葛根素通过NF-кB信号通路调节破骨细胞的分化

Puerarin inhibits osteoclast differentiation through NF-кB signal pathway

目的探索葛根素抑制破骨细胞分化的实验研究。方法 RAW264.7细胞经sRANKL+M-CSF诱导7 d后向破骨细胞分化;WST-1检测不同浓度的葛根素对破骨细胞增殖活性的影响;TRAP染色检测葛根素对破骨细胞分化的影响;Osteoassay surface Mltiple well plate检测破骨细胞吸收活性;免疫荧光检测葛根素对NF-кB通路核心蛋白p65细胞核内转移的影响;Western blot检测葛根素对p65表达的影响。结果葛根素(10-8mol/L)作用3 h后破骨细胞的增殖活性明显抑制,葛根素抑制破骨细胞的分化和吸收活性,免疫荧光检测显示葛根素抑制p65的细胞核内转位及p65的磷酸化。结论葛根素通过NF-кB信号通路抑制破骨细胞的分化。

Objective To explore the mechanism of Puerarin inhibiting osteoclast differentiation.Methods The RAW264.7 cells were induced by sRANKL+M-CSF for 7 days to differentiate into osteoclasts.MTS detected the effects of Puerarin on the proliferation of osteoclasts with different concentrations,and the effect of Puerarin on osteoclast differentiation was detected by TRAP staining.The absorptive activity of osteoclasts was detected by osteoassay surface multiple well plate,and the effects of Puerarin on the intracellular transfer and expression of the core protein p65 of NF-кB signaling pathway was detected by immunofluorescence and Western blot,respectively.Results MTS showed that Puerarin(10-8 mol/L,3 h)significantly inhibited the proliferation of osteoclasts.Puerarin obviously inhibited the differentiation and the absorptive activity of osteoclasts,inhibited the nuclear transposition of the core protein p65,and inhibited the p65 phosphorylation.Conclusion Puerarin inhibits osteoclast differentiation through NF-кB pathway.