Effects Of Puerarin,Tanshinone ?A Combined With VitD₃ On Osteoporosis Induced By Retinoic Acid In Rats

Postmenopausal osteoporosis is a metabolic disease,due to deterioration of ovarian function in postmenopausal women and a sharp decline in estrogen levels in the body,resulting in increased osteoclast activity and increased bone turnover rate,indicating decreased bone mass,decreased bone density,decreased bone fragility.In clinical treatment,estradiol is often used to treat postmenopausal osteoporosis and reproductive system diseases.Estrogen can directly increase estrogen levels in the body,stimulate osteoblast activity,correct bone metabolism abnormalities,and relieve atrophy of the reproductive system.However,long-term use of chemically synthesized estrogens can easily lead to cervical cancer,ovarian cancer,endometrial cancer,and cardiovascular thrombosis.Therefore,the use of phytoestrogens to treat osteoporosis is beneficial,but studies have shown that the efficacy of simple use of phytoestrogens is not ideal.In this study,two phytoestrogens,puerarin and tanshinone IIA,were selected,which were administered to rats in combination with VitD₃ for osteoporosis induced by retinoic acid,respectively.On the one hand,to observe whether puerarin and tanshinone IIA can exert estrogen-like effects,prevent bone loss,promote bone formation,and reduce bone fragmentation.On the other hand,it was observed whether the efficacy of puerarin,tanshinone IIA and VitD₃ were superior to administrated alone.Experimental animals were divided into model group,normal group,puerarin group?PUE group?,tanshinone IIA group?TSIIA?,tanshinone in combination with VitD₃ group?TSIIA+VitD₃?,puerarin in combination with VitD₃?PUE+VitD₃?and estradiol?E₂?group.After 70 days of administration,blood biochemical indicators?E₂,OPG,RANKL,ALP?,body weight,organ index,bone histomorphometry,osteoblasts and osteoclast number,OPG,RANKL,RUNX2protein and mRNA expression were deceted.At the same time,the effects of puerarin and tanshinone II A on the uterus and vaginal tissue in osteoporotic rats were observed by histomorphometric methods.This study will provide a theoretical basis for increasing BMD health foods and drug development,and reproductive system repairment in the patients of postmenopausal osteoporosis.The main results were as follows:?1?Models of rat osteoporosis could be established successfully by intragastric administration of retinoid 80 mg/?kg·d?for 21 days.Compared with the normal group,the model group's body weight and organ index were significantly reduced.Serum alkaline phosphatase was significantly elevated.The bone microstructure was destroyed.Bone trabecula became thinner,and fractured.Trabecular spacing was increased,and the bone marrow fat cells were increased.?2?ELISA analysis of serum OPG,RANKL,ALP and E₂ in each group showed that tanshinone IIA combined with vitamin D₃ could significantly increase serum OPG and E₂ levels in osteoporosis rats,and significantly reduce serum RANKL and ALP levels.The ratio of OPG/RANKL was significantly higher than tanshinone group,puerarin group and puerarin combined with vitamin D₃ group.?3?Histomorphometric analysis of femur metaphysis showed that the trabecular bone in the model group was thinner,and the spacing was larger.The trabecular bone was fractured.And the area of the trabecular bone was reduced.Puerarin and tanshinone IIA combined with vitamin D₃ could effectively improve the bone quality,and bone structure,increase trabecular thickness,and percentage of trabecular area,decrease trabecular spacing.Puerarin and tanshinone IIA combined use of VitD₃ were superior to its use alone.The bone protect effects of TSIIA+VitD₃ was better than PUE+VitD₃.?4?Osteoblasts and osteoclasts were stained with an enzyme.The results of immunohistochemistry and density measurements of OPG,RUNX2 and RANKL,in bone tissue showed that puerarin,puerarin combined with vitamin D₃,tanshinone IIA,and tanshinone IIA combined with vitamin D₃ can enhance the number of osteoblasts and the expression of OPG,RUNX2,and can inhibite the number of osteoclasts and RANKL expression in bone tissues.?5?The detection results of OPG mRNA,RANKL mRNA and VDR mRNA in bone tissue showed that the up-regulation of OPG mRNA and VDR mRNA and the suppression of RANKL mRNA by tanshinone IIA combined with VitD₃ were superior to other groups.The up-regulation of osteogenesis and osteogenic gene expression in tanshinone IIA combined with VitD3 group were superior to puerarin and puerarin combined with VitD₃ group.?6?Uterine,vaginal histopathology and morphometric analysis showed that puerarin and tanshinone IIA could significantly increase the wet weight of reproductive organs in rats with osteoporosis,increase serum E₂ levels,and improve pathology of the uterus and vagina of rats with osteoporosis.puerarin and tanshinone IIA could exert estrogen-like effects and have a repairing effect on the reproductive system of osteoporosis rats.