluated by measuring the gene expression of pro-inflammatory cytokines, while the collagen content and the activation of HSCs, measured by the expression of their marker alpha-smooth muscle actin (-SMA) were evaluated by Western Blot and Immunohistochemistry, respectively. Flow cytometry experiments were performed on fresh liver tissues to calculate the amount of recruited monocytes-derived macrophages (MoMFs), neutrophils, and Kupffer cells. The results obtained highlighted that this new model of increasing doses of CCl4 led to the development of an increasing liver damage without adaptive responses of the liver to the injury. In addition, it has been observed that gender influences the recruitment of pro-inflammatory immune cells and the activation of hepatic stellate cells. Specifically, the damage occurs more rapidly in males than females, while females recover better than males from liver damage. Furthermore, we noticed a different recruitment of immune cells between males and females. In particular, neutrophil levels are higher in males than females in the whole period of treatment and washout, while Kupffer cell levels are initially equal between genders, and then they become higher in males.
2021
Immunological orchestration of liver fibrosis and regeneration in male and female mice
Liver fibrosis is a wound-healing response against chronic insult that leads to an alteration of the homeostasis within the hepatic parenchyma. The main contributors of this disequilibrium are the inflammatory processes which activate Hepatic Stellate Cells (HSCs), sinusoidal cells known as the principal triggers of fibrogenesis. Advanced stages of liver fibrosis result in cirrhosis, an irreversible condition of liver damage, and finally hepatocellular carcinoma. The aim of this work is to investigate gender-related differences between male and female mice during liver fibrosis and regeneration, focusing on the regulation of the main immune cells, resident in or infiltrating the liver. To induce liver fibrosis, increasing doses of carbon tetrachloride (CCl4, from 0.17 to 0.72 uL/g body weight) have been intraperitoneally injected to Balb/C mice for 12 weeks, followed by an 8-week washout period to allow liver regeneration. Mice were then sacrificed at week 6, 12 and 20. In order to identify the entity of damage, liver histology was assessed by Masson’s trichrome staining. Liver inflammation was evaluated by measuring the gene expression of pro-inflammatory cytokines, while the collagen content and the activation of HSCs, measured by the expression of their marker alpha-smooth muscle actin (-SMA) were evaluated by Western Blot and Immunohistochemistry, respectively. Flow cytometry experiments were performed on fresh liver tissues to calculate the amount of recruited monocytes-derived macrophages (MoMFs), neutrophils, and Kupffer cells. The results obtained highlighted that this new model of increasing doses of CCl4 led to the development of an increasing liver damage without adaptive responses of the liver to the injury. In addition, it has been observed that gender influences the recruitment of pro-inflammatory immune cells and the activation of hepatic stellate cells. Specifically, the damage occurs more rapidly in males than females, while females recover better than males from liver damage. Furthermore, we noticed a different recruitment of immune cells between males and females. In particular, neutrophil levels are higher in males than females in the whole period of treatment and washout, while Kupffer cell levels are initially equal between genders, and then they become higher in males.
Liver
Fibrosis
Immune cells
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12608/35920