强心苷源自植物和动物，自古以来就已被人们所认识。这些物质阻碍真核细胞内钠钾泵的功能。许多报告表明这些化合物影响核受体的活性。因此，我们评估了无毒浓度的各种强心苷对 RORγ 和 RORγT 的影响。 RORγT 是参与 Th17 淋巴细胞分化的关键蛋白。分析的 16 种强心苷在 HepG2 细胞中表现出不同的毒性，所有这些都表现出对 RORγ 的激动作用，这在 RORγ-HepG2 报告细胞系中得到了证实。 RORγ 和 RORγT 亚型的过度表达增强了这些化合物的作用。此外，这些苷类还诱导 HepG2 细胞中 G6PC（一种受 RORγ 调节的基因）的表达。随后，在 Th17 原代淋巴细胞中评估了两种内源性强心苷（marinobufagenin 和哇巴因）和三种最有效的苷（蟾蜍灵、夹竹桃苷和 telecinobufagenin）的作用。所有这些化合物均增加了 IL17A、IL17F、IFNG 和 CXCL10 基因的表达，但它们对 GZMB 和 CCL20 表达表现出不同的影响。分子对接分析揭示了强心苷与 RORγ/RORγT 受体配体结合域的强大结合亲和力。因此，我们证明，在无毒浓度下，强心苷对 RORγ/RORγT 核受体具有激动作用，增强其活性。在对抗各种类型癌症的过继疗法中，可以利用这种潜力来调节表达 IL17 的细胞（例如 Th17 或 Tc17 淋巴细胞）的表型。版权所有 © 2024。由 Elsevier Inc. 出版。

Cardiac glycosides, derived from plants and animals, have been recognized since ancient times. These substances hinder the function of the sodium-potassium pump within eukaryotic cells. Many reports have shown that these compounds influence the activity of nuclear receptors. Thus, we assessed the effects of various cardiac glycosides at nontoxic concentrations on RORγ and RORγT. RORγT is a crucial protein involved in the differentiation of Th17 lymphocytes. Sixteen analyzed cardiac glycosides exhibited varying toxicities in HepG2 cells, all of which demonstrated agonistic effects on RORγ, as confirmed in the RORγ-HepG2 reporter cell line. The overexpression of both the RORγ and RORγT isoforms intensified the effects of these compounds. Additionally, these glycosides induced the expression of G6PC, a gene regulated by RORγ, in HepG2 cells. Subsequently, the effects of two endogenous cardiac glycosides (marinobufagenin and ouabain) and the three most potent glycosides (bufalin, oleandrin, and telecinobufagenin) were evaluated in Th17 primary lymphocytes. All of these compounds increased the expression of the IL17A, IL17F, IFNG, and CXCL10 genes, but they exhibited varying effects on GZMB and CCL20 expression. Molecular docking analysis revealed the robust binding affinity of cardiac glycosides for the ligand binding domain of the RORγ/RORγT receptors. Thus, we demonstrated that at nontoxic concentrations, cardiac glycosides have agonistic effects on RORγ/RORγT nuclear receptors, augmenting their activity. This potential can be harnessed to modulate the phenotype of IL17-expressing cells (e.g., Th17 or Tc17 lymphocytes) in adoptive therapy for combating various types of cancer.Copyright © 2024. Published by Elsevier Inc.