Based On OPG And RANKL To Explore The Effect And Mechanism Of Liuwei Dihuang Decoction And Its "tonifying And Reducing" Drugs On The Intervention Of Postmenopausal Osteoporosis Model Rats

Objective:To investigate the effects of Liuwei Dihuangtang and its three reinforcing and reducing herb pairs on the osteoprotegerin(OPG)and the nucleus factor-κB receptor activation factor ligands(RANKL)in kidney and femur of ovariectomized rat models with postmenopausal osteoporosis.It is main to clarify the effect mechanism of postmenopausal osteoporosis,the main compents and the peinciple of compatibility.Methods:1.The 3-4 month SD rats were randomly divided into blank group(sham operation group),model group,Liuwei Dihuangtang group,Corni Fructus-Moutan Cortex group,Rehmanniae Radix Praeparata-Alismatis Rhizoma group,Dioscoreae Rhizoma Poria group,and alendronate group(positive drug group,Except for the blankgroup,all the other rats were ovariectomized to induce the model of postmenopausal osteoporosis.After 16 days,the rats from different groups received intragastric administration for continuous 90 days.The bone density(BMD),blood calcium(Ca)and phosphorus(P),the serum estradiol(E2),bone alkaline phosphatase(ALP),and the tartrate resistant acid phosphatase(TRAP)were determined.2.The weight changes of the rats were observed every week,and the protein expression levels of OPG and RANKL were detected by using immunohistochemical method.3.To separate and determine of morroniside in Liuwei dihuang decoction and its Cornus officinalis-Cortex moutan couple by HPLC.The Hypersile C18 column(4.6mm×250mm,5μm),the mobile phase with methanol-water(24:76),the detective wavelength with 236 nm,the flow rate with 1.0m L/min,the column temperature with30℃ were used in the HPLC method.Results:1.The Liuwei Dihuang Decoction group could decrease the blood TRAP and ALP on model rats,and increase the BMD,the blood Ca and P,and E2.2.As compared with model group,the OPG protein expression in osteoblast and renal tubular epithelial cells was increased,and the protein expression of RANKL in osteoblast and renaltubular epithelial cells was decreased by Liuwei Dihuangtang.The OPG protein expression was increased and the protein expression of RANKL in osteoblast was decreased by Corni Fructus-Moutan Cortex pair.3.The linear ranges of morroniside was 0.480~7.680μg(r = 0.9993),The recovery and relative standard deviation was 102.79%.Conclusion:1.Liuwei Dihuang decoction could improve the level of ALP and TRAP or the content of blood Ca and P were regulated,and it could regulate the metabolism balance of bone by bone formation and bone resorption,and alleviate postmenopausal osteoporosis.The Corni Fructus-Moutan Cortex herb pair is the main effective substance of improving posemenopausal osteoporosis.2.One of the mechanism of Liuwei Dihuangtang for postmenopausal osteoporosis maybe related to regulating OPG and RANKL protein expression in femur by Corni Fructus-Moutan Cortex herb pair.3.The morroniside of Liuwei Dihuangtang is more than Corni Fructus-Moutan Cortex herb pair,it may be one of the reasons that the effect of the Liuwei Dihuangtang on postmenopausal osteoporosis is better than that of Corni Fructus-Moutan Cortex herb pair.