New Technology Xianlinggubao Preparations On Experimental Osteoporosis Research

Xianlinggubao(XLGB),a Chinese herbal preparation consisting of six herbs including herba epimedii,radix dipsacus,semen psoraleae,radix rehmanniae,radix salviae miltiorrhizae and rhizoma anemarrhenae,has been widely used as herbal treatment for osteoporosis(OP) in clinic.Based on the theory of Traditional Chinese Medicine,this drug posses the effectiveness of activating blood,dissolving stasis,nourishing kidney,and strengthening bone, In this thesis,we systematically investigated the anti-osteoporosis activity of XLGB in OP rats and possible mechanism of action,and compared the efficacy of new artwork with original artwork.1.Anti-osteoporosis activities of XLGB on OVX-induced OP in rats.Methods:(1) Female Wistar rats(3-months-old) were induced by ovariectomy,which were randomized into 8 groups:sham-operated(Sham),ovariectomized(OVX),Premarin(ESG), original artwork XLGB low-dose(X0.5),original artwork XLGB medium-dose(X1),new artwork XLGB low-dose(NX0.5),new artwork XLGB medium-dose(NX1) and new artwork XLGB high-dose(NX2).Daily oral administration of drugs started after OVX for 2 days,lasting for 3 months.Shamand OVX administrated 0.5%CMC,X0.5 and X1 were given original artwork XLGB(0.5 and 1 g·kg^-1·day^-1,respectively),NX0.5,NX1 and NX2 administrated new artwork XLGB(0.5,1,and 2 g·kg^-1·day^-1,respectively).Positive control animals were administrated with premarin 0.03 g·kg^-1·day^-1,wich is used to treat symptoms of menopause.(2) Bone mineral density(BMD) of distal femur and lumbar spine were determined using DEXA.Three-point bending tests were used to investigate the rat femur bone biomechanical parameters.Contents of calcium,phosphorus,magnesium,AKP in serum were measured with automatic biochemical analyzer.E2 and NTX levels in serum were analyzed by means of microparticle enzyme immunoassay method and enzyme-linked immunosorbent assay,respectively.Inductively coupled plasma mass spectrophotometer(ICP-MS) was uised to determine the content of Ca,P,Mg,St,Mn,Zn,Cu in the ashes of rats left femur.Results:(1) The serum E₂ levels and uterus index were significantly lowered in the OVX compared with the Sham.Compared to OVX,the E₂ and uterus index were not changed in new and original artwork XLGB groups,while obviously increased in the ESG group(2) Although ash weight index of distal femur BMD and femur showed significant lower in OVX group,it was high in new and original artwork XLGB groups compared with OVX in rats.The ash weight index in the femur bone was higher in the NX1 than in the X1.(3) Histomorphometric analysis showed a decrease in TBV%but increase in TRS%, TFS%and MAR and OSW and mAR did not change in OVX compared with sham.Both new and original artwork of XLGB can significantly increase the TBV%,reduce the TFS%and MAR,while the new artwork can significantly reduce the TRS%.(4) The maximum load of femur neck was significantly lowered in the OVX than in the Sham.Femur neck fracture load,as well as the maximum load and fracture load in three-point bending did not change significantly in OVX rats.The maximum load of femur neck in XLGB new artwork group was significantly increased compared with OVX rats.(5) Serum AKP and NTX was significantly increased in OVX rats,but reduced in new and original artwork XLGB groups.(6) As compared with sham,serum Ca level was reduced but serum P and Mg content did not change in OVX rats.The bone ash content of Ca,P,Mg,Sr,Mn,Zn and Cu were also decrease& Both new and original artwork XLGB groups can dramatically increase the content of Ca,P,Mg,Sr and Cu.But the new artwork XLGB can significantly increase the content of Zn and Mn,it was higher than original artwork XLGB.(7) The pharmacodynamics of new artwork XLGB Showed a dose-dependent manner.Conclusions:The new artwork XLGB was able to increase bone mass,prevent deterioration of bone microarchitecture,and decrease bone resorption.The new artwork had obvious advantages in decrease bone turnover with original artwork,Meanwhile it had certain ascendancy in improving BMD and collum femoris biomechanics property.The possible mechanisms of XLGB were concerning inhibiting osteoclast activity and decrease bone resorption.2.Anti-osteoporosis activities of XLGB on Tretinoin-indueed OP in rats.Methods:(1) Three-month-old male SD rats were randomized into 8 groups:normal group (Normal),model group(Model),alendronate group(FSM),X0.5,X1,NX0.5,NX1 and NX2. Except Normal group,other animals were treated with retinoic acid(80 mg·kg^-1·day^-1) for 15 days,then drugs were administrated orally once daily for 35 consecutive days.Positive control animals were treated alendronate 0.17 mg·kg^-1·day^-1.Sham and Normal rats were given 0.5%CMC only.(2) Blood,left and right side of tibia,femur and lumbar spine of the rats were gathered. The femur,proximal and distal femur and lumbar spine BMD was determined by DEXA. Proximal tibia microarchitecture was evaluated by bone histomorphometry.Collum femoris mechanical strength was evaluated by compressive testing.Bone turnover biochemical markers serum AKP and StrACP were evaluated with the mothod above mentioned.Results:(1) BMD of femur,lumbar spine,and proximal and distal femur exhibited significant lower in the OP model rats,while femur,proximal and distal femur BMD of new artwork XLGB obvious higher than Model.(2) As compared with Normal,histomorphometric ana!ysis showed a decrease in TBV% but increase in TRS%,TFS%,MAR,mAR,and OSW in Model group.Both new and original artwork of XLGB can significantly increase TBV%,while the TRS%,TFS%,MAR,and mAR were declined in the new artwork-treated model rats.The TRS%was higher in the NX1 than in the X1.(3) The maximum load and energy absorption of femur neck was significantly lowered in the Model than in the Normal.The maximum load of femur neck in XLGB new and original artwork group was significantly increased,while new artwork had a significant decline in energy absorption.(4) Serum AKP and StrACP was significantly increased in Model,but Serum StrACP levels were reduced in new and original artwork XLGB groups,and lowered in the NX1 than in the X1.New artwork XLGB can significantly decrease AKP.Conclusions:the new artwork XLGB was able to increase bone mass and collum femoris biomechanics property,prevent deterioration of bone microarchitecture,and decrease bone resorption.The new artwork had obvious advantages in inhibiting bone turnover comparded with the original artwork XLGB.Meanwhile it had certain ascendancy in improving collum femoris biomechanics property.The possible mechanisms of XLGB may relate to the inhibition of osteoclast activity and bone resorption.