壮骨止痛方调节OPG/RANKL平衡抗绝经后骨质疏松作用

Regulate Effect of Zhuanggu Zhitong Prescription on the Balance of OPG/RANKL to Anti-osteoporosis

关键词:  骨质疏松  壮骨止痛方  骨保护素  RANKL  OPG/RANKL

Key words:  osteoporosis  ZhuangguZhitong Prescription  osteoprotegerin (OPG)  receptor activator of NF-кB ligand (RANKL)  OPG/RANKL

摘要:

目的 研究壮骨止痛方对去卵巢骨质疏松大鼠骨保护素（osteoprotegerin，OPG）和NF-кB受体活化因子（receptoractivator of NF-кB，RANK）/RANK配体（RANK ligand，RANKL）的影响，探讨其抗骨质疏松的作用机制。方法 72只SD大鼠按体质量随机抽出假手术组12只，造模组60只，造模组采用双侧去卵巢法造模，假手术组只切除卵巢周围相应等量的脂肪。术后造模组大鼠按体质量随机分为模型组、壮骨止痛方高剂量组（13.2 g·kg^-1）、中剂量组（6.6 g·kg^-1）、低剂量组（3.3 g·kg^-1）和戊酸雌二醇组，每组12只。术后第5天开始药物干预，模型组和假手术组给予相应体积的纯净水，持续13周。给药结束后，股骨进行病理组织检测，酶联免疫法检测大鼠血清和骨组织OPG、RANKL含量，免疫组化法检测大鼠骨组织OPG、RANKL蛋白表达。结果 模型组大鼠股骨骨小梁面积率显著降低，血清和骨组织OPG含量显著下降，RANKL含量显著增加，OPG/RANKL比值显著降低，骨组织OPG蛋白表达显著下调，RANKL蛋白表达显著增强，OPG/RANKL比值显著降低，与假手术组比较，差异有统计学意义（P<0.05或P<0.01）。壮骨止痛方各剂量组大鼠股骨骨小梁面积率显著增加，显示其具有良好的抗骨质疏松作用；血清和骨组织OPG含量显著增加，RANKL含量显著下降，OPG/RANKL比值升高；骨组织OPG蛋白表达显著增强，RANKL蛋白表达显著下调，OPG/RANKL比值显著升高，与模型组比较，差异有统计学意义（P<0.05或P<0.01）。结论 去卵巢大鼠OPG/RANKL比值显著降低，壮骨止痛方能显著升高去卵巢大鼠OPG/RANKL比值，调节OPG/RANKL平衡是壮骨止痛方抗绝经后骨质疏松的作用机制之一。

Abstract:

OBJECTIVE To explore the effect of Zhuanggu Zhitong Prescription on the osteoprotegerin (OPG) and receptor activator of NF-кB ligand (RANKL) in ovariectomized rats and investigate its mechanism in anti-osteoporosis.METHODS The 72 SD rats were randomly divided into sham group and model group,the model group rats were ovariectomized and the sham group rats were excised equal weight fat.After the modeling,the rats were randomly divided into model group,Zhuanggu Zhitong Prescription high-dose group (13.2 g·kg^-1),middle-dose group (6.6 g·kg^-1),low-dose group (3.3 g·kg^-1) and estradiol valerate group,12 rats in each group.Five days after the operation started to drug therapy for 13 weeks.The model group and sham group rats were given equal volume water.After the test,the femur was examined by pathological examination.The content of OPG and RANKL in serum and bone tissue of rats were detected by Elisa.The protein expression of OPG and RANKL in bone tissue were detected by immunohistochemical method.RESULTS The femur trabecular area ratio of ovariectomized rats was significantly declined,the content of OPG in serum and bone tissue of model group was significantly declined,and the RANKL was significantly increased.The protein expression of OPG was significantly declined,and the protein expression of RANKL was significantly increased,compared with sham operation group,the difference was statistically significant (P<0.05,P<0.01).Zhuanggu Zhitong Prescription could significantly increase the femur trabecular area ratio,indicating a good anti-postmenopausal osteoporosis effect.The content of OPG in serum and bone tissue of Zhuanggu Zhitong Prescription groups were significantly increased,and the RANKL were significantly declined;the protein expression of OPG was significantly increased and RANKL was significantly declined,compared with the model group,the difference was statistically significant (P<0.05,P<0.01).CONCLUSION The OPG/RANKL ratio significantly decreased in ovariectomized rats,Zhuanggu Zhitong Prescription can obviously increase the ratio of OPG/RANKL in ovariectomized rats.Regulating balance of OPG/RANKL may be part of the mechanism of Zhuanggu Zhitong Prescription to anti-osteoporosis.