Abstract:
Objective To investigate the protective effect of epiberberine against X-ray-induced damage to human keratinocytes (HaCaT) and its potential molecular mechanism.
Methods HaCaT cells were used in this study, and a control group, an epiberberine-alone group, an irradiation model group, and an epiberberine intervention group were established. Cell proliferation activity was assessed by CCK-8 assay. Cell clonogenic capacity was evaluated by colony formation assay. Apoptosis level was determined using the Annexin V-APC/PI double staining method. Intracellular reactive oxygen species (ROS) levels were measured using the DCFH-DA fluorescence probe. γ-H2AX immunofluorescence staining was used to determine DNA double-strand breaks. Transcriptome sequencing and Gene Ontology functional enrichment analysis were performed to explore the molecular mechanisms.
Results Compared with the X-ray irradiation group, epiberberine pretreatment significantly enhanced the survival and clonogenic capacity of HaCaT cells, inhibited radiation-induced apoptosis, effectively reduced intracellular ROS levels, and decreased the number of γ-H2AX foci (P < 0.01). Transcriptomic analysis showed that epiberberine significantly altered multiple biological processes, including cell cycle G1/S phase transition, response to oxidative stress, histone H3-K9 methylation, and mitochondrial respiratory chain complex function.
Conclusion Epiberberine exerts a significant protective effect against X-ray-induced radiation injury in HaCaT cells, and the mechanism may be related to inhibition of apoptosis, alleviation of oxidative stress, reduction of DNA damage, and promotion of DNA repair.
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