Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis
Ischemia–reperfusion injury (IRI) is a predominant factor leading to delayed graft function (DGF) following kidney transplantation. MicroRNAs (miRNAs) play a pivotal role in the pathogenesis of renal IRI, with ferroptosis being a critical driving force throughout the process. In this study, we utili...
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MDPI AG
2024-10-01
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| author | Haitao Yu Xin Gou |
| author_facet | Haitao Yu Xin Gou |
| author_sort | Haitao Yu |
| collection | DOAJ |
| description | Ischemia–reperfusion injury (IRI) is a predominant factor leading to delayed graft function (DGF) following kidney transplantation. MicroRNAs (miRNAs) play a pivotal role in the pathogenesis of renal IRI, with ferroptosis being a critical driving force throughout the process. In this study, we utilized bioinformatics methods to construct a network diagram of differentially expressed miRNAs, transcription factors (TFs), and ferroptosis-related genes. An I/R-induced renal injury model in mice and an in vitro H/R-induced HK-2 cell injury model were established. Quantitative real-time PCR (qRT-PCR) and Western blot analysis were used to measure the mRNA and miRNA levels in cells and tissues. The MDA concentration, iron levels, and GSH concentration were measured to evaluate the ferroptosis levels. CCK-8 assays were performed to assess cell viability. Luciferase reporter assays were conducted to validate the downstream targets of miRNA, and chromatin immunoprecipitation assays were performed to verify the interaction between TFs and mRNAs. Both the in vivo and in vitro results demonstrate that miR-451a was significantly enriched in the IRI renal tissues and cells, exacerbating ferroptosis. MiR-451a was found to reduce the expression of Kruppel-like factor 1 (KLF1) by directly binding to the 3′UTR of KLF1 mRNA. Additionally, KLF1 was identified as a negative transcription factor for acyl-CoA synthetase long-chain family member 4 (ACSL4). We demonstrated that IRI induced the upregulation of miR-451a, which reduced KLF1 expression, thereby promoting ferroptosis by upregulating ACSL4 expression, ultimately aggravating IRI-induced renal damage. |
| format | Article |
| id | doaj-art-c08345dcf63a4847ac2ce4d13faec586 |
| institution | OA Journals |
| issn | 1467-3037 1467-3045 |
| language | English |
| publishDate | 2024-10-01 |
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| spelling | doaj-art-c08345dcf63a4847ac2ce4d13faec5862025-08-20T02:28:12ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452024-10-014611118531186710.3390/cimb46110704Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote FerroptosisHaitao Yu0Xin Gou1Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaIschemia–reperfusion injury (IRI) is a predominant factor leading to delayed graft function (DGF) following kidney transplantation. MicroRNAs (miRNAs) play a pivotal role in the pathogenesis of renal IRI, with ferroptosis being a critical driving force throughout the process. In this study, we utilized bioinformatics methods to construct a network diagram of differentially expressed miRNAs, transcription factors (TFs), and ferroptosis-related genes. An I/R-induced renal injury model in mice and an in vitro H/R-induced HK-2 cell injury model were established. Quantitative real-time PCR (qRT-PCR) and Western blot analysis were used to measure the mRNA and miRNA levels in cells and tissues. The MDA concentration, iron levels, and GSH concentration were measured to evaluate the ferroptosis levels. CCK-8 assays were performed to assess cell viability. Luciferase reporter assays were conducted to validate the downstream targets of miRNA, and chromatin immunoprecipitation assays were performed to verify the interaction between TFs and mRNAs. Both the in vivo and in vitro results demonstrate that miR-451a was significantly enriched in the IRI renal tissues and cells, exacerbating ferroptosis. MiR-451a was found to reduce the expression of Kruppel-like factor 1 (KLF1) by directly binding to the 3′UTR of KLF1 mRNA. Additionally, KLF1 was identified as a negative transcription factor for acyl-CoA synthetase long-chain family member 4 (ACSL4). We demonstrated that IRI induced the upregulation of miR-451a, which reduced KLF1 expression, thereby promoting ferroptosis by upregulating ACSL4 expression, ultimately aggravating IRI-induced renal damage.https://www.mdpi.com/1467-3045/46/11/704ischemia–reperfusion injurymicroRNAferroptosisKruppel-like factor 1acyl-CoA synthetase long-chain family member 4 |
| spellingShingle | Haitao Yu Xin Gou Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis Current Issues in Molecular Biology ischemia–reperfusion injury microRNA ferroptosis Kruppel-like factor 1 acyl-CoA synthetase long-chain family member 4 |
| title | Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis |
| title_full | Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis |
| title_fullStr | Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis |
| title_full_unstemmed | Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis |
| title_short | Overexpression of miR-451a Aggravates Renal Ischemia–Reperfusion Injury by Targeting KLF1-ACSL4 to Promote Ferroptosis |
| title_sort | overexpression of mir 451a aggravates renal ischemia reperfusion injury by targeting klf1 acsl4 to promote ferroptosis |
| topic | ischemia–reperfusion injury microRNA ferroptosis Kruppel-like factor 1 acyl-CoA synthetase long-chain family member 4 |
| url | https://www.mdpi.com/1467-3045/46/11/704 |
| work_keys_str_mv | AT haitaoyu overexpressionofmir451aaggravatesrenalischemiareperfusioninjurybytargetingklf1acsl4topromoteferroptosis AT xingou overexpressionofmir451aaggravatesrenalischemiareperfusioninjurybytargetingklf1acsl4topromoteferroptosis |