Davidson Carolina Bordin, El Sabbagh Dana El Soufi, Machado Amanda Kolinski, Pappis Lauren, Sagrillo Michele Rorato, Somacal Sabrina, Emanuelli Tatiana, Schultz Júlia Vaz, Augusto Pereira da Rocha João, Santos André Flores Dos, Fagan Solange Binotto, Silva Ivana Zanella da, Andreazza Ana Cristina, Machado Alencar Kolinski
Graduate Program in Nanosciences, Franciscan University, Santa Maria 97010-030, RS, Brazil.
Laboratory of Cell Culture and Bioactive Effects, Franciscan University, Santa Maria 97010-030, RS, Brazil.
Biology (Basel). 2024 Sep 17;13(9):729. doi: 10.3390/biology13090729.
Inflammation is a vital mechanism that defends the organism against infections and restores homeostasis. However, when inflammation becomes uncontrolled, it leads to chronic inflammation. The NLRP3 inflammasome is crucial in chronic inflammatory responses and has become a focal point in research for new anti-inflammatory therapies. Flavonoids like catechin, apigenin, and epicatechin are known for their bioactive properties (antioxidant, anti-inflammatory, etc.), but the mechanisms behind their anti-inflammatory actions remain unclear. This study aimed to explore the ability of various flavonoids (isolated and combined) to modulate the NLRP3 inflammasome using in silico and in vitro models. Computer simulations, such as molecular docking, molecular dynamics, and MM/GBSA calculations examined the interactions between bioactive molecules and NLRP3 PYD. THP1 cells were treated with LPS + nigericin to activate NLRP3, followed by flavonoid treatment at different concentrations. THP1-derived macrophages were also treated following NLRP3 activation protocols. The assays included colorimetric, fluorometric, microscopic, and molecular techniques. The results showed that catechin, apigenin, and epicatechin had high binding affinity to NLRP3 PYD, similar to the known NLRP3 inhibitor MCC950. These flavonoids, particularly at 1 µg/mL, 0.1 µg/mL, and 0.01 µg/mL, respectively, significantly reduced LPS + nigericin effects in both cell types and decreased pro-inflammatory cytokine, caspase-1, and NLRP3 gene expression, suggesting their potential as anti-inflammatory agents through NLRP3 modulation.
炎症是一种重要机制，可保护机体免受感染并恢复体内平衡。然而，当炎症变得不受控制时，就会导致慢性炎症。NLRP3炎性小体在慢性炎症反应中至关重要，已成为新型抗炎疗法研究的焦点。儿茶素、芹菜素和表儿茶素等类黄酮以其生物活性特性（抗氧化、抗炎等）而闻名，但其抗炎作用背后的机制仍不清楚。本研究旨在使用计算机模拟和体外模型探索各种类黄酮（分离的和组合的）调节NLRP3炎性小体的能力。计算机模拟，如分子对接、分子动力学和MM/GBSA计算，研究了生物活性分子与NLRP3 PYD之间的相互作用。用LPS+尼日利亚菌素处理THP1细胞以激活NLRP3，然后用不同浓度的类黄酮处理。THP1衍生的巨噬细胞也按照NLRP3激活方案进行处理。检测方法包括比色法、荧光法、显微镜法和分子技术。结果表明，儿茶素、芹菜素和表儿茶素与NLRP3 PYD具有高结合亲和力，类似于已知的NLRP3抑制剂MCC950。这些类黄酮，特别是分别以1μg/mL、0.1μg/mL和0.01μg/mL的浓度，显著降低了两种细胞类型中LPS+尼日利亚菌素的作用，并降低了促炎细胞因子、半胱天冬酶-1和NLRP3基因的表达，表明它们通过调节NLRP3具有作为抗炎剂的潜力。
