Effects of Adeno-associated Virus Mediated Overexpression of Trx1 on Cadmium Exposure Induced Liver Injury and Ferropto-sis in Mice

CAO Zhengwang; LIU Kun; XIAO Hongyu; ZHENG Shan'gen; YU Tiantian

doi:10.13730/j.issn.2097-2148.2025.02.001

Abstract：Objective To investigate the effects of adeno-associated virus(AAV)upregulation of thioredoxin1(Trx1)expression on Cadmium(Cd)exposure induced liver injury and hepatocyte ferroptosis in mice.Methods A total of 24 male C57BL/6J mice were randomly divided into control group,Trx1 group,Cd group and Trx1+Cd group,with 6 mice in each.Animal model of Trx1 overexpression was established via tail vein injection of AAV,and Cd exposure model was created by intraperitoneal injection of 2 mg/kg cadmium chloride(CdCl2).After 24 h of Cd ex-posure,the mice were anesthetized and excuted.Trx1 ex-pression in the mice liver was underwent immunofluores-cence staining and then was detected by Western blot and quantitative real-time polymerase chain reaction(qPCR).The concentration of alanine aminotransferase(ALT),aspar-tate aminotransferase(AST),lactate dehydrogenase(LDH)and total bilirubin(TBIL)in serum were measured;the con-centration of malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)in liver tissues were detected;hepatic pathological changes were observed by hematoxylin-eosin staining,mitochon-drial morphology in hepatocytes was examined by transmission electron microscopy;the expression of ferroptosis-related proteins,including glutathione peroxidase 4(GPX4),nuclear factor erythroid-2-related factor 2(NRF2),heme oxygen-ase 1(HO-1),solute carrier family 7 member 11(SLC7A11)and prostaglandin endoperoxide synthase 2(PTGS2)in liv-er tissues of the mice were detected by Western blot.Results Compared with the control group,the expression levels of Trx1 protein and mRNA in liver tissues of Trx1 group significantly increased(P＜0.05).Compared with the mice in Cd group,the levels of ALT,AST and LDH in serum and MDA level in liver tissues of Trx1+Cd group decreased,while the contents of SOD,CAT and GSH-Px increased(all P＜0.05).The degree of hepatocyte necrosis and inflammatory in-filtration was reduced,the degree of mitochondrial shrinkage,bilayer membrane density and mitochondrial matrix swell-ing were alleviated.Compared with Cd group,the protein expression levels of GPX4,NRF2,HO-1 and SLC7A11 in liver tissues of Trx1+Cd group increased(all P＜0.05),while PTGS2 protein expression level decreased(P＜0.01).Conclusion AAV overexpressed Trx1 can alleviate Cd exposure induced liver injury and oxidative stress,the mechanism may be related to the suppression of ferroptosis by the SLC7A11/GPX4 signaling pathway.