Effects Of KLHL15 Gene On Liver And Kidney Damage In Mice Exposed To Lead

Background: More than 40 kinds of Kelch proteins have been found and have been proved to play a role in regulating protein interaction,protein degradation,gene expression and signal transduction,and biological morphology regulation as substrates of E3 ubiquitin ligase.In 2004,Kenichi Yoshida identified the human homologous KLHL15 gene of Drosophila melanogaster for the first time by using bioinformatics methods,determined the complete sequence of human KLHL15 c DNA,and revealed that the m RNA of human KLHL15 was ubiquitously expressed in various tissues.At present,it has been confirmed that KLHL15 and Cullin3 proteins constitute E3 ubiquitin ligase,which affects the process of protein ubiquitination and continues to act on downstream substrates,such as B ’that mediates protein Ser/Thr phosphatase 2A(PP2A)β.It regulates the ubiquitination of subunits and subsequent proteasome degradation,which may affect cell division,cell homeostasis and apoptosis.KLHL15 can promote the ubiquitination and degradation of DNA repair protein Ct IP to regulate DNA terminal excision,which can affect DNA repair ability.In this study,C57BL/6J mice with KLHL15 gene knockout were used to explore the damage of liver and kidney tissue of mice after KLHL15 knockout under the condition of lead acetate exposure,so as to provide valuable information for further understanding the role of KLHL15 in tissue damage and cell apoptosis.Objective: In this study,the KLHL15 gene knockout C57BL/6J mice were used to explore the damage and potential biological mechanism of lead acetate on the liver and kidney tissues of the gene knockout mice,so as to provide valuable information for further understanding the biological function of KLHL15 in tissue damage and cell apoptosis.Methods: 1.Reproduction,mouse tail genotyping and screening of KLHL15 gene knockout homozygote mice.2.The liver and kidney injury models of mice induced by lead were established by intragastric administration,including wild type(WT)mice,wild type lead treated mice,KLHL15 gene knockout(KO)mice and KLHL15 gene knockout lead treated mice.3.The body weight and organ coefficient of mice in each group were recorded and compared,and the growth of mice was observed.4.The levels of total superoxide dismutase(T-SOD),glutathione peroxidase(GSH-PX),catalase activity(CAT)and malondialdehyde(MDA)in the liver and kidney of mice in each group were detected to compare the antioxidant capacity of liver and kidney tissues of mice in each group;HE staining was used to detect the pathological changes in the liver and kidney tissues of mice,and to compare the tissue damage of each group of mice;TUNEL method was used to detect the apoptosis of mouse liver and kidney cells.5.Qualitative,quantitative and bioinformatics analysis of mouse liver protein in each group was performed by TMT-labeled quantitative proteome analysis.Results : 1.This study found that knocking out the KLHL15 gene significantly reduced the liver and kidney index of lead exposed mice.2.By measuring the levels of T-SOD,GSH-PX,CAT and MDA in the liver and kidney,this study found that lead acetate can cause a decrease in the antioxidant capacity of liver and kidney tissues,and knocking out the KLHL15 gene can further reduce the antioxidant capacity of liver and kidney tissues.3.By observing the results of HE staining and TUNEL cell apoptosis,this study revealed that with the increase of lead acetate dosage,the damage and apoptosis of mouse liver and kidney histiocyte showed an increasing trend,and KLHL15 gene knockout further aggravated the damage and apoptosis of liver and kidney tissue.4.Quantitative proteomic analysis based on TMT showed that in lead-induced mice,KLHL15 gene knockout mainly affected the cell structure including mitochondria,peroxisome and peroxisome membrane,and the activity of partial oxidoreductase,and played a key role in some signal pathways,such as glycine,serine and threonine metabolism,drug metabolism,peroxisome proliferator-activated receptor(PPAR)signal pathway,etc.Conclusion: KLHL15 gene knockout aggravated lead induced liver and kidney damage in mice,which may be caused by apoptosis and oxidative stress of liver and kidney cells through peroxisome proliferator activated receptor(PPAR)signaling pathway and glycine,serine and threonine metabolic pathway.