Ouabain Decreases Regulatory T Cell Number in Mice by Reducing IL-2 Secretion

Silva, Joyle Moreira Carvalho da; Azevedo, Augusto das Neves; Barbosa, Rebeca Pinheiro dos Santos; Teixeira, Mariana Pires; Vianna, Thais Andressa Gonçalves; Fittipaldi, Juliana; Cabral, Vinicius Ribeiro; Paiva, Luciana Souza de

doi:10.1159/000501720

Cited by 6 publications

(4 citation statements)

References 45 publications

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“…The results of the Annexin V/PI investigation suggest that the majority of SK-BR-3 cells treated with cardiotonic glycosides do not undergo a process of necrosis, but die by apoptosis. These results support the data obtained by Da Silva et al [ 26 ] on murine Tregs lymphocytes, by Chang et al [ 10 ] on DU-145 cells, and Khajah et al [ 11 ] on triple-negative breast cancer cells. At micromolar concentrations, the most likely mechanism of ouabain-induced programmed cell death is due to energy substrate depletion.…”

Section: Discussionsupporting

confidence: 91%

In Vitro Study of the Multimodal Effect of Na+/K+ ATPase Blocker Ouabain on the Tumor Microenvironment and Malignant Cells

Harich,

Gavriliuc,

Ordodi

et al. 2023

Biomedicines

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Na+/K+ ATPase is a protein involved in the active transport of ions across the cellular membrane. Ouabain is a cardiotonic glycoside that, by inhibiting the Na+/K+ pump, interferes with cell processes mediated directly by the pump, but also indirectly influences other cellular processes such as cell cycle and proliferation, growth, cell differentiation, angiogenesis, migration, adhesion, and invasion. We used the SK-BR-3 breast cancer cell line, mesenchymal stem cells (MSCs), and tumor-associated fibroblasts (TAFs) in vitro to determine the effects of ouabain exposure on these cellular types. The results showed a multi-level effect of ouabain mainly on tumor cells, in a dose-dependent manner, while the TAFs and their normal counterparts were not significantly influenced. Following exposure to ouabain, the SK-BR-3 cells changed their morphologic appearance, decreased the expression of immunophenotypic markers (CD29, Her2, VEGF), the proliferation rate was significantly decreased (Ki67 index), the cells were blocked in the G0 phase of the cell cycle and suffered necrosis. These data were correlated with the variable expression of α and β Na+/K+ pump subunits in tumor cells, resulting in decreased ability to adhere to the VCAM-1 substrate in functional flow chamber studies. Being indicative of the pro-apoptotic and inhibitory effect of ouabain on tumor invasion and metastasis, the results support the addition of ouabain to the oncological therapeutic arsenal, trailing the “repurposing drugs” approach.

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Section: Discussionsupporting

confidence: 91%

In Vitro Study of the Multimodal Effect of Na+/K+ ATPase Blocker Ouabain on the Tumor Microenvironment and Malignant Cells

Harich,

Gavriliuc,

Ordodi

et al. 2023

Biomedicines

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“…In addition, digoxin decreased the number of Th17 cells and increased the number of T-Regs, which promoted immunosuppression in mice [109]. Ouabain has been shown to reduce the number of T-regs by decreasing Il-2 production by T lymphocytes [107]. The regulation of lymphocytes by CTS will offer new and modified treatment strategies for rheumatologic and autonomic diseases such as Chagrin syndrome [90], rheumatoid arthritis [109] via promoting T-Regs and suppressing joint inflammation and bone erosion in mice CIA (collagen induced arthritis) arthritis model.…”

Section: Regulatory T-cellsmentioning

confidence: 99%

“…Telocinobufagin has also been shown to dramatically enhance the proliferative response of splenocytes to concanavalin A, ovalbumin and substantially increase transcription factors T-bet (Th1) mRNA level and the CD3(+)/CD3(+)CD4(+)/CD3(+)CD8(+) phenotype in splenocytes from ovalbumin-immunized mice [106]. There are contradictory data on the effect of ouabain on the number of T-helpers cells, for example, in a study [107] it was shown that intraperitoneal injection of ouabain in mice significantly decreased the number of CD4+ T-lymphocytes in the spleen, especially regulatory T-cells, while the percentage and total number of lymphocytes in mesenteric lymph nodes remained the same [108]. Reduction of levels of antigen-specific T-helper lymphocytes cytokines (IL-4, IL-5, IL-13) in bronchoalveolar lavage fluid by bufalin was described above [104].…”

Section: T-helpersmentioning

confidence: 99%

Mechanisms providing effects of cardiotonic steroids in mammalian cells with special attention to blood cells

Poluektov,

Lopina,

Strelkova

et al. 2023

Preprint

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Cardiotonic steroids (CTS) are a group of steroid compounds derived from certain plants and animals. CTS are selective inhibitors of Na,K-ATPase, so for a long time they were widely used for medical purposes to treat heart failure and some other diseases. However, over time, doctors gradually began to refuse this group of drugs due to their narrow therapeutic range and a number of serious side effects. However, in parallel with the rise and fall of interest in CTS as a drug, interest in them as a single class of high-affinity Na,K-ATPase inhibitors has only increased. Numerous data on the effect of CTS on signaling cascades and cell viability made impelling a search for a rationale for the existence of such a subtle biological regulator. A large number of studies devoted to the potential use of CTS in the treatment of neurodegenerative, oncological, immune and other diseases "revitalized" the already "written off" group of the drugs. If initially the role of exogenous (mainly plant origin) CTS was studied, then by the 90s of the XX century the study of the role of endogenous CTS and their regulatory effects became a popular trend in biology and physiology. The role of endogenous CTS was studied in details in the pathogenesis of cardiovascular diseases [1]. A large number of researchers have unanimously stated that endogenous CTS are very complex regulators of the development of pathological processes; however, the exact cause-and-effect relationship that would unambiguously explain the processes of synthesis, utilization, and the role of endogenous CTS has not yet been established. The effects of CTS in the frame of various regulatory schemes have been described, but the predominant number of studies was devoted to arterial hypertension, heart and renal failure, cancer [2–4], aging [5,6], and neuroinflammation. The effects of both endo- and exogenous CTS on blood cells and have been studied to a much lesser extent.

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“…Besides, ouabain suppressed human renal cancer OS-RC-2 cell proliferation by targeting to the NKA α3 isoform (32). Furthermore, ouabain reduced IL-2 secretion and resulted in a decrease in regulatory T cell numbers in mice (33).…”

mentioning

confidence: 98%

Ouabain Induces DNA Damage in Human Osteosarcoma U-2 OS Cells and Alters the Expression of DNA Damage and DNA Repair–associated Proteins

Yang

Chen

et al. 2021

In Vivo

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Background/Aim: Ouabain, isolated from natural plants, exhibits anticancer activities; however, no report has presented its mechanism of DNA damage induction in human osteosarcoma cancer cells in vitro. The aim of this study was to investigate whether ouabain induces DNA damage and repair, accompanied with molecular pathways in human osteosarcoma cancer U-2 OS cells in vitro. Materials and Methods: The percentage of viable cell number was measured by flow cytometric assay; DNA damage was assayed by DAPI staining, comet assay, and agarose gel electrophoresis. DNA damage and repair associated protein expressions were assayed by western blotting assays. Results: Ouabain reduced total cell viability, induced chromatin condensation, DNA fragmentation, and DNA damage in U-2 OS cells. Ouabain increased p-ATM Ser1981 , p-ATR Ser428 , and p53 at 2.5-10 μM, increased p-p53 Ser15 at 10 μM; however, it decreased p-MDM2 Ser166 at 2.5-10 μM. Ouabain increased p-H2A.X Ser139 , MDC-1, and PARP at 2.5-10 μM and BRCA1 at 5-10 μM; however, it decreased DNA-PK and MGMT at 2.5-10 μM in U-2 OS cells at 48 h treatment. Ouabain promoted expression and nuclear translocation of p-H2A.X Ser139 in U-2 OS cells and this was confirmed by confocal laser microscopy. Conclusion: Ouabain reduced total viable cell number through triggering DNA damage and altering the protein expression of DNA damage and repair system in U-2 OS cells in vitro. Osteosarcoma (OS) originates from long bones, including the humerus, femur, and tibia (1, 2) and it is the most frequent type of malignant bone cancer. OS contributes to cancer-2687 This article is freely accessible online.

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Ouabain Decreases Regulatory T Cell Number in Mice by Reducing IL-2 Secretion

Cited by 6 publications

References 45 publications

In Vitro Study of the Multimodal Effect of Na+/K+ ATPase Blocker Ouabain on the Tumor Microenvironment and Malignant Cells

In Vitro Study of the Multimodal Effect of Na+/K+ ATPase Blocker Ouabain on the Tumor Microenvironment and Malignant Cells

Mechanisms providing effects of cardiotonic steroids in mammalian cells with special attention to blood cells

Ouabain Induces DNA Damage in Human Osteosarcoma U-2 OS Cells and Alters the Expression of DNA Damage and DNA Repair–associated Proteins

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