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Use of a Hanging-weight System for Liver Ischemia in Mice
Published on: August 7, 2012
miR‑140‑5p alleviates mouse liver ischemia/reperfusion injury by targeting CAPN1
Qiwen Yu1, Sanyang Chen2, Hongwei Tang1
1Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Abstract:
Ischemia/reperfusion (I/R)‑induced liver injury remains a primary concern in liver transplantation and hepatectomy. Previous studies have indicated that microRNAs (miRs) are involved in multiple pathophysiological processes, including liver I/R. miR‑140‑5p reportedly inhibits inflammatory responses and apoptosis in several diseases; however, the role of miR‑140‑5p in liver I/R remains unknown. The present study aimed to investigate the potential role and mechanism of miR‑140‑5p on liver I/R injury. Mouse liver I/R and mouse AML12 cell hypoxia/reoxygenation (H/R) models were established. miR‑140‑5p mimics, inhibitor or agonists were used to overexpress or inhibit miR‑140‑5p in vitro and in vivo. Reverse transcription‑quantitative polymerase chain reaction was used to detect miR‑140‑5p expression. Liver and cell injury were evaluated using several biochemical assays. The association between miR‑140‑5p and calpain‑1 (CAPN1) was confirmed using a dual‑luciferase reporter assay. The results revealed that miR‑140‑5p expression was decreased in the mouse model of liver I/R injury and AML12 cells subjected to H/R, while overexpressed miR‑140‑5p reduced liver injury in vivo and cell injury in vitro. In addition, CAPN1 was determined to be a target of miR‑140‑5p; overexpressed CAPN1 abrogated the effect of miR‑140‑5p on H/R‑induced cell injury. The present study indicated that miR‑140‑5p protected against liver I/R by targeting CAPN1, which may provide a novel therapeutic target for liver I/R injury.
Insights
MicroRNA-140-5p protects against liver ischemia/reperfusion (I/R) injury by targeting calpain-1. This finding offers a potential new therapeutic strategy for liver I/R damage.
Area of Science:
- Molecular Biology
- Hepatology
- Biochemistry
Background:
- Ischemia/reperfusion (I/R)-induced liver injury is a significant clinical challenge in liver transplantation and surgery.
- MicroRNAs (miRs) are implicated in various pathophysiological processes, including liver I/R injury, but the specific role of miR-140-5p is not well understood.
Purpose of the Study:
- To investigate the role and underlying mechanism of miR-140-5p in liver I/R injury.
- To determine if miR-140-5p can serve as a therapeutic target for mitigating liver I/R damage.
Main Methods:
- Established mouse models of liver I/R and AML12 cell hypoxia/reoxygenation (H/R).
- Utilized miR-140-5p mimics and inhibitors for in vitro and in vivo manipulation.
- Quantified miR-140-5p expression using RT-qPCR and assessed liver/cell injury via biochemical assays.
- Confirmed the interaction between miR-140-5p and calpain-1 (CAPN1) using a dual-luciferase reporter assay.
Main Results:
- miR-140-5p expression was significantly downregulated in liver I/R and H/R models.
- Overexpression of miR-140-5p ameliorated liver and cell injury in both in vivo and in vitro settings.
- Calpain-1 (CAPN1) was identified as a direct target of miR-140-5p.
- Upregulation of CAPN1 reversed the protective effects of miR-140-5p against H/R-induced cell injury.
Conclusions:
- miR-140-5p exerts a protective effect against liver I/R injury.
- This protective mechanism involves the targeting of calpain-1 (CAPN1).
- miR-140-5p represents a promising therapeutic target for managing liver I/R injury.

