Forkhead box K1 deficiency alleviates ischemia-reperfusion-induced acute kidney injury
10.3760/cma.j.cn441217-20230818-00827
- VernacularTitle:叉头盒K1缺失减轻缺血再灌注诱导的急性肾损伤
- Author:
Chen LI
1
;
Lu ZHANG
;
Huiming WANG
Author Information
1. 武汉大学人民医院肾内科，武汉 430060
- Keywords:
Reperfusion injury;
Acute kidney injury;
Autophagy;
Forkhead box K1
- From:
Chinese Journal of Nephrology
2024;40(5):379-388
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and mechanism of forkhead box K1 (FOXK1) in acute kidney injury (AKI), and to provide new ideas and targets for preventing and treating AKI.Methods:Three models of AKI were established: 30 male specific pathogen free wild type C57BL/6 mice aged 8-10 weeks and weighting 22-24 g were randomly divided into saline group (0.9% normal saline 0.1 ml/10 g, intraperitoneal injection), lipopolysaccharide (LPS) group (LPS solution 10 mg/kg, intraperitoneal injection), cisplatin group (cisplatin solution 20 mg/kg, intraperitoneal injection), ischemia-reperfusion (IR) group, and sham-operated group by the random number table, with 6 mice in each group. The mice in each group were sacrificed 24 hours after modeling to obtain experimental materials. The serum creatinine (Scr) and blood urea nitrogen (BUN) were tested to measure the renal function. HE staining was performed to observe histopathological changes of renal tissues. Western blotting was used to detect the protein expression of FOXK1 , kidney injury molecule 1 (KIM-1), autophagy markers p62, Beclin1 and LC3 in renal tissues. Quantitative real-time PCR was used to detect the mRNA expression of Foxk1. Human renal tubular epithelial cells (HK-2 cells) were exposed to hypoxia for 24 h, followed by reoxygenation for 6 h to establish an in vitro AKI model induced by hypoxia reoxygenation (HR). The expression changes of the above indicators in HK-2 cells were detected. Then, Foxk1 gene deletion in renal tubular epithelial cells was performed in vivo and in vitro, and AKI models were induced to observe the expression changes of the above indicators. Results:Compared with the saline group, Scr, BUN and the protein expression level of KIM-1 were higher in LPS group and cisplatin group (all P<0.05), while FOXK1 protein and mRNA expression had no significant change (both P>0.05). Compared with the sham-operated group, Scr, BUN and the protein expression level of KIM-1 were higher, and the expression levels of FOXK1 protein and mRNA were significantly lower in the IR group (all P<0.05). FOXK1 protein and mRNA expression levels in the HR-induced AKI cell model group were lower than those in the control group (both P<0.05). In the in vivo experiments, compared with the sham-operation group, the renal tubular injury was more aggravated, Scr and BUN were higher, p62 protein expression was lower, and the protein expression levels of KIM-1, Beclin1 and LC3 were higher in the IR group (all P<0.05). Compared with Foxk1 flox/flox IR goup, renal tubular injury was more alleviated, Scr, BUN and the protein expression levels of KIM-1 and p62 were lower, while the protein expression levels of Beclin1 and LC3 were higher in Foxk1 cKO IR group (all P<0.05). Compared with shCtrl HR group, shFoxk1 HR group had lower protein expression levels of KIM-1 and p62 and higher expression levels of Beclin1 and LC3 in vitro (all P<0.05). Conclusions:The expression of FOXK1 is decreased in ischemic AKI model. Foxk1 deficiency alleviates renal tubular epithelial cell injury and protects against ischemic AKI through activating autophagy.
