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A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model
Published on: September 11, 2020
MicroRNA-214-5p aggravates sepsis-related acute kidney injury in mice
Cheng Guo1, Fang-Xiong Ye1, Yong-Hong Jian1
1Department of Nephrology, Renmin Hospital of Wuhan University, Wuhan, China.
Abstract:
Acute kidney injury (AKI) is a devastating comorbidity in sepsis and correlates with a very poor prognosis and increased mortality. Currently, we use lipopolysaccharide (LPS) to establish sepsis-related AKI and try to demonstrate the pathophysiological role of microRNA-214-5p (miR-214-5p) in this process. Mice were intravenously injected with the miR-214-5p agomir, antagomir or negative controls for three consecutive days and then received a single intraperitoneal injection of LPS (10 mg/kg) for 24 h to induce AKI. Besides, the Boston University mouse proximal tubular cell lines were stimulated with LPS (10 μg/ml) for 8 h to investigate the role of miR-214-5p in vitro. To inhibit adenosine monophosphate-activated protein kinase (AMPK), compound C (CpC) was used in vivo. For glucagon-like peptide-1 receptor (GLP-1R) silence, cells were transfected with the small interfering RNA against GLP-1R. miR-214-5p level was upregulated in LPS-treated kidneys and proximal tubular cell lines. The miR-214-5p antagomir reduced LPS-induced renal inflammation and oxidative stress, thereby preventing renal damage and dysfunction. In contrast, the miR-214-5p agomir aggravated LPS-induced inflammation, oxidative stress and AKI in vivo and in vitro. Mechanistically, we found that the miR-214-5p antagomir prevented septic AKI via activating AMPK and that CpC treatment completely abrogated its renoprotective effect in mice. Further detection showed that miR-214-5p directly bound to the 3'-untranslational region of GLP-1R to inhibit GLP-1R/AMPK axis. Our data identify miR-214-5p as a promising therapeutic candidate to treat sepsis-related AKI.
Insights
MicroRNA-214-5p (miR-214-5p) is upregulated in sepsis-induced acute kidney injury (AKI). Inhibiting miR-214-5p protects kidneys by activating the GLP-1R/AMPK pathway, offering a potential therapeutic strategy for sepsis-related AKI.
Area of Science:
- Nephrology
- Molecular Biology
- Pharmacology
Background:
- Sepsis-induced acute kidney injury (AKI) is a critical condition with high mortality.
- MicroRNAs play significant roles in the pathogenesis of various diseases, including AKI.
- The specific role of microRNA-214-5p (miR-214-5p) in sepsis-related AKI requires further elucidation.
Purpose of the Study:
- To investigate the role of miR-214-5p in the development of sepsis-related AKI.
- To explore the underlying molecular mechanisms involving miR-214-5p, adenosine monophosphate-activated protein kinase (AMPK), and glucagon-like peptide-1 receptor (GLP-1R).
- To evaluate the therapeutic potential of targeting miR-214-5p for sepsis-related AKI.
Main Methods:
- Sepsis-related AKI was induced in mice using lipopolysaccharide (LPS).
- miR-214-5p agomir and antagomir were administered to mice, and proximal tubular cells were treated with LPS.
- The effects of miR-214-5p modulation on renal inflammation, oxidative stress, and kidney function were assessed.
- Inhibitors of AMPK (compound C) and GLP-1R (siRNA) were used to investigate the molecular pathways.
Main Results:
- miR-214-5p levels were significantly upregulated in LPS-induced AKI in vivo and in vitro.
- Administration of miR-214-5p antagomir attenuated renal inflammation, oxidative stress, and improved kidney function.
- Conversely, miR-214-5p agomir exacerbated LPS-induced kidney damage.
- miR-214-5p inhibition protected against AKI by activating the AMPK pathway, which was dependent on GLP-1R.
Conclusions:
- miR-214-5p is a key mediator in the pathogenesis of sepsis-related AKI.
- Targeting miR-214-5p, potentially through its interaction with the GLP-1R/AMPK axis, represents a promising therapeutic strategy for sepsis-related AKI.
- Further research into miR-214-5p-based therapies could lead to improved patient outcomes in sepsis.
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