Action Mechanism of Osteonecrosis Rehabilitation Pill in Treatment of Hormonal Femoral Head Necrosis
Objective:Based on the relationship between phosphatidyl inositol 3-kinase(PI3K)/protein kinase B(PKB,also known as Akt)/hypoxia inducible factor-1α(HIF-1α)signaling pathway and glycogen synthase kinase-3 beta(GSK-3β)signaling path-way to explore the mechanism of action of Osteonecrosis Rehabilitation Pill in treatment of steroid-induced osteonecrosis of the femoral head(SONFH).Methods:Twelve rats were randomly selected from 65 male rats as a blank group,and the rest of the rats were injected with lipopolysaccharide(20 μg·kg-1)in combination with methylprednisolone(40 mg·kg-1)to establish the SONFH model.The rats successfully modeled were randomly divided into the model group,Xianling Gubao Capsule group(0.315 g·kg-1),Osteoradione-crosis Rehabilitation Pill high-dose group(27.30 g·kg-1),and the medium-dose group(13.65 g·kg-1),with 12 rats in each group,and were gavaged consecutively for 6 weeks.Micro-CT was used to observe the structure of the femoral head of rats,HE staining was used to observe the pathologic morphology of the femoral head,and ELISA was used to detect the content of serum HIF-1α.Bone marrow mesenchymal stemcells(BMSCs)were grouped in the same way as animal experiments,and the cell proliferation rate was de-tected by CCK-8 method,and the protein expression levels of GSK-3β,HIF-1α and Akt were detected by Western blot method.Re-sults:Micro-CT results showed that compared with the blank group,the bone trabeculae of the femoral head of rats in the model group were short and sparse,and the tissue microstructure was obviously damaged.Compared with that of the model group,the structure of tra-beculae in the drug group was improved,and the arrangement tended to be regular.Compared with that of the blank group,the bone vol-ume fraction,bone mineral density,bone trabecular thickness and the number of bone trabeculae in the model group were reduced(P＜0.01).Compared with that of the model group,bone volume fraction,bone mineral density and number of trabeculae increased in the high dose group(P＜0.05).HE staining results suggested that the cartilage layer of the femoral head of the rats in the blank group was thicker,the trabeculae were neatly arranged,and a large amount of active proliferative bone marrow tissue could be seen in the medullary cavity.In the model group,the cartilage layer was thinner,the bone trabeculae were sparse and disorganized,and many necrotic tissue fragments were seen in the medullary cavity,accompanied by many vacant bone sockets and osteonecrosis.Compared with that of the model group,the rats in the drug group showed improved disorganization and fracture of bone trabeculae,and fewer vacant bone sockets.Compared with that of the blank group,the serum HIF-1 α content of rats in the model group was decreased;compared with that of the model group,the serum HIF-1α content of rats in the drug group was increased(P＜0.01).With CCK-8 assay,it was suggested that the proliferation rate of BMSCs in the model group was decreased compared with the blank group(P＜0.05).Compared with the model group,the proliferation rate of BMSCs was increased in Xianling Gubao Capsules group and Osteonecrosis Rehabilitation Pill high-dose group(P＜0.01).Western blot results indicated that the expression levels of GSK-3β,HIF-1α and Akt of BMSCs in the model group were decreased compared with the blank group(P＜0.01).Compared with that of the model group,the expression levels of GSK-3β,HIF-1α,and Akt in BMSCs in the drug group were increased(P＜0.01).Conclusion:Osteonecrosis Rehabilitation pill may exert a therapeutic effect on SONFH by regulating PI3K/Akt/HIF-1α signaling pathway and GSK-3 β signaling pathway.
hormonal osteonecrosis of the femoral headOsteonecrosis Rehabilitation PillPI3K/Akt/HIF-1α signaling pathwayGSK-3 β signaling pathwayratbone marrow mesenchymal stem cell
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