壮骨胶囊调控RAS/RAF/MEK/ERK通路对骨质疏松症大鼠骨代谢的影响OA

Objective:To investigate the effect of Zhuanggu capsule(ZGC)on bone metabolism in rats with osteoporosis(OP)by regulating the rat sarcoma(RAS)/Raf protein kinase(RAF)/mitogen activated protein kinase(MEK)/extracellular regulated protein kinase(ERK)pathway signaling pathway.Methods:A total of 60 rats were randomly divided into a normal group,a model group,a low-dose ZGC(ZGC-L)group(0.5 g/kg),a high-dose ZGC(ZGC-H)group(2.0 g/kg),and a ZGC-H(2.0 g/kg)+ML-099(11.7 mg/kg)group,with 12 rats in each group.After 12 weeks of administration,the bone mineral density(BMD)of the femur was measured in each group.Pathological changes of femoral tissue were detected by HE staining.Femoral cell apoptosis was detected by TUNEL staining.The serum levels of bone-specific alkaline phosphatase(BALP),osteocalcin(OC),core-binding factor-α1(CBF-α1),and procollagen type I N-terminal propeptide(PINP)were measured by enzyme linked immunosorbent assay(ELISA).The expression of proteins related to the RAS/RAF/MEK/ERK signaling pathway was detected by Western blotting.Results:In the normal group,the trabecular bone was arranged neatly and densely with uniform gaps,and the bone tissue structure was intact and regular.In the model group,the trabecular bone gaps were larger,the femoral tissue was sparse and disorderly arranged,and the femoral tissue damage was obvious.In the ZGC-L group and ZGC-H group,the trabecular bone spacing was narrower,the femoral tissue was relatively dense and structurally regular,and the femoral tissue damage was reduced.The degree of femoral tissue damage in the ZGC-H+ML-099 group was higher than that in the ZGC-H group.Compared with the normal group,the model group showed a higher femoral cell apoptosis rate,higher relative expression levels of femoral RAS and RAF proteins,higher p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 ratios(P<0.05),lower femoral BMD,and lower serum levels of BALP,OC,CBF-α1,and PINP(P<0.05).Compared with the model group,the ZGC-L group and ZGC-H group showed a lower femoral cell apoptosis rate,lower relative expression levels of femoral RAS and RAF proteins,lower p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 ratios(P<0.05),higher femoral BMD,and higher serum levels of BALP,OC,CBF-α1,and PINP(P<0.05).Compared with the ZGC-H group,the ZGC-H+ML-099 group showed a higher femoral cell apoptosis rate,higher relative expression levels of femoral RAS and RAF proteins,higher p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 ratios(P<0.05),lower femoral BMD,and lower serum levels of BALP,OC,CBF-α1,and PINP(P<0.05).Conclusion:Zhuanggu capsule may regulate bone metabolism in OP rats by inhibiting the RAS/RAF/MEK/ERK signaling pathway.