Giordano Magalì, Femminò Saveria, Blua Federica, Boccato Francesca, Rubeo Chiara, Mantuano Beatrice, Cioffi Francesca, Comità Stefano, Brovero Arianna, Ciullo Rosa, Bertinaria Massimo, Penna Claudia, Pagliaro Pasquale
Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, Orbassano, TO 10043, Italy.
Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, 10125 Torino, Italy.
Vascul Pharmacol. 2025 Jun;159:107487. doi: 10.1016/j.vph.2025.107487. Epub 2025 Mar 15.
Cardiovascular diseases remain the leading cause of disability and death in the Western world. Effective cardioprotection involves limiting ischemia/reperfusion injury (IRI), including cell death (pyroptosis) driven by the NLRP3 inflammasome. While various cardiac resident cellular populations contribute to cardioprotection, it remains unclear whether targeting resident macrophages is inherently cardioprotective. Given that INF150, an NLRP3 inhibitor, exhibits varying abilities to penetrate cardiomyocytes and macrophages, we sought to address this question.
We studied the cardioprotective potential of INF150, the potent metabolite of the NLRP3 inhibitor INF195, in isolated hearts or cells. In isolated hearts, we measured infarct size, caspase-1 cleavage, and interleukins (IL) release, while in macrophages, naïve H9c2 and differentiated H9c2 cells, we analyzed cell viability, and pyroptosis markers, including IL-1β release and Gasdermin D cleavage, following hypoxia/reoxygenation (H/R).
While INF150 effectively shielded macrophages from LPS/ATP challenges, it failed to penetrate H9c2 and differentiated H9c2, even at high concentrations (no changes in pyroptosis markers induced by H/R). In the isolated mice heart model, INF150 did not demonstrate cardioprotective effects: infarct size, IL-1β, cleaved caspase-1 levels did not change significantly across tested concentrations of INF150. These findings suggest that while INF150 shows promise in macrophage/phagocytic models, its inability to penetrate cardiomyocytes limits its effectiveness in the whole cardiac tissue. Our results underscore the importance of cardiomyocyte uptake for effective cardioprotection, highlighting the need for NLRP3 inhibitors capable of targeting these cells directly. Future research should focus on enhancing the delivery and cardiomyocyte uptake of NLRP3 inhibitors to achieve cardioprotection. Unlike its precursor, INF195, which penetrates H9c2 cells, INF150 does not appear to offer cardioprotection in the whole organ.
心血管疾病仍是西方世界致残和致死的主要原因。有效的心脏保护包括限制缺血/再灌注损伤（IRI），其中包括由NLRP3炎性小体驱动的细胞死亡（焦亡）。虽然各种心脏驻留细胞群都有助于心脏保护，但靶向驻留巨噬细胞本身是否具有心脏保护作用仍不清楚。鉴于NLRP3抑制剂INF150穿透心肌细胞和巨噬细胞的能力各不相同，我们试图解决这个问题。
我们研究了NLRP3抑制剂INF195的有效代谢产物INF150在离体心脏或细胞中的心脏保护潜力。在离体心脏中，我们测量梗死面积、半胱天冬酶-1裂解情况以及白细胞介素（IL）释放，而在巨噬细胞、未分化的H9c2细胞和分化的H9c2细胞中，我们分析了缺氧/复氧（H/R）后的细胞活力以及焦亡标志物，包括IL-1β释放和Gasdermin D裂解情况。
虽然INF150有效地保护巨噬细胞免受LPS/ATP刺激，但即使在高浓度下，它也无法穿透H9c2细胞和分化的H9c2细胞（H/R诱导的焦亡标志物无变化）。在离体小鼠心脏模型中，INF150未表现出心脏保护作用：在测试的INF150浓度范围内，梗死面积、IL-1β、裂解的半胱天冬酶-1水平均无显著变化。这些发现表明，虽然INF150在巨噬细胞/吞噬模型中显示出前景，但其无法穿透心肌细胞限制了其在整个心脏组织中的有效性。我们的结果强调了心肌细胞摄取对有效心脏保护的重要性，突出了需要能够直接靶向这些细胞的NLRP3抑制剂。未来的研究应专注于增强NLRP3抑制剂的递送和心肌细胞摄取以实现心脏保护。与其能够穿透H9c2细胞的前体INF195不同，INF150似乎无法在整个器官中提供心脏保护。
