麦角硫因对过氧化氢诱导的成骨细胞损伤的保护作用

目的 探讨麦角硫因(ergothioneine, EGT)对过氧化氢(H₂O₂)诱导的成骨细胞损伤的保护作用及机制。方法 用H₂O₂刺激MC3T3-E1细胞建立氧化应激模型，在加入不同浓度的EGT后，采用CCK8、流式细胞术分别检测细胞活力及凋亡率，利用荧光探针法和WST-8法检测细胞内活性氧(ROS)水平和总SOD酶活性，利用Western blot和RT-qPCR实验分别检测抗氧化及成骨相关蛋白和基因的表达水平，利用碱性磷酸酶染色对细胞成骨能力进行检测。结果 EGT可以剂量依赖地提升氧化损伤环境下MC3T3-E1细胞的存活率，降低H₂O₂引起的ROS生成以及Nrf2、CAT和HO-1的基因和蛋白表达。进一步发现EGT能拮抗H₂O₂对细胞成骨分化的抑制作用。结论 EGT可以抑制H₂O₂引起的氧化应激损伤，并进一步缓解因氧化损伤导致的成骨分化抑制。

The protective effect of ergothioneine on H2O2-induced osteoblast injury

Objective The aim of this study was to investigate the osteoprotective effect of ergothioneine (EGT) on the H₂O₂-induced injury of MC3T3-E1 cells. Methods H₂O₂ was used to establish MC3T3-E1 cells oxidative stress model in vitro, and then co-treated with different concentrations of EGT. CCK8 and flow cytometry were used to detect cell viability and apoptosis. Intracellular reactive oxygen species (ROS) level was detected by fluorescent probe method. The expression levels of antioxidant-related markers were detected by Western blot and RT-qPCR, while total SOD enzyme activity was determined by WST-8. After osteogenic induction, the effects on osteogenic differentiation under oxidative stress condition were detected by Alkaline phosphatase staining, Western blot and RT-qPCR. Results EGT could increase cell viability of MC3T3-E1 cells induced by H₂O₂ in a dose dependent manner, reduced the release of ROS, decreased the protein and mRNA expression levels of Nrf2, CAT and HO-1. In addition, EGT could antagonize the inhibition of H 2O2 on osteogenic differentiation. Conclusion Ergothioneine could inhibit the H₂O₂-induced oxidative damage of MC3T3-E1 cells and promote osteogenesis through anti-oxidative stress.