Exploring the Regulatory Mechanism of Jiangu Granules Containing Serum on Bone Formation of Osteoblasts Under Oxidative Stress Status Based on the Nrf2/HO-1 Pathway
Objective:The regulatory mechanism of serum containing Jiangu Granules on bone formation of osteoblasts under oxidative stress was discussed based on Nrf2/HO-1 pathway.Methods:A total of forty SPF male SD rats aged 6 weeks were randomly divided into Jiangu Granules group and normal saline group,with 20 rats in each group.Jiangu Granules group was given 2 mL of Jiangu Granules extract diluent containing 7.8 g/(kg·d)of crude drug,while saline group was given 2 mL of saline,by gavage once a day respectively.After 7 days of gavage,the abdominal aorta blood of rats was taken to prepare Jiangu Granules drug-containing serum and saline serum.The Nrf2 knock-down stable cells and Nrf2 negative control cells constructed by UMR-106 cells were di-vided into Nrf2-KD+JG group,Nrf2-KD+NS group,NC+JG group and NC+NS group respectively.The Nrf2-KD+JG and NC+JG groups were cultured with high glucose DMEM containing 10%serum of Jiangu Granules for 12 h,while the Nrf2-KD+NS and NC+ NS groups were cultured with high glucose DMEM containing 10%saline serum for 12 h,and then were intervened with 10 μmol/L hydrogen peroxide for 12 h.Dihydroethidium(DHE)fluorescent probe was used to detect the content of superoxide anion in four groups,alizarin red staining experiment was used to observe the number of mineralized nodules in four groups,and Western blot was used to detect the relative expression of Nrf2,nuclear Nrf2,HO-1 and RUNX2 proteins in four groups.Results:Compared with the NC+JG group,the content of superoxide anion in the Nrf2-KD+JG group increased(P<0.05),the number of mineralized nodules decreased under microscope,and the relative expressions of Nrf2,nuclear Nrf2,HO-1 and RUNX2 proteins decreased significantly(P<0.05).Compared with the NC+NS group,the content of oxide anions in the Nrf2-KD+NS group increased,the number of mineralized nodules decreased under microscope,and the relative expression of four index proteins decreased significantly(P<0.05).The content of superoxide anion in the NC+JG group decreased significantly(P<0.05),the number of mineralized nodules increased under microscope,and the relative expression of four index proteins increased significantly(P<0.05).Conclusion:Jiangu Granules can activate the Nrf2/HO-1 signaling pathway in osteoblasts,reduce the level of intracellular oxidative stress,enhance the mineraliza-tion ability of cells and promote the bone formation of osteoblasts.
万方数据
维普
