Inhibition of BRD4 alleviates pyroptosis induced by testicular ischemia/reperfusion injury through the NLRP3 pathway.

BACKGROUND

Testicular ischemia/reperfusion injury (IRI) represents a significant contributor to male infertility. BRD4, an epigenetic regulator, is involved in various inflammatory responses and cell death processes. Nevertheless, its function in testicular IRI remains unclear.

METHODS

We employed animal models and the GC-1 cell line to assess tissue damage via HE staining, monitored oxidative stress levels using ROS detection kits, and quantified the key proteins and mRNAs including BRD4, NLRP3, caspase-1, and GSDMD expression through WB alongside qRT-PCR. The secretion of IL-1β/18 was determined by ELISA. Additionally, we utilized the BRD4 inhibitor JQ1 and the NLRP3 inhibitor MCC950, along with siRNA, to knock down BRD4 to investigate their impacts on testicular IRI and pyroptosis.

RESULTS

Our findings indicate that testicular IRI significantly enhances inflammatory responses and pyroptosis. BRD4 expression is significantly upregulated during IRI, and the BRD4 inhibitor JQ1 effectively mitigates testicular IRI by reducing the pyroptosis-related proteins and inflammatory cytokines levels. In cell experiments, knocking down BRD4 improved the survival rate of GC-1 cells, alleviated oxidative stress, lowered the pyroptosis-related protein expression, and reduced the IL-1β/18 secretion. Furthermore, our experiments suggest that BRD4 may promote pyroptosis by modulating the NLRP3 inflammasome pathway.

CONCLUSION

BRD4 exacerbates pyroptosis induced by testicular IRI through the NLRP3 pathway. Inhibition of BRD4 expression or activity may present new therapeutic approaches and targets for testicular IRI prevention and treatment.