雷公藤甲素通过铜死亡信号通路抑制HaCaT细胞的增殖OA

Objective To investigate the effect of tripolide(TP)on the proliferation of HaCaT cells,and to identify a novel mechanism underlying the anti-psoriatic activity of TP.Methods HaCaT cells were treated with 20 ng·mL-1 tumor necrosis factor-α(TNF-α)for 24 h before subsequent experiments.The effect of TP on HaCaT cell proliferation was assessed using CCK-8 and EdU assays.Reverse transcription quantitative real-time PCR,cellular copper colorimetry,and an ATP assay kit were used to detect the mRNA levels of interleukin(IL)-1β,IL-6,IL-23A and TNF-α,the level of copper ions,and the ATP content in HaCaT cells,respectively.Western blotting,reactive oxygen species(ROS)detection,and mitochondrial membrane potential assay were employed to evaluate the effects of TP alone or in combination with tetrathiomolybdate(TTM)on the expression of cuproptosis-related proteins,ROS production,and mitochondrial membrane potential in HaCaT cells.Results TP significantly inhibited HaCaT cell proliferation(P<0.05),markedly suppressed the mRNA levels of IL-1β,IL-6,IL-23A,and TNF-α(P<0.05),increased copper ion levels(P<0.01),and decreased ATP content(P<0.05).Moreover,TP significantly downregulated the expression of ferredoxin 1,succinate dehydrogenase complex iron-sulfur subunit B,DNA polymerase delta 1,and dihydrolipoamide S-acetyltransferase(P<0.05),promoted ROS generation,and reduced mitochondrial membrane potential(P<0.05).TTM partially reversed the TP-induced abnormalities in the expression of cuproptosis-related proteins.Conclusion TP could inhibit the excessive proliferation of TNF-α-induced HaCaT cells via cuproptosis-related signaling pathways,thereby exerting its anti-psoriatic effect.