| Objective:By studying the effect of zhuangguzhitong capsuleon the expression of bone formation related proteins in castrated rats and the effect of drug-containing serum on the expression of osteoblast related proteins,the mechanism of zhuangguzhitong capsuleon in regulating bone formation was discussed.Methods:In this study,the pathological model of postmenopausal osteoporosis was prepared by bilateral ovariectomy in rats,which was randomly divided into blank group,model group,gusongbao group and zhuangguzhitong capsule groups by body mass,including 10 groups with high dose,medium dose and low dose.After 13 weeks of intervention with zhuangguzhitong capsuleon of various concentrations,the blank group and the model group were given equal volume distilled water.After the intervention,blood samples from abdominal aorta were collected to prepare drug-containing serum for later use.Rat femur was stained with HE for bone histopathological and morphological detection.Rat femur of immunohistochemical method to detect bone-building genes specific transcription factor Runt related gene 2(Runt related transcription factor 2,Runx2),osteogenesis related transcription factor antibody(osterix,Osx),Dickkopf associated protein 1(Dickkopf related protein 1,DKK1)expression;A method was established for the extraction and culture of primary osteoblasts in neonatal SD mice,which were cultured until the third generation,and then added with 10%serum of the corresponding concentration,which was stained with silver and alkaline phosphatase(ALP)to identify osteoblasts,and the expressions of Osx and osteopontin(OPN)in osteoblasts were detected by immunohistochemistry.Results:1.In vivo experiment,compared with the blank group,the trabecular area of the model group decreased,and the ratio of trabeculae to marrow decreased significantly.Immunohistochemical results showed that the expressions of Runx2 and Osx proteins in femoral tissue decreased,while the expression of DKK1 protein increased.Compared with the model group,trabecular area and girder pulp ratio were improved in the gusongbao group and zhuangguzhitong capsuleon high-dose,medium-dose and low-dose groups(P<0.05).The expressions of Runx2 and Osx protein in femoral tissue increased(P<0.05),while the expression of DKK1 protein decreased(P< 0.05).Moreover,the effect of zhuangguzhitong capsuleon at high dose was similar to that of the positive drug gusongbao without statistical difference(P>0.05).2.In vitro experiments,compared with the blank group,the expression rates of Osx and OPN proteins in the model group were significantly reduced(P<0.05).Compared with the model group,Osx and OPN protein expressions were increased in the gusongbao group,zhuangguzhitong capsuleon high-dose,medium-dose and low-dose groups(P<0.05).Conclusion:1.The trabecular area and girder pulp ratio of femur of castrated rats were significantly decreased,the expressions of Runx2 and Osx proteins in femur tissue were significantly decreased,and the expression of DKK1 protein was significantly increased,indicating that the model building was successful and the bone formation effect was weakened.2.zhuangguzhitong capsuleon can up-regulate the expression of Runx2 and Osx proteins in femoral tissue and down-regulate the expression of DKK1 protein,which may be one of the mechanisms of its anti-postmenopausal osteoporosis.3.zhuangguzhitong capsuleon drug-containing serum can promote the secretion of Osx and OPN proteins in osteoblasts,thereby affecting the differentiation and proliferation of osteoblasts and playing an anti-menopausal role in osteoporosis. |
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