Methylene blue alleviates neuronal ferroptosis by down-regulating transferrin after traumatic brain injury

YUE Zheming; WANG Zhibiao; XIE Wenyu; MA Lijun; YONG Jia; XIE Yuan; ZHANG Lei; LI Xia

doi:10.13276/j.issn.2097-1656.2024.08.005

Abstract：Objective To investigate the effects of methylene blue(MB)on iron-mediated cell death in neuronal cells following traumatic brain injury(TBI)and the underlying molecular mechanisms.Methods Hippocampal neuronal HT22 cell line was cultured and randomly divided into blank control group(Con group),injury model group(Model group),and MB treatment group(MB group).In vitro TBI model was used,MB group and Model group were scratched for 6 h,and then cultured for 48 h in normal culture medium supplemented with 2 pomol/L MB.Cell viability was detected using CCK-8 assay,and the levels of iron death markers IL-18,IL-1β,and Fe2+were measured using specific kits.The expression of transferrin(Tf)was examined using fluorescence immunohistochemistry,and the change of Tf expression level was measured by Western blotting.Results Compared with Con group,HT22 cell viability was significantly decreased,the levels of IL-18,IL-1β,and Fe2+were significantly increased,and Tf protein expression was increased in Model group,with statistical significance(P＜0.05).In comparison with Model group,MB group exhibited increased cell viability,decreased levels of IL-18,IL-1β,and Fe2+,and decreased expression of Tf protein,as well as a significant reduction in Tf staining positivity in cytoplasm and cell membrane,with statistical significance(P＜0.05).Conclusion MB interferes with iron death-related molecules by regulating Tf expression,alleviates TBI-induced hippocampal neuronal damage,inhibits neuronal ferroptosis,and plays an important role in the treatment of TBI.