心脏成纤维细胞(CF)是间充质型细胞,负责维持心脏细胞外基质(ECM)的稳态。它们的功能障碍导致ECM蛋白的过度分泌,组织硬化,营养和氧交换受损,心脏的电异常.此外,CF作为心脏组织微环境中的前哨细胞,对可能影响心脏功能的各种刺激做出反应。有害刺激诱导CF的炎症反应,增加细胞因子如IL-1β和TNF-α的分泌和细胞粘附分子如ICAM1和VCAM1的表达,最初通过募集免疫细胞促进损伤的解决。然而,持续的有害刺激导致慢性炎症过程和心脏功能障碍。因此,有必要研究控制CF炎症的机制。NFκB是心脏炎症过程的关键调节因子,寻找NFκB调节和CF炎症反应的机制对于开发新的心血管疾病治疗方案至关重要。SGK1,一种丝氨酸-苏氨酸蛋白激酶,是NFκB的调节因子之一,参与血管紧张素II和醛固酮的纤维化作用,以及CF分化。然而,其在CF炎症反应中的作用尚不清楚。另一方面,许多生物活性天然产物已经证明了抗炎作用,但它们在CF炎症中的作用尚不清楚。一种这样的分子是波丁,从Boldo(Peumusboldus)获得的生物碱,智利特有的树,具有良好的细胞保护作用。然而,其参与SGK1和CF炎症的调节是未知的。在这项研究中,我们评估了SGK1和boldine在从新生Sprague-Dawley大鼠分离的CF中的炎症反应中的作用。使用特异性SGK1抑制剂GSK650394分析SGK1的参与。我们的结果表明,SGK1对于CF中LPS和IFN-γ诱导的炎症反应至关重要(细胞因子表达,细胞粘附分子表达,和白细胞粘附)。此外,使用条件培养基(经受冷冻/解冻循环的CF的细胞内含量)来模拟无菌炎症条件。条件培养基在CF中诱导了有效的炎症反应,SGK1抑制剂完全阻止了这种情况。最后,我们的结果表明,波定抑制SGK1激活和由LPS诱导的CF炎症反应,IFN-γ,和CF条件培养基。一起来看,我们的结果将SGK1定位为CF炎症反应的重要调节因子,而将波定定位为心血管疾病中的有前途的抗炎药.
Cardiac fibroblasts (CF) are mesenchymal-type cells responsible for maintaining the homeostasis of the heart\'s extracellular matrix (ECM). Their dysfunction leads to excessive secretion of ECM proteins, tissue stiffening, impaired nutrient and oxygen exchange, and electrical abnormalities in the heart. Additionally, CF act as sentinel cells in the cardiac tissue microenvironment, responding to various stimuli that may affect heart function. Deleterious stimuli induce an inflammatory response in CF, increasing the secretion of cytokines such as IL-1β and TNF-α and the expression of cell adhesion molecules like ICAM1 and VCAM1, initially promoting damage resolution by recruiting immune cells. However, constant harmful stimuli lead to a chronic inflammatory process and heart dysfunction. Therefore, it is necessary to study the mechanisms that govern CF inflammation. NFκB is a key regulator of the cardiac inflammatory process, making the search for mechanisms of NFκB regulation and CF inflammatory response crucial for developing new treatment options for cardiovascular diseases.
SGK1, a serine-threonine protein kinase, is one of the regulators of NFκB and is involved in the fibrotic effects of angiotensin II and aldosterone, as well as in CF differentiation. However, its role in the CF inflammatory response is unknown. On the other hand, many bioactive natural products have demonstrated anti-inflammatory effects, but their role in CF inflammation is unknown. One such molecule is boldine, an alkaloid obtained from Boldo (Peumus boldus), a Chilean endemic tree with proven cytoprotective effects. However, its involvement in the regulation of
SGK1 and CF inflammation is unknown. In this study, we evaluated the role of
SGK1 and boldine in the inflammatory response in CF isolated from neonatal Sprague-Dawley rats. The involvement of
SGK1 was analyzed using GSK650394, a specific
SGK1 inhibitor. Our results demonstrate that
SGK1 is crucial for LPS- and IFN-γ-induced inflammatory responses in CF (cytokine expression, cell adhesion molecule expression, and leukocyte adhesion). Furthermore, a conditioned medium (intracellular content of CF subject to freeze/thaw cycles) was used to simulate a sterile inflammation condition. The conditioned medium induced a potent inflammatory response in CF, which was completely prevented by the SGK1 inhibitor. Finally, our results indicate that boldine inhibits both SGK1 activation and the CF inflammatory response induced by LPS, IFN-γ, and CF-conditioned medium. Taken together, our results position SGK1 as an important regulator of the CF inflammatory response and boldine as a promising anti-inflammatory drug in the context of cardiovascular diseases.