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NADPH Oxidase-Mediated Testicular Oxidative Imbalance Regulates the TXNIP/NLRP3 Inflammasome Axis Activation after
Duaah Almarzouq1, May Al-Maghrebi1
1Department of Biochemistry, College of Medicine, Kuwait University, Safat 13110, Kuwait.
Abstract:
Oxidative stress, inflammation and germ cell death are the main characteristics of testicular ischemia reperfusion injury (tIRI), which is considered as the underlying mechanism for testicular torsion and detorsion. The study aimed to examine the effect of tIRI-activated NADPH oxidase (NOX) on the expression of the NLRP3 inflammasome pathway components. Three groups of male Sprague-Dawley rats (n = 12 each) were studied: sham, unilateral tIRI only and tIRI treated with apocynin, a NOX-specific inhibitor. The tIRI rat model was subjected to 1 h of ischemia followed by 4 h of reperfusion. H&E staining, real time PCR, biochemical assays, and Western blot were utilized to evaluate spermatogenic damage, gene expression, oxidative stress markers, and NLRP3 pathway components, respectively. As a result of tIRI, decreased total antioxidant capacity and suppressed activities of superoxide dismutase and catalase were associated with spermatogenic arrest. The components of the NLRP3 inflammasome pathway (TXNIP, NLRP3, ASC, caspase-1, GSDMD, MMP-9) were upregulated transcriptionally and post-transcriptionally during tIRI. In parallel, tissue inflammation was demonstrated by a marked increase in the concentrations of myeloperoxidase, IL-1β, and IL-18. Apocynin treatment prevented testicular oxidative stress and inflammation. Thus, NOX inhibition by apocynin prevented ROS accumulation, proinflammatory cytokine overexpression and NLRP3 inflammasome activation during tIRI.
Insights
Testicular ischemia reperfusion injury (tIRI) activates NADPH oxidase (NOX), leading to oxidative stress and inflammation. Inhibiting NOX with apocynin prevents these effects, protecting against tIRI-induced damage.
Area of Science:
- Reproductive biology
- Pathology
- Molecular biology
Background:
- Testicular ischemia reperfusion injury (tIRI) involves oxidative stress, inflammation, and germ cell death.
- tIRI is a key factor in testicular torsion and detorsion.
- The role of NADPH oxidase (NOX) in tIRI-induced NLRP3 inflammasome activation requires further investigation.
Purpose of the Study:
- To investigate the effect of tIRI-activated NOX on NLRP3 inflammasome pathway components.
- To evaluate the protective effects of apocynin, a NOX inhibitor, against tIRI.
Main Methods:
- Male Sprague-Dawley rats were subjected to unilateral tIRI (1h ischemia, 4h reperfusion) or sham procedure.
- Apocynin was administered to inhibit NOX.
- Assays included H&E staining, real-time PCR, biochemical analyses, and Western blot.
Main Results:
- tIRI led to decreased antioxidant capacity, suppressed superoxide dismutase and catalase activity, and spermatogenic arrest.
- Expression of NLRP3 inflammasome components (TXNIP, NLRP3, ASC, caspase-1, GSDMD, MMP-9) was upregulated.
- Apocynin treatment attenuated oxidative stress, inflammation (myeloperoxidase, IL-1β, IL-18), and NLRP3 inflammasome activation.
Conclusions:
- NOX activation is a critical mediator of oxidative stress and inflammation in tIRI.
- Inhibition of NOX by apocynin ameliorates tIRI-induced testicular damage.
- Targeting NOX represents a potential therapeutic strategy for tIRI.
