温肾壮骨方对来曲唑诱发骨损伤大鼠的骨保护作用

Protective effects of Wenshen Zhuanggu Formula on letrozole-induced bone injury in rats

目的:温肾壮骨方对来曲唑诱发骨损伤大鼠的骨保护作用。方法:30只SPF级雌性SD大鼠随机分为正常组、模型组和给药组,每组10只。除正常组外,其余组灌胃来曲唑,给药组每日灌胃温肾壮骨方6.5 g/kg,连续24周。动物麻醉后,称重、解剖,采集血液。测定体质量、脏器重量及系数,以及血清氧化应激水平、甲状腺功能、第二信使cAMP与cGMP、NF-κB受体活化因子配体(RANKL)与骨钙素含量。左侧股骨进行抗酒石酸磷酸酶染色和骨钙素免疫组化,右侧股骨检测骨密度。结果:给药组第18周后可显著降低模型组大鼠体质量(P<0.05),剖后发现给药组可增加子宫脏器系数(P<0.05),提高血清游离甲状腺素、甲状腺素、cAMP水平以及cAMP/cGMP比值(P<0.05,P<0.01),降低血清丙二醛、RANKL以及RANKL/骨钙素比值(P<0.01,P<0.05),减少破骨细胞分化(P<0.05),增加成骨细胞分化和骨密度(P<0.05)。结论:温肾壮骨方可通过提高第二信使cAMP与cGMP比值以及甲状腺功能,改善卵巢退化,最终降低大鼠骨质破坏进程。

Objective:To explore the effects of Wenshen Zhuanggu Formula on bone injury in rats with bone loss induced by letrozole.Methods:Thirty SPF grade female SD rats were randomly divided into three groups:normal group,model group and treatment group,with 10 rats in each group.Other groups were given letrozole by intragastric administration except the normal group.The treatment group was given intragastric administration of 6.5 g/kg every day for 24 weeks.After the mice were weighed and anesthetized,blood was collected.The levels of oxidative stress and thyroid function in serum were determined by biochemical analyzer,and the serum levels of cyclic adenosine phosphate(cAMP),cyclic guanosine phosphate(cGMP),NF-κB receptor activating factor ligand(RANKL)and osteocalcin were determined by ELISA.The left femur was fixed by formalin and stained by TRAP staining and OCN immunohistochemical staining.Bone density of right femur was detected by a small animal PET-CT.Results:After 18 weeks,compared with the model group,the treatment group significantly reduced animal body weight(P<0.05).After dissection,the treatment group increased the ratio of uterus to viscera(P<0.05),increased serum FT4,T4 and cAMP levels,as well as cAMP/cGMP ratio(P<0.05,P<0.01),decreased serum malondialdehyde,RANKL and RANKL/osteocalcin ratio(P<0.01,P<0.05),significantly decreased osteoclast differentiation(P<0.05)and increased osteoblast differentiation and bone density(P<0.05).Conclusion:Wenshen Zhuanggu Formula improves the degree of bone destruction induced by letrozole in rats through up regulation of the ratio of second messager cAMP to cGMP,thyroid function,and ovarian function.