Targeting VEGF using Bevacizumab attenuates sepsis-induced liver injury in a mouse model of cecal ligation and puncture

Abstract

Sepsis, a life-threatening condition resulting from an uncontrolled host response to infection, often leads to severe liver damage and remains a significant cause of mortality in critically ill patients despite advances in antibiotic therapy and resuscitation. Bevacizumab, a neutralizing antibody targeting vascular endothelial growth factor (VEGF), is approved for treating certain cancers. However, its potential impact on sepsis-related liver injury is not well understood. This study aimed to explore the potential hepatoprotective effect of Bevacizumab on sepsis-induced liver injury. Twenty-four mice were divided into four groups: a sham group subjected to a midline incision only, a cecal ligation and puncture induction (CLP) group, a vehicle-treated group that received a vehicle one hour before CLP induction, and a Bevacizumab-treated group that received Bevacizumab one hour before CLP induction. Blood samples were collected, and angiopoietin-2 (ANGPT2), alanine transaminase (ALT), and aspartate transaminase (AST) serum levels were measured. Liver tissue homogenates were analyzed for IL-6, TNFα, intracellular adhesion molecule (ICAM-1), macrophage inhibitory factor (MIF), vascular endothelial growth factor (VEGF), F2-isoprostane, and caspase-11 levels. A histological examination was performed to assess the extent of liver damage. Mice exposed to CLP had high levels of the biomarkers mentioned above with a high degree of liver injury compared to the sham group. In contrast, treatment with Bevacizumab notably reduced these markers and mitigated liver damage. In conclusion, Bevacizumab may be a protective agent against sepsis-induced liver injury.

Keywords: Bevacizumab; VEGF; mouse model; sepsis-induced liver injury.

Figure 1

Effect of Bevacizumab on AST and ALT levels following CLP-induced liver injury Serum levels of AST and ALT across the four groups were measured in U/L. Data are presented as mean±SEM, n=6. One-way ANOVA and Bonferroni test.***p≤0.001, ****p≤0.0001

Figure 2

Effect of Bevacizumab on TNF-α and IL-6 levels following CLP Levels of TNF-α and IL-6 in the tissues of the liver across the four groups were measured in ng/L. Results are presented as mean±SEM, n=6. One-way ANOVA and Bonferroni test, **p≤0.01.

Figure 3

Effect of Bevacizumab on F2-isoprostane levels following CLP The levels of F2-isoprostane in the liver across the four groups were measured in ng/L. Results are presented as mean ±SEM, n=6. One-way ANOVA and Bonferroni test, ***p≤0.001.

Figure 4

Effect of Bevacizumab on caspase-11 levels following CLP The levels of caspase-11 in the liver across the four groups were measured in ng/L. The results are presented as mean±SEM, n=6. One-way ANOVA and Bonferroni test, **p≤0.01.

Figure 5

Effect of Bevacizumab on MIF levels following CLP The levels of MIF in the tissues of the liver across the four groups were measured in ng/L. The results are expressed as mean±SEM, n=6. One-way ANOVA and Bonferroni test, ****p≤0.0001.

Figure 6

Effect of Bevacizumab on ANGPT2 levels following CLP Serum levels of ANGPT2 across the four groups were measured in ng/L. Data are presented as mean±SEM, n=6. One-way ANOVA and Bonferroni test.***p≤0.001.

Figure 7

Effect of Bevacizumab on ICAM-1 levels following CLP ICAM-1 levels in the tissues of the liver across the four study groups were measured in ng/L. The results are presented as mean ±SEM, n=6. One-way ANOVA and Bonferroni test, ****p≤0.0001.

Figure 8

Effect of Bevacizumab on VEGF levels following CLP VEGF levels in liver tissues across the four study groups were measured in ng/L. The results are presented as mean±SEM, n=6. One-way ANOVA and Bonferroni test, ****p≤0.0001.

Figure 9

Histological liver damage scores Mean histological scores representing liver damage across four experimental groups. Data presented as mean±SEM, n=6. Kruskal-Wallis test ***p≤0.001.

Figure 10

Liver histology in sham group. A: Normal liver histology in the sham group (X 100 magnification). B: Central vein (red arrow) and normal hepatocytes (black arrow) in the sham group (X 400 magnification).

Figure 11

Liver sections showing the morphological changes in the CLP group. A: Severe liver damage in liver sections (score 3) (X100 magnification). B: Cytoplasmic eosinophilia, apoptotic cells with chromatin condensation, pyknotic nuclei (black arrows) (X400 magnification). C: Necrotic cells with nuclear fading (black arrows) (X400 magnification). D: Vascular congestion with erythrocyte stasis (black arrows) (X400 magnification). E: Cytoplasmic vacuoles (X400 magnification).

Figure 12

Liver sections showing the impact of Bevacizumab treatment on liver tissues following CLP. A: Mild liver damage in liver sections (score 1) (X100 magnification). B: Vascular congestion with erythrocyte stasis (red arrows) (X400 magnification). C: Vascular congestion with erythrocyte stasis (black arrows), cytoplasmic vacuoles, and steatosis (red arrows) (X400 magnification).