Blocking RIPK2 Function Alleviates Myocardial Ischemia/Reperfusion Injury by Regulating the AKT and NF-κB Pathways

ABSTRACT

Objective

Inflammation and oxidation brought on by myocardial ischemia-reperfusion (MI/R) injury lead to cardiomyocyte apoptosis and necrosis. The receptor interacting serine/threonine kinase 2 (RIPK2) plays significant roles in oxidative stress and excessive inflammation. The purpose of this research is to examine the roles of RIPK2 in MI/R injury.

Methods

The in vivo animal model was constructed by acute coronary I/R, and the in vitro cell model was established by oxygen and glucose deprivation/reperfusion (OGD/R)-stimulated cardiomyocyte injury. RIPK2 expression was examined using qRT-PCR and Western blot. CCK-8 was proposed as a method for detecting cell proliferation. ELISA was utilized to measure inflammatory cytokines (TNF-α, IL-6, and IL-1β) and myocardial injury indicators (CK-MB, Mb, cTnI, and LDH). The levels of MDA and ROS were determined by the kit and fluorescent probe. H&E was conducted to assess MI/R injury after silencing of RIPK2.

Results

In MI/R rats and OGD/R-treated H9C2 cardiomyocytes, RIPK2 was overexpressed at both the mRNA and protein levels. RIPK2 inhibition promoted cell proliferation while inhibiting apoptosis, as evidenced by decreased TUNEL-positive cells and cleaved caspase-3. RIPK2 inhibition reduced MDA and ROS levels, as well as the contents of inflammatory factors. RIPK2 silencing reduced CK-MB, Mb, cTnI, and LDH levels in rat serum and alleviated MI/R injury. Furthermore, RIPK2 inhibition increased p-AKT while decreasing NF-B p-p65 expression.

Conclusion

Silencing of RIPK2 reduced apoptosis, proinflammatory factors, and oxidative stress in MI/R by activating AKT and suppressing NF-κB signals, suggesting a potential therapeutic strategy for MI/R injury.

Graphical abstract

The graphical abstract displayed the role of RIPK2 in heart. RIPK2 inhibition promoted cell proliferation and inhibited apoptosis, inflammatory factors, and oxidative stress in OGD/R injury cells or MI/R injury tissues. Meanwhile, it reduced cardial ischemia and improved cardiac function following MI/R injury. RIPK2 inhibition activated AKT while suppressing NF-κB signaling pathways.

KEYWORDS:

• AKT/NF-κB
• myocardial ischemia-reperfusion
• receptor interacting serine/threonine kinase 2

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Guofang Sun designed the study. Zhen Xia performed the experiments and wrote the manuscript. All authors read and approved the submitted version.

Data availability statement

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.

Additional information

Funding

This work was supported by Jiangxi Provincial Natural Science Foundation [20202BABL206010].