Effect On Bushen Jianpi Huoxue Recipe On TGF-β1/Smads Signaling Pathway In Ovariectomized Rat Bone Tissue

Objective1.To observe the expression and effect of Bushen Jianpi Huoxue Recipe on transforming growth factor-β1(TGF-β1),Smads-related genes and proteins in ovariectomized rats.2.To explore the mechanism of action of Bushen Jianpi Huoxue Recipe on TGF-β1/Smads signaling pathway and the prevention and treatment of OP.Methods1.Modeling:72 female Sprague-Dawley rats aged 6 months were randomly divided into sham operation group(n=24).The operation group(n=48)was modeled by bilateral ovarian resection.After 3 months,the two groups were randomly selected 12 samples were tested for bone density to verify successful modeling.2.Grouping and drug intervention:The remaining 36 surgical rats were randomly divided into model group(OVX),Bushen Jianpi Huoxue Recipe group(BSJPHX,administered dose 2.979 g·kg-1),alendronate Sodium-dimensional D3 tablets(Ⅱ)group(ALN,dose 1.02 mg·kg-1).Sham operation group and OVX group were given equal volume of normal saline.3.Indicator detection:rats were sacrificed after 3 months of drug intervention.Bone mineral density was measured by dual-energy X-ray method.Histopathological changes were observed by HE staining.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)detection of TGF-β1,Smad2,Smad3,Smad4,Smad7,osteoprotegerin(OPG),Runt-related transcription factor 2(Runx2)mRNA expression,Western blot analysis of TGF-β1,Smad2,Smad3,Smad4,Smad7,OPG,Runx2 protein expression.Result1.Model validation and bone mineral density test:After 3 months of modeling,compared with the normal group,the bone mineral density of the OVX group was significantly reduced,indicating that the rat model of osteoporosis was successful.After 3 months of drug intervention,bone mineral density increased in the BSJPHX group and the ALN group compared with the OVX group(P<0.05).2.Effect of Bushen Jianpi Huoxue Method on Femoral Tissue Morphology in RatsThe results of HE staining showed that the sham operation group was thick and tidy,the arrangement was relatively neat,the morphological structure was relatively complete,and the structure of the bone marrow cavity was normal.In the OVX group,the trabecular bone was broken,disordered,.and the shape was incomplete,and the cavity of the bone marrow was visible.After administration,the bone marrow cavity of the BSJPHX group and the ALN group became smaller,the number of trabecular bones was larger,the arrangement was more neat,and the structure was tight.3.Effects of Bushen Jianpi Huoxue Method on TGF-p 1,Smad2,Smad3,Smad4,Smad7,OPG,Runx2 mRNA and Protein in Rat FemurCompared with the OVX group,the BSJPHX group and the ALN group significantly increased TGF-β1,Smad2,Smad3,Smad4,OPG,Runx2 mRNA and protein expression levels(P<0.05);decreased Smad7 mRNA and protein expression levels(P<0 05).Conclusion:1.Bushen Jianpi Huoxue Recipe can increase the bone density of femur in ovariectomized rats,promote bone formation,increase the expression of TGF-β1,Smad2,Smad3,Smad4,OPG,Runx2 mRNA and protein in bone tissue,and decrease the expression of Smad7 mRNA and protein.2.Bushen Jianpi Huoxue Recipe may play a preventive and therapeutic role in osteoporosis by regulating the transduction of TGF-β1/Smads signaling pathway.