The effect of ADAM8 expression on acute alcoholic liver injury in mice
Objective To investigate the role and molecular mechanism of CRISPR/Cas9 technology targeting the inhibition of disintegrin and metalloprotease 8(ADAM8)in acute alcoholic liver injury in mice.Methods Eighteen healthy Kunming female mice aged 6-8 weeks were randomly divided into the normal group,the alcohol group and the plasmid group,with 6 mice in each group.The normal group was given no treatment,and the plasmid group was injected with three-in-one recombinant plasmid ADAM8-sgRNA3(3 g/kg)by CRISPR/Cas9 technology to inhibit the ADAM8 gene using hydrodynamic injection tail vein method.The alcohol group was injected with an equal amount of physiological saline through tail vein.After 3 days of regular feeding,the alcohol group and the plasmid group were subjected with 50%(V/V)alcohol analytical grade ethanol by a one-time gavage(14 mL/kg)to induce acute liver injury.After fasting for 16 hours,eyeball blood sample was taken to detect activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST).Mice were euthanized and liver tissue was separated and extracted.Hematoxylin-Eosin staining(HE staining)and Periodic Acid-Schiff staining(PAS staining)were used to detect liver injury and glycogen changes in each group of mice.The expression levels of ADAM8,cytochrome CYP450 2E1(CYP2E1),heat shock protein 70(HSP70)and mitogen-activated protein kinase(MAPK)pathway related factors were detected by Western blot assay.Results Compared with the normal group,ALT and AST were increased,liver injury score was increased,glycogen content was decreased,ADAM8,CYP2E1,phosphorylated extracellular regulated kinase 1/2(p-ERK1/2),phosphorylated p38(p-p38)MAPK and phosphorylated c-Jun aminoterminal kinase(p-c-Jun)were increased in the alcohol group.The expression of HSP70 was decreased(P＜0.05).Compared with alcohol group,ALT and AST were decreased,liver injury score was decreased in the plasmid group(P＜0.05).Glycogen content was increased.ADAM8,CYP2E1,p-ERK1/2,p-P38 MAPK and p-c-Jun expression levels were decreased,while HSP70 expression was increased(P＜0.05).Conclusion Targeted inhibition of ADAM8 expression by CRISPR/Cas9 technology can improve acute alcoholic liver injury in mice through MAPK signaling pathway.
ADAM proteinsacute alcoholic liver injurymitogen-activated protein kinasesCRISPR/Cas9
万方数据
维普
