Tubuloside A mitigates sepsis-induced splenic injury in mice by suppressing NOX4-associated oxidative stress, inflammation, apoptosis, and mitochondrial dysfunction.
Tubuloside A shows potential in reducing sepsis-induced splenic injury in mice by targeting oxidative stress and inflammation, but human studies are needed to confirm its clinical relevance.
Where it sits
this study against the rest of the mots-c corpusSummary and findings
The study investigated the effects of Tubuloside A (TA) on sepsis-induced splenic injury in mice. TA was found to alleviate splenic injury by reducing oxidative stress, inflammation, apoptosis, and mitochondrial dysfunction. The study utilized a murine cecal ligation and puncture model and LPS-stimulated macrophages to explore these effects.
Abstract
<h4>Ethnopharmacological relevance</h4>Cistanche deserticola Y.C.Ma is a well-known traditional medicinal herb widely used in traditional Chinese medicine for the treatment of kidney injury-related conditions, fatigue, infertility, and age-related disorders, as well as for improving immune function, and is traditionally prescribed for conditions associated with weakness and chronic inflammatory states.<h4>Aim of the study</h4>To investigate the potential efficacy of Tubuloside A (TA), an active constituent of Cistanche deserticola Y.C.Ma, against sepsis-induced splenic injury, a sepsis-associated structural and functional impairment of the spleen characterized by disrupted splenic architecture, dysregulated immune-cell homeostasis, excessive inflammatory responses, and weakened host defense, and to clarify its underlying mechanism of action.<h4>Materials and methods</h4>A murine cecal ligation and puncture (CLP) model and lipopolysaccharide (LPS)-stimulated J774A.1 macrophages were used to investigate the protective effects of TA against sepsis-induced splenic injury. Oxidative stress, antioxidant capacity, mitochondrial function, inflammatory responses, and apoptosis-related injury, splenic immune-cell composition, macrophage inflammatory phenotype, and F4/80/NOX4 colocalization were evaluated by biochemical assays, JC-1 staining, qPCR, Western blotting, flow cytometry, double immunofluorescence staining, and immunohistochemical analyses. Bone marrow-derived macrophages (BMDMs) were further used for supportive validation of macrophage-related inflammatory responses and NOX4 expression. Integrative network pharmacology and molecular docking were employed to identify candidate molecules potentially associated with TA-mediated protection, and NADPH oxidase 4 (NOX4) overexpression was used to further examine the involvement of NOX4-associated oxidative stress in vitro.<h4>Results</h4>TA significantly alleviated splenic injury and improved survival in septic mice. TA reduced oxidative stress, as evidenced by decreased malondialdehyde and reactive oxygen species (ROS) levels and enhanced antioxidant defenses, including superoxide dismutase, catalase, glutathione, and total antioxidant capacity. TA restored mitochondrial membrane potential and improved mitochondrial homeostasis, accompanied by increased TOM20, GPX4, and PGC-1α expression and reduced Drp1 expression. In addition, TA suppressed pro-inflammatory mediators, including TNF-α, IL-1β, IL-6, and iNOS, increased anti-inflammatory IL-10 expression, and reduced Bax and cleaved caspase-3/9 levels, indicating inhibition of apoptosis-related injury. Flow cytometry and BMDM validation further showed that TA regulated splenic immune-cell alterations and macrophage inflammatory responses, while F4/80/NOX4 double immunofluorescence staining indicated that NOX4 expression was associated with F4/80-positive macrophages in splenic tissues. Mechanistically, network pharmacology and molecular docking suggested that NOX4-associated oxidative stress may be involved in TA-mediated protection, which was further supported by the marked induction of NOX4 during sepsis and by the finding that NOX4 overexpression significantly blunted the protective effects of TA in vitro.<h4>Conclusion</h4>This study demonstrates that TA exerts a protective effect against sepsis-induced splenic injury by suppressing NOX4-associated oxidative stress, preserving mitochondrial homeostasis, and limiting downstream inflammatory and apoptotic damage. These findings not only expand the pharmacological profile of TA, but also provide experimental support for the therapeutic potential of an active constituent from Cistanche deserticola Y.C.Ma in sepsis-related immune-organ injury, particularly through the regulation of oxidative stress, macrophage-associated inflammatory responses, and splenic immune-cell alterations.
Background
Sepsis-induced splenic injury is a significant complication characterized by disrupted splenic architecture and immune dysfunction. Traditional Chinese medicine has long utilized Cistanche deserticola for its potential health benefits, including immune modulation. This study aims to explore the efficacy of Tubuloside A, a component of this herb, in mitigating sepsis-related splenic damage, addressing a gap in understanding its mechanistic role in oxidative stress and inflammation.
Methods
The study employed a murine cecal ligation and puncture (CLP) model and LPS-stimulated J774A.1 macrophages to assess the effects of Tubuloside A. Various assays, including biochemical, qPCR, Western blotting, and flow cytometry, were used to evaluate oxidative stress, mitochondrial function, and inflammatory responses. The study also utilized network pharmacology and molecular docking to explore potential molecular interactions.
Results
Tubuloside A significantly reduced oxidative stress markers such as malondialdehyde and ROS levels in septic mice. It enhanced antioxidant defenses and restored mitochondrial membrane potential, as indicated by increased expression of TOM20, GPX4, and PGC-1α. TA also suppressed inflammatory mediators and apoptosis markers, while increasing anti-inflammatory cytokine IL-10. NOX4-associated oxidative stress was implicated in TA's protective effects, as NOX4 overexpression blunted these benefits in vitro.
Interpretation
The findings suggest that Tubuloside A has a protective role against sepsis-induced splenic injury in mice, primarily through the modulation of oxidative stress and inflammatory pathways. While the results are promising, the clinical significance remains uncertain due to the preclinical nature of the study. Further research in human models is necessary to determine its therapeutic potential.
Key findings
- TA significantly alleviated splenic injury and improved survival in septic mice.
- TA reduced oxidative stress, as evidenced by decreased malondialdehyde and ROS levels.
- TA enhanced antioxidant defenses, including superoxide dismutase, catalase, and glutathione.
- TA suppressed pro-inflammatory mediators like TNF-α, IL-1β, IL-6, and iNOS.
- TA increased anti-inflammatory IL-10 expression and reduced Bax and cleaved caspase-3/9 levels.
Limitations
- animal model only, no human data
- surrogate endpoints used
- NOX4 overexpression effects not fully explored
- mechanistic focus limits direct clinical applicability