To Explore The Mechanism Of Deer Antler Glue In The Treatment Of Osteoporosis Based On OPG-RANKL-RANK Pathway

Objective: This study investigates the effect of deer horn gum on bone resorption in mice with osteoporosis(OP),confirms the efficacy of deer horn gum from the perspective of osteoclast(OC),and investigates the mechanism of action of deer horn gum on OP from in vivo and in vitro experiments.Methods: In vivo:C57BL/6female mice aged 8 weeks were randomized into 4 groups:(1)SHAM(Sham operation)+normal saline(NS),(2)SHAM+Deer antler glue(DAG),(3)OVX+PS and(4)OVX+DHG(n=6 in each group).After 4 weeks continuous administration,changes of femoral bone mass were determined by HE staining,TRAP staining and micro-CT.The concentrations of OPG and RANKL in serum were determined by ELISA.The protein expressions of OPG and RANKL were determined by Western Blot.In vitro: 8-week-old female SD rats were randomly divided into blank serum group(Bs,n=6)and deer antler glue drug-containing serum group(Ds,n=6).After 7days of deer antler glue administration,Fetal calf serum,blank serum and deer antler glue drug-containing serum were prepared for culturing osteoclasts.Tartrate-resistant acid phosphatase 5b(TRACP-5b)was used to determine osteoclast viability for both groups.Results:(1)After 4 weeks of Chinese medicine intervention,HE staining and micro-CT of mice femur showed that the OVX+PS group had less and narrower trabeculae with greater spacing than that in SHAM+PS group,indicating a successful establishment of osteoporotic animal model.Compared with OVX+PS group,the trabeculae in OVX+DHG group was thicker and wider,and the BV/TV and Tb.N were significantly increased(P<0.01).(2)Compared with OVX+PS group,OVX+DHG group was observed serum RANKL concentration decreased and OPG concentration increased,RANKL protein expression decreased and OPG protein expression increased(P<0.05).(3)Compared with Bs group,the number and area of TRAP-stained osteoclasts were decreased,and the activity of osteoclasts was inhibited in Ds group(P<0.05).Conclusion: Results show that deer antler glue effectively inhibited the activity of osteoclasts and osteoclastogenesis,while promoting the increase of bone mass,the thickness of trabeculae,as well as the improvement of trabecular bone structure.The clinical application of deer antler glue may be one of the important methods for treating osteoporosis.