The Preventive Effect And Mechanism Of Cannabidiol On Acute Liver Injury Induced By CCl₄ In Mice

Objective:To investigate the preventive effect and mechanism of cannabidiol（CBD）on acute liver injury induced by carbon tetrachloride（CCl₄）in mice.Methods:45 6-8 weeks C57BL/6J male mice were divided into 5 groups according to the random number table method:control group,model group,CBD control group,CBD intervention group,and positive control reduced glutathione（GSH）intervention group,the CBD control group,CBD intervention group,and GSH intervention group were intraperitoneally injected with CBD 5mg/kg,CBD 5mg/kg,and GSH 200mg/kg respectively.The control group and the model group were injected with the same dose of normal saline;two hours later,the model group,CBD intervention group and GSH intervention group were intraperitoneally injected with 20%CCl₄olive oil 5m L/kg to establish the acute liver injury model,24 hours later,collect blood after anesthesia,cervical dislocation was executed,and the liver was collected.Detect the levels of serum alanine aminotransferase（ALT）and aspartate aminotransferase（AST）,Liver tissue pathological changes were evaluated by hematoxylin-eosin（HE）staining,Activities of superoxide dismutase（SOD）,malondialdehyde（MDA）,and GSH of liver homogenate were determined.Real-time PCR and western blot detection of liver tissue inflammatory factors:Interleukin-1 beta（IL-1β）,interleukin-6（IL-6）,tumor necrosis factor-α（TNF-α）m RNA and protein expression levels.Western blot was used to detect peroxidase proliferator-activated receptor-γ（PPAR-γ）and nuclear transcription factor kappa-B（NF-κB）,p-NF-κB,mitogen-activated protein kinase p38（MAPK p38）,p-MAPK p38,cyclooxygenase-2（COX-2）protein expression levels.Immunohistochemistry（IHC）was used to detect the expression level of nuclear factor erythroid 2-related factor 2（Nrf2）in liver cells.Results:Compared with the control group,the serum ALT and AST levels of the model group were significantly increased（P<0.05）,HE staining showed that liver cells were damaged and partial necrosis,the levels of SOD and GSH in liver tissue homogenate were significantly reduced（P<0.05）),MDA level was significantly increased（P<0.05）,IL-1β,IL-6,TNF-αm RNA and protein levels were significantly increased（P<0.05）,the protein expression levels of p-NF-κB/NF-κB,p-MAPK p38/MAPK p38,COX-2 were increased（P<0.05）,the expression level of PPAR-γprotein was decreased（P<0.05）,and IHC staining showed that the expression of Nrf2 in hepatocytes was significantly reduced.Compared with the model group,the serum ALT and AST levels of the CBD intervention group and GSH intervention group were significantly reduced（P<0.05）,HE staining showed that the degree and scope of liver cell damage were significantly reduced,and the levels of SOD and GSH in liver tissue homogenate were significantly increased（P<0.05）,MDA level was significantly reduced（P<0.05）,IL-1β,IL-6,and TNF-αm RNA and protein levels were significantly reduced（P<0.05）,p-NF-κB/NF-κB,p-MAPK p38/MAPK p38,COX-2 protein expression levels were reduced（P<0.05）,the expression level of PPAR-γprotein was significantly increased（P<0.05）,IHC staining shows that the expression of Nrf2 in hepatocytes has increased,the CBD control group had no significant difference compared with the control group（P>0.05）.Conclusions:CBD can alleviate the acute liver injury induced by CCl₄in mice,inhibit liver tissue peroxidation and inflammation,its mechanism may be related to up-regulating the expression of PPAR-γand Nrf2.