The Mechanism Of Bushen Tongluo Formula In Improving Postmenopausal Osteoporosis Based On Chemerin

Objective:To evaluate the effects of Bushen recipe,Tongluo recipe and Bushen Tongluo Recipe on bone remodeling in postmenopausal osteoporosis model and bone formation and bone resorption model in vitro,and to explore whether Bushen Tongluo recipe regulates bone remodeling through Chemerin.Methods:(1)84 SD rats were randomly divided into 7 groups(n=12)and 56 Chemerin knockout(Rarres2-/-)mice were randomly divided into 7 groups(n=8),Sham operation group,model group,alendronate sodium group,sodium fluoride group,Bushen group,Tongluo group and Bushen Tongluo group were set up.They were given saline,alendronate sodium suspension,sodium fluoride solution,Bushen recipe,Tongluo recipe and Bushen Tongluo Recipe at once a day for 8 weeks.(2)MC3T3-E1 osteogenic model and RAW264.7 osteoclastic differentiation model were established.Control group,Bushen group,Tongluo group and Bushen Tongluo group were set up respectively.After 24 hours of intervention,the expression of related genes and proteins were detected.(3)MC3T3-E1 osteogenic model and RAW264.7 osteoclastic differentiation model were established after Chemerin silencing.Blank control group,BS group,TL group and BSTL group were set up respectively.Each group was given corresponding Chinese medicine,and the expression of related genes and proteins were detected after 24 hours of intervention.(4)MC3T3-E1 osteogenic model and RAW264.7 osteoclastic differentiation model were established.The control group,Chemerin+BS group,Chemerin+TL group and Chemerin+BSTL group were set up respectively.The corresponding drugs were given to each group.After 24 hours of intervention,the expression of related genes and proteins were detected.Results:(1)Compared with the control group,the weight of rats and Chemerin-/-mice in the model group increased significantly;compared with the model group,the weight of rats and mice in the BSTL group did not change significantly.(2)Compared with the control group,the BMD of the model group and Chemerin-/-mice were significantly decreased,and the levels of serum inflammatory factors were significantly increased;compared with the model group,the BMD of the BSTL group was significantly increased,the BMD of the bstl group Chemerin-/-mice was significantly increased,the expression of serum TNF-α in the BSTL group was significantly decreased,and the levels of serum IL-1,IL-6 and TNF-α in the BSTL group Chemerin-/-mice were significantly increased.(3)Compared with the control group,the osteogenic related proteins and genes in the model group and Chemerin-/-mice were significantly decreased;compared with the model group,the protein RANKL/OPG ratio in the BSTL group was significantly decreased,and the expression of β-catenin mRNA and protein was significantly increased;the osteogenic related genes Runx2,OPG and β-catenin in the BSTL group were significantly increased.The mRNA expressions of TRAP,CTSK,RANKL and RANK were significantly decreased.(4)In normal MC3T3-E1 cells,compared with the control group,the expressions of OPG,β-Catenin and the mRNA expressions of β-catenin,Runx2,OPG and BALP were significantly increased in BSTL group;in Chemerin silenced MC3T3-E1 cells,compared with the control group,the protein and gene expressions of β-catenin and OPG were significantly increased,and the mRNA expressions of BALP and Runx2 were significantly increased in BSTL group The expression of OPG,β-catenin,Runx2 and BALP mRNA in MC3T3-E1 cells treated with Chemerin was significantly higher than that in the control group.(5)In normal RAW264.7 cells,compared with the control group,the expression of RANK protein and mRNA,and the expression of TRAP,CTSK and Chemerin mRNA were significantly decreased in BSTL group;in Chemerin silenced RAW264.7 cells,compared with the control group,the expression of RANK protein and mRNA,and the expression of RANK,TRAP and CTSK mRNA were significantly decreased in BSTL group In Chemerin treated RAW264.7 cells,compared with the control group,the mRNA levels of RANK,TRAP and CTSK in BSTL group were significantly decreased.Conclusions:(1)Bushen Tongluo recipe can promote the expression of osteoblast related genes and proteins through Wnt/β-catenin pathway,and promote bone formation;Bushen Tongluo recipe can inhibit the differentiation and maturation of osteoclasts through RANKL/RANK/OPG pathway,so as to inhibit bone resorption and improve bone remodeling.(2)In Chemerin knockout mice,Bushen Tongluo recipe can promote bone formation and inhibit bone resorption through Wnt/β-catenin and RANKL/RANK/OPG pathways,respectively.(3)In vitro experiments showed that Bushen Tongluo Decoction could promote the expression of bone formation related genes and inhibit the expression of bone resorption related genes by inhibiting Chemerin.After Chemerin intervention,osteogenesis related genes decreased and osteoclast related genes increased.Bushen Tongluo Decoction could reverse the effect,which increased osteogenesis related genes and decreased osteoclast related genes.