Related Experiment Video
Updated: Oct 8, 2025
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
CPNE3 interaction with RACK1 protects against myocardial ischemia/reperfusion injury
Xiaoqun Zhang1, Xue Han1, Yanan Zhang1
1Cardiology Department One, Cangzhou Central Hospital, Cangzhou, Hebei 061001, P.R. China.
Insights
Copine 3 (CPNE3) protects against myocardial ischemia/reperfusion (I/R) injury by interacting with RACK1. CPNE3 upregulation enhances cell viability and reduces inflammation and apoptosis, offering therapeutic potential for I/R patients.
Area of Science:
- Cardiology
- Molecular Biology
- Cell Biology
Background:
- Acute myocardial ischemia/reperfusion (I/R) poses significant clinical challenges.
- Copine 3 (CPNE3) and receptor for activated C kinase 1 (RACK1) are implicated as risk factors in I/R injury.
Purpose of the Study:
- To investigate the role of CPNE3 in myocardial I/R injury.
- To elucidate the interaction between CPNE3 and RACK1 in the context of I/R.
Main Methods:
- Utilized H9c2 cell lines and a rat myocardial I/R model.
- Assessed gene and protein expression via RT-qPCR and Western blotting.
- Evaluated cell viability (CCK-8), LDH release, inflammatory cytokines, and apoptosis (TUNEL staining).
- Confirmed CPNE3-RACK1 interaction using immunoprecipitation assays.
Main Results:
- CPNE3 and RACK1 expression decreased in hypoxia/reoxygenation (H/R)-induced cardiomyocytes and myocardial I/R models.
- CPNE3 overexpression increased RACK1 levels, enhanced cell viability, and reduced LDH release in H/R-induced cells.
- CPNE3 mitigated inflammation and apoptosis in H/R cardiomyocytes by activating RACK1.
Conclusions:
- CPNE3 plays a protective role in myocardial I/R injury through interaction with RACK1.
- CPNE3 activation of RACK1 suppresses inflammatory responses and apoptosis.
- This study provides novel insights into preventing and treating myocardial I/R injury.
Abstract:
Copine 3 (CPNE3) and receptor for activated C kinase 1 (RACK1) have been determined to be risk factors for patients with acute myocardial ischemia/reperfusion (I/R). The present study aimed to evaluate the role of CPNE3 and its interaction with RACK1 in myocardial (I/R) injury. Reverse transcription-quantitative PCR (RT-qPCR) and western blotting were performed to detect CPNE3 and RACK1 expression levels in H9c2 cells before and after the transfection of CPNE3 overexpression plasmid or small interfering RNA-RACK1. Cell viability was detected using a Cell Counting Kit-8 assay, and immunoprecipitation assays were performed to determine the interaction between CPNE3 and RACK1. A commercial kit was used to examine lactate dehydrogenase (LDH) levels. The expression levels of inflammatory cytokines were detected via RT-qPCR and western blotting. Cell apoptosis was assessed via TUNEL staining and western blotting. The results demonstrated that the expression levels of CPNE3 and RACK1 were decreased in hypoxia/reoxygenation (H/R)-induced H9c2 cardiomyocytes, which was consistent with the expression levels observed in the myocardial I/R injury rat model. It was found that CPNE3 overexpression upregulated RACK1 expression, increased cell viability and suppressed the release of LDH in H/R-induced H9c2 cells. Furthermore, CPNE3 overexpression inhibited the release of inflammatory cytokines and decreased cell apoptosis in H/R-induced cardiomyocytes by activating RACK1 expression. The present study suggested that CPNE3 served an important role in preventing I/R injury by interacting with RACK1, providing novel insight into the prevention of myocardial I/R injury, as well as the treatment and care of patients with myocardial I/R.
