Protective effect of overexpression of miR-140-5p mediated by lentivirus on lipopolysaccharide-induced liver injury in rats
10.3969/j.issn.1000-484X.2025.01.011
- VernacularTitle:慢病毒介导miR-140-5p过表达对内毒素诱导大鼠肝损伤的保护作用
- Author:
Li LI
1
;
Haishan CUI
1
;
Mei LI
1
Author Information
1. 内蒙古民族大学附属医院消化二科,通辽 028000
- Publication Type:Journal Article
- Keywords:
miR-140-5p;
Endotoxin;
Liver injury;
Inflammatory reaction;
Oxidative stress
- From:
Chinese Journal of Immunology
2025;41(1):69-74
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the protective effect of lentivirus-mediated miR-140-5p overexpression on lipopolysaccha-ride-induced liver injury in rats.Methods:A total of 40 rats were randomly divided into control group,model group,miR-NC group and miR-140-5p-mimic group,with 10 rats in each group.Rats in miR-NC group and miR-140-5p-mimic group received tail vein injection of lentivirus empty vector miR-NC and recombinant miR-140-5p respectively,while rats in control group and model group received tail vein injection of normal saline.Three days later,rats in model group,miR-NC group and miR-140-5p-mimic group received intraperi-toneal injection of 10 mg/kg lipopolysaccharide,while rats in control group received intraperitoneal injection of normal saline.All rats were killed 12 hours after injection,and the serum levels of ALT,AST,endotoxin,TNF-α,IL-6,IL-1β,SOD,MDA and CAT were detected.Pathological morphology and apoptosis of liver tissue were observed by HE and TUNEL staining.TargetScanHuman was used to predicted binding site of miR-140-5p and TLR4,and double luciferase reporter gene assay was used to analyse the targeted regula-tion relationship between miR-140-5p and TLR4.Level of miR-140-5p in liver tissue was detected by qRT-PCR,mRNA and protein expressions of TLR4,MyD88 and NF-κB in liver tissue were detected by qRT-PCR and Western blot,and expression of p-NF-κB was detected by Western blot.Results:In control group,the hepatic lobule was clearly visible,and there was no hepatocyte degeneration,necrosis and inflammatory cell infiltration.Obvious hepatocyte necrosis,vacuolar degeneration,hyperemia and infiltration of inflam-matory cells into hepatic sinusoids were observed in model group and miR-NC group.Hepatocyte hyperemia,necrosis,vacuolar degen-eration and inflammatory cell infiltration in miR-140-5p-mimic group were significantly less than those in model group and miR-NC group.miR-140-5p can directly target TLR4.Compared with control group,levels of ALT,AST,endotoxin,apoptotic cells,TNF-α,IL-6,IL-1β,MDA,TLR4,MyD88 mRNA and protein,NF-κB mRNA,p-NF-κB in model group,miR-NC group and miR-140-5p-mimic group were increased(P<0.05),while SOD and CAT were decreased(P<0.05).Compared with model group and miR-NC group,levels of ALT,AST,endotoxin,apoptotic cells,TNF-α,IL-6,IL-1β,MDA,TLR4,MyD88 mRNA and protein,NF-κB mRNA and p-NF-κB in miR-140-5p-mimic group were decreased(P<0.05),while levels of SOD and CAT were increased(P<0.05).Conclusion:Overexpression of miR-140-5p can reduce the level of inflammatory reaction and oxidative stress in rats with lipopolysac-charide-induced liver injury,protect hepatocytes and reduce liver pathological injury,and its mechanism may be related to TLR4/MyD88/NF-κB signal pathway.
