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Preparing a Mice Model of Severe Acute Pancreatitis via a Combination of Caerulein and Lipopolysaccharide Intraperitoneal Injection
Published on: May 10, 2024
MiR-325-3p Alleviates Acute Pancreatitis via Targeting RIPK3
Ao Jia1, Zhen-Wei Yang1, Ji-Yu Shi1
1Tianjin Medical University, Tianjin, 300070, China.
Background And Aims:
Acute pancreatitis (AP) is an acute inflammatory disease that can lead to death. Mir-325-3p is strongly and abnormally expressed in many diseases, necessitating exploration of its function and mechanism in AP.
Methods:
Blood samples from AP patients and mice were analyzed. The expression levels of miR-325-3p in AP patients and mouse were detected. Whether miR-325-3p targets RIPK3 gene was predicted by TargetScan online database and dual luciferase reporter assay. In vitro experiments verified the effect of miR-325-3p overexpression on caerulein-induced MPC83 pancreatic acinar cancer cell line. In vivo experiments verified the effect of overexpression of miR-325-3p on the disease degree of pancreatic tissues in AP mice.
Results:
Analysis of blood samples from AP patients and experiments in mice demonstrated that expression of miR-325-3p was significantly reduced during the process of AP in humans and mice. Predicted using the TargetScan online database and through dual luciferase reporter assay detection, miR-325-3p directly targets the RIPK3 gene. In vitro experiments revealed that overexpression of miR-325-3p reversed caerulein-induced apoptosis and necroptosis in MPC83 pancreatic acinar cancer cell line. We used Z-VAD-FMK to assess necroptosis and demonstrated that miR-325-3p targets necroptosis to reduce cell damage. In subsequent experiments in mice, we verified that overexpression of miR-325-3p reduces inflammation, edema, hemorrhage, and necrosis in acute pancreatitis. Characteristic western blot, immunohistochemistry, and transmission electron microscopy results revealed that overexpression of miR-325-3p reduces the severity of acute pancreatitis by inhibiting pancreatic necroptosis in AP mice.
Conclusions:
The current research results indicate that miR-325-3p directly targets RIPK3 and exerts a protective role in mouse AP. Necroptosis is still the primary mechanism of RIPK3 regulation. MiR-325-3p inhibits acute pancreatitis by targeting RIPK3-dependent necroptosis, which may represent a novel treatment method for acute pancreatitis.
Insights
MicroRNA-325-3p (miR-325-3p) is reduced in acute pancreatitis (AP). Overexpressing miR-325-3p inhibits RIPK3-dependent necroptosis, offering a potential new treatment for AP.
Area of Science:
- Biochemistry
- Molecular Biology
- Pathology
Background:
- Acute pancreatitis (AP) is a severe inflammatory condition with high mortality.
- MicroRNA-325-3p (miR-325-3p) shows abnormal expression in various diseases, prompting investigation into its role in AP.
Purpose of the Study:
- To investigate the function and mechanism of miR-325-3p in acute pancreatitis.
- To determine if miR-325-3p targets the RIPK3 gene and its effect on necroptosis in AP.
Main Methods:
- Analyzed blood samples from AP patients and mice to measure miR-325-3p expression.
- Utilized TargetScan and dual luciferase reporter assays to predict and confirm miR-325-3p targeting of RIPK3.
- Conducted in vitro studies on MPC83 cells and in vivo studies on AP mouse models to assess the effects of miR-325-3p overexpression.
Main Results:
- miR-325-3p expression was significantly decreased in both human and mouse AP.
- miR-325-3p was confirmed to directly target the RIPK3 gene.
- Overexpression of miR-325-3p reduced apoptosis and necroptosis in vitro and alleviated inflammation, edema, hemorrhage, and necrosis in AP mice by inhibiting RIPK3-dependent necroptosis.
Conclusions:
- miR-325-3p plays a protective role in AP by directly targeting RIPK3.
- Inhibition of RIPK3-dependent necroptosis by miR-325-3p presents a potential novel therapeutic strategy for acute pancreatitis.
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