Effect and Mechanism of miR-4321 Inhibition Alleviates Sepsis Associated Acute Kidney Injury
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2022.0608
- VernacularTitle:抑制miR-4321减轻脓毒症相关急性肾脏损伤的作用和机制研究
- Author:
Chun-min ZHANG
1
;
Fei-yan CHEN
1
;
Wen-min YANG
1
;
Yong-min LIN
1
;
Bi-tao MAO
1
;
Jie CHEN
1
;
Zhi-yuan WU
1
;
Yu-feng LIANG
1
Author Information
1. Pediatric Intensive Care Unit， Guangzhou Women and Children's Medical Center， Guangzhou Medical University， Guangzhou 510120， China
- Publication Type:Journal Article
- Keywords:
sepsis associated acute kidney injury;
lipopolysaccharide;
miR-4321;
mTOR
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2022;43(6):928-937
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveSepsis associated acute kidney injury （SA-AKI） is a critical clinical disease. The purpose of this study was to investigate the role and molecular mechanism of miR-4321 in the HK2 cellular damage induced by lipopolysaccharides （LPS）. MethodsRT-qPCR was conducted to detect the expression of miR-3165， miR-4270 and miR-4321 in LPS-induced HK2 cell model and cecal ligation and puncture （CLP）-induced renal injury model. In LPS-induced HK2 cell model， miR-4321-inhibitor was used to inhibit the miR-4321 expression. The effects of miR-4321 on cell proliferation， cell viability， cytokines and apoptosis-related protein levels were evaluated by CCK-8 analysis， EdU staining analysis and western blotting analysis， respectively. In CLP-induced renal injury model， miR-4321-Antago was used to intervene the miR-4321 expression. The changes of renal tissue structure were examined by H&E staining. The levels of serum creatinine and blood urea nitrogen （BUN） were measured by colorimetric method. ELISA was employed to assess the expression of interleukin-6 （IL-6）， interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in serum. IHC staining and western blotting were performed to determine the mTOR level and apoptosis-related protein expression in kidney tissues. Target genes of miR-4321 were predicted by Targetscan software and verified by dual-luciferase reporter assay. ResultsCompared with the control group， the miR-4321 expression was increased in LPS-induced HK2 cell model （n=3， t=7.154， P=0.001 3）. miR-4321 inhibitor promoted the proliferation and viability of HK2 cells， decreased the expression of LPS-induced IL-6， IL-1β and TNF-α and apoptosis-related proteins. In vivo experiments showed that miR-4321-Antago inhibited serum creatinine and BUN levels in CLP mice， improved renal injury， reduced levels of IL-1β， IL-6 and TNF-α， promoted the mTOR expression in renal tissues and inhibited the apoptosis-related protein expression. mTOR signaling pathway was believed the target gene of miR-4321. ConclusionInhibition of miR-4321 significantly alleviates SA-AKI， which may be achieved by increasing the expression of mTOR.
