gammaH2AX foci analysis for monitoring DNA double-strand break repair: strengths, limitations and optimization.

BACKGROUND

Cisplatin is widely utilized in the treatment of solid malignant tumors due to its potent anticancer effects through the inhibition of cell division. However, its clinical use is often limited by significant adverse effects, particularly nephrotoxicity. Recent research has focused on natural products as potential mitigators of cisplatin-induced kidney toxicity. Allium hookeri (A. hookeri), a traditional food and herbal medicine in Southeast Asia, is known for its antioxidant and anti-inflammatory properties. However, its protective effects against nephrotoxicity remain unclear.

OBJECTIVE

This study aimed to investigate the protective effects of A. hookeri against cisplatin-induced nephrotoxicity in human embryonic kidney (HEK)-293 cells.

METHODS

HEK-293 cells were treated with cisplatin (50 μM) with or without A. hookeri water extract (AHWE) and ethanol extract (AHEE) for 24 h. Cell viability was assessed using MTT assays, and nuclear morphology was examined through Hoechst 33342 staining. Intracellular reactive oxygen species (ROS) production was quantified using ROS detection assays, and nitric oxide (NO) production was measured through Griess reaction assays. Protein and mRNA expression levels were analyzed using western blotting and quantitative polymerase chain reaction (qPCR) techniques.

RESULTS

Cisplatin treatment (50 μM) significantly increased ROS production compared to untreated cells within 24 h. Both AHWE and AHEE treatments markedly attenuated ROS generation. Additionally, AHWE and AHEE significantly inhibited NO production and downregulated the expression of inflammation-related genes. The treatments also suppressed mitogen-activated protein kinase (MAPK) protein expression. Pretreatment with AHWE and AHEE decreased the Bax/Bcl-2 expression ratio, demonstrating a dose-dependent inhibition of apoptotic features.

CONCLUSION

The findings suggest that A. hookeri exerts protective effects against cisplatin-induced kidney damage by modulating MAPK signaling, thereby reducing inflammation and apoptosis in HEK-293 cells. A. hookeri represents a promising therapeutic candidate for the prevention and treatment of nephrotoxicity.