Study On The Mechanism Of Mongolian Medicine Echinops In Preventing And Treating Postmenopausal Osteoporosis Based On ER/AKT/ERK Pathway In Bone Marrow Mesenchymal Stem Cells

OBJECTIVE The postmenopausal osteoporosis(PMOP)model was established and used to observe the effects of Mongolian Medicine echinops on the expression of ERa and ERβ in mRNA and protein levels,and expression of p-AKT,p-ERX in bone marrow mesenchymal stem cells derived from ovariectomized osteoporosis rats,and on the serum levels of E2,alkaline phosphatase(ALP)and testosterone,which could provide experimental basis for using Mongolian medicine to treat postmenopausal osteoporosis.METHODS Eighty-four female Wistar rats aged 4 months were randomly divided into two groups:sham operation group(Sham),model group(OVX),diethylstilbestrol group,echinops high,medium and low dose groups.In each group of 10 rats.The Sham group only removed a small amount of adipose tissue,and the other groups removed the bilateral ovaries.After successful modeled,the rats were given the corresponding drugs for 90 days after normal feeding,and the rats were sacrificed after 90 days of continuous gavage.Take blood from heart,the serum E2,alkaline phosphatase(ALP)and testosterone levels were measured by ELISA.qRT-PCR was used to detect the expression of ERaand ERβmRNA in each group.Hematoxylin and eosin(H&E)and toluidine blue O staining were performed on the left medial metaphysis.Bone marrow-derived bone marrow srem cells(BMSC)were isolated and treated with serum contained echinops.The cell vialality of BMSCs was detected by CCK8,and observed the colony formation of BMSCs effec.ted by echinops.The expression of ERpand ERβmRNA in BMSCs treated with echinops serum was detected by qRT-PCR.Western blot was used to detect the expression of ERa,ERβ,AKT,p-AKT,ERK and p-ERK in BV MSCs.Bone mineral density(BMD)was measured by micro-CT to detect the left m edial proximal metaphyseal metatarsal humerus.Results Echinops can significantly improve the general condition of ovariectomized osteoporosis rats;echinops can significantly increase E2 levels and decrease ALP and testosterone levels in ovariectomized rats.The expression of ERaand ERβwas significantly increased.Compared with the Sham group,the HE and toluidine blue O staining showed typical osteoporosis and widened intervertebral space.Compared with the OVX group,the echinops increased the trabecular bone connection,improve the thickness of trabecular bone.Serum from the high dose group of echinops can significantly increase cell viability.Echinops increased the expression of ERa,ERβ,p-AKT and P-ERK in BMSCs in a dose-dependent manner.The BMD of the high-dose group echinops increased significantly.Conclusion Echinops increased BMD and E2,but decreased ALP and testosterone in a dose-dependent manner.It can induce the expression of ERaand ERβmRNA and enhance the cell viability and colony forming ability of BMSC.It can affect the expression of ERaand ERβmRNA and ERa,ERβ,p-AKT and P-ERK proteins expression in BMSC.Echinops can inhibit OVX-induced bone loss caused by estrogen deficiency.It indicates that the echinops may regulate the bone formation,regulated the coupling of bone resorption and bone formation,reduced the bone turnover rate,and achieved the purpose of preventing PMOP.