The effect of ADAM8 expression on acute alcoholic liver injury in mice

doi:10.11958/20231741

Abstract

Abstract:

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.

Key words: ADAM proteins, acute alcoholic liver injury, mitogen-activated protein kinases, CRISPR/Cas9

CLC Number:

R364.5

Figures/Tables 5

References 14

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组别	n	肝损伤评分/分	糖原光密度比值
正常组	6	0.00±0.00	1.00±0.60
酒精组	6	5.00±2.00^a	0.00±0.00^a
质粒组	6	1.67±0.58^b	0.04±0.00^a
F		13.460^＊＊	7.911^＊

组别	n	肝损伤评分/分	糖原光密度比值
正常组	6	0.00±0.00	1.00±0.60
酒精组	6	5.00±2.00^a	0.00±0.00^a
质粒组	6	1.67±0.58^b	0.04±0.00^a
F		13.460^＊＊	7.911^＊

组别	n	ALT/（U/L）	AST/（U/L）
正常组	6	13.09±2.05	16.78±1.97
酒精组	6	24.48±5.88^a	38.54±13.36^a
质粒组	6	15.40±1.63^b	18.71±6.14^b
F		15.750^＊＊	11.870^＊＊

组别	n	ALT/（U/L）	AST/（U/L）
正常组	6	13.09±2.05	16.78±1.97
酒精组	6	24.48±5.88^a	38.54±13.36^a
质粒组	6	15.40±1.63^b	18.71±6.14^b
F		15.750^＊＊	11.870^＊＊

组别	ADAM8	CYP2E1	HSP70
正常组	1.00±0.00	1.00±0.00	1.00±0.00
酒精组	1.74±0.32^a	2.30±0.58^a	0.50±0.08^a
质粒组	0.69±0.30^b	0.77±0.55^b	0.71±0.11^b
F	13.540^＊＊	9.690^＊	30.240^＊＊
组别	p-ERK1/2	p-p38	p-c-Jun
正常组	1.00±0.00	1.00±0.00	1.00±0.00
酒精组	1.45±0.12^a	2.06±0.50^a	1.71±0.25^a
质粒组	0.84±0.16^b	1.10±0.42^b	1.19±0.11^b
F	22.170^＊＊	7.350^＊	16.330^＊＊