Mechanism of inhibiting a disintegrin and metalloprotease 8 expression on inflammatory damage in alcoholic liver fibrosis mice
10.16098/j.issn.0529-1356.2024.06.013
- VernacularTitle:抑制解整合素金属蛋白酶8表达对酒精性肝纤维化小鼠炎症损伤的机制
- Author:
Meng-Li YANG
1
;
San-Qiang LI
;
Kai-Jie ZHANG
;
Jia-Yang FENG
;
Hao-Yuan LI
;
Chang XU
Author Information
1. 河南科技大学基础医学与法医学院肝脏损伤与修复分子医学重点实验室,河南洛阳 471000;河南省肝病防治工程技术研究中心,河南洛阳 471000
- Keywords:
A disintegrin and metalloprotease 8;
Alcoholic liver fibrosis;
Inflammation;
Damage;
Transforming growth factor-β1/p38 mitogenactivated protein kinase signaling pathway;
Western blotting;
Mouse
- From:
Acta Anatomica Sinica
2024;55(6):746-752
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the mechanism of inhibiting a disintegrin and metalloprotease 8(ADAM8)expression on inflammatory damage in alcoholic liver fibrosis mice.Methods C57BL/6N male mice were randomly divided into the control group,the alcohol group,and the plasmid group,with 10 mice in each group.The alcohol group and plasmid group were fed alcohol liquid feed on a daily basis and gavaged with 31.5％ethanol(5 g/kg,twice a week);The control group was fed control liquid feed and gavaged with an equal amount of saline.The plasmid group was injected with the effective plasmid ADAM8-small guide RNA3(sgRNA3)(2 g/kg,twice a week)to inhibit the ADAM8 gene through the tail vein,while the alcohol group was injected with an equal amount of saline through the tail vein for 8 weeks to induce alcoholic liver fibrosis.After eyeball blood collection,the mice were euthanized,and their liver was separated and extracted.Sirius red and HE staining were employed to assess liver fibrosis and damage;Western blotting was used to determine the expression of α-smooth muscle actin(α-SMA),collagen Ⅰ,and ADAM8;Real-time PCR was used to measure the expression of ADAM8 mRNA;the expression of ADAM8,transforming growth factor-β1(TGF-β1),p-p38 MAPK and heat shock protein 27(HSP27)proteins was detected by Western blotting,Biochemical detection were used to detect alanine aminotransferase(ALT)and aspartate transaminase(AST)activity;tumor necrosis factor-α(TNF-α)and interleukin-1(IL-1)levels were determined by ELISA.Results The alcohol group had increased collagen fiber volume fraction,liver injury scores,and positive area rates of α-SMA and collagen Ⅰ;the expression of ADAM8 mRNA and ADAM8 protein increased,with increased positive area rate;and levels of AST,ALT,TNF-α,and IL-1 were higher,along with increased expression of TGF-β1,p-p38MAPK,and HSP27 proteins.In the plasmid group,the collagen fiber volume fraction,liver injury scores,and positive area rates of α-SMA and collagen Ⅰ was reduced;the expression of ADAM8 mRNA and protein was reduced,with decreased positive area rate,and levels of AST,ALT,TNF-α,and IL-1 were lower,along with reduced expression of TGF-β1,p-p38MAPK,and HSP27 proteins.Conclusion Downregulation of ADAM8 expression can alleviate inflammatory damage by inhibiting TGF-β1/p38MAPK signaling pathway to improve alcoholic liver fibrosis in mice.
