Effect of microRNA-138 on the ischemia/reperfusion of human renal tubular epithelial cells
10.3969/j.issn.1673-9701.2024.11.012
- VernacularTitle:微小RNA-138/NGAL对缺血再灌注诱导人肾小管上皮细胞的影响
- Author:
Donghao QIU
1
,
2
Author Information
1. Department of Nephrology, Affiliated Hangzhou First People&rsquo
2. s Hospital, Zhejiang University School of Medicine, Hangzhou 310008, Zhejiang, China
- Publication Type:Journal Article
- Keywords:
Human renal tubular epithelial cells;
Ischemia/reperfusion;
Neutrophil gelatinase related lipid transporter;
TLR4/NF-κ B pathway
- From:
China Modern Doctor
2024;62(11):48-53
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of microRNA-138(miR-138)on injury of ischemia/reperfusion(I/R)induced human renal tubular epithelium(HK-2)cells through neutrophil gelatinase-associated lipocalin(NGAL).Methods HK-2 cells were used to construct I/R model cells,and transfected with miR-138 mimic,miR-138 inhibitor,NGAL,NGAL + miR-138 mimic plasmids,respectively.qRT-PCR determined the expression of miR-138 or NGAL mRNA in different cells to identify the transfection results.Cell counting kit-8(CCK-8)method and flow cytometry were used to detected the activities and apoptosis of cells.ELISA and western blot were used to determine the effects of miR-138 mimic or miR-138 inhibitor on levels of interleukin(IL)-6,IL-1β,tumor necrosis factor(TNF-α)and protein expression of toll like receptor 4(TLR4),nuclear factor kappa-B(NF-κB),inhibitor of NF-κB(IκBα),pho-IκBα(p-IκBα),NGAL of cells.Results miR-138 mRNA expression and cell activity were decreased,while apoptosis increased in I/R cells(P<0.01).Plasmid transfected well,miR-138 mimic increased activity while decreased apoptosis and NGAL mRNA expression of I/R cell.miR-138 inhibitor or NGAL mimic inhibited activity and increased apoptosis and NGAL mRNA expression of I/R cell.The negative effects of NGAL mimic on I/R cell were reversed by miR-138 mimic.miR-138 inhibitor increased levels of IL-6,IL-1β,TNF-α of I/R cell,and increased TLR4,NF-κB,p-IκBα,NGAL protein expression and decreased IκBα protein expression(P<0.05).While miR-138 mimic decreased levels of IL-6,IL-1β,TNF-α of I/R cell,and decreased TLR4,NF-κB,p-IκBα,NGAL protein expression and increased IκBα protein expression(P<0.05).Conclusion miR-138 reduced apoptosis and inflammation factor levels to play a protective role on I/R induced HK-2 cells may through regulating NGAL and TLR4/NF-κB pathway.
