补肾方剂对绝经后骨质疏松症大鼠的影响及其作用机制研究  

Effects and Potential Mechanisms of Bushen Recipes on Postmenopausal Osteoporosis

作　　者：徐绍俊 黄建烽[2] 邵敏 王大天 孙友强 姜涛[2] 黄永青 

机构地区：[1]广州市正骨医院,广东广州510045 [2]广州中医药大学第三临床医学院,广东广州510360 [3]广州中医药大学第三附属医院,广东广州510240 [4]海南省中医院,海南海口570000 [5]广州中医药大学第一临床医学院,广东广州510405

出　　处：《中药新药与临床药理》2017年第5期588-593,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81373654)

摘　　要：目的考察补肾方剂六味地黄丸和金匮肾气丸对绝经后骨质疏松症(POMP)大鼠的治疗效果及其潜在的作用机制。方法采用双侧卵巢切除术构建POMP模型大鼠后,连续3个月灌胃给药治疗,通过双能X线骨密度仪和雌激素酶联免疫分析试剂盒分别检测大鼠股骨骨密度和血清雌激素水平。同时,体外分离培养大鼠原代成骨细胞,10,50,100 ng·μL^(-1)不同浓度补肾方剂处理后,通过CCK-8试剂盒和碱性磷酸酶(ALP)试剂盒分别检测成骨细胞活性和ALP活性;通过RT-PCR和WB检测成骨细胞内雌激素受体ERs(ERα,ERβ,ERRα)Collagen I和p-AMPK表达变化。结果金匮肾气丸可显著改善POMP大鼠股骨骨密度和血清雌激素水平(P<0.05),但六味地黄丸作用不明显。成骨细胞经不同浓度补肾方剂处理后,浓度为50,100 ng·μL^(-1)的金匮肾气丸可有效提高成骨细胞活力、ALP活性和Collagen I蛋白表达,且金匮肾气丸浓度越高,作用效果越明显,而六味地黄丸对成骨细胞作用不显著。此外,金匮肾气丸可显著增加ERα的表达(P<0.05,P<0.01),抑制p-AMPK蛋白表达,但对ERβ、ERRα表达影响不明显。结论金匮肾气丸可通过促进ERα表达,抑制p-AMPK有效治疗POMP,明显促进成骨细胞增殖分化,改善成骨细胞功能,但六味地黄丸作用效果不显著。Objective To study the effects of Liuwei Dihuang Pill and Jinkui Shenqi Pill on treating postmenopausal osteoporosis （PMOP） and the underlying mechanisms. Methods Rat models of PMOP were established by bilateral ovariectomy. The bone mineral density （BMD） of femur and levels of serum estrogen in PMOP rats which treated with Jinkui Shenqi Pill and Liuwei Dihuang Pill for 3 months were examined by dual energy X ray absorptiometry and estrogen detection kit, respectively. Meanwhile, the primary osteoblasts were isolated in vitro and treated with Jinkui Shenqi Pill and Liuwei Dihuang Pill with different concentrations （10, 50, 100 ng·μL^-1 ）. The cell viability and ALP activity were evaluated by CCK-8 kit and alkaline phosphatase kit; mRNA expressions of estrogen receptors （ERα, ERβ, ERRα） were examined by RT-PCR, and protein expressions of Collagen I, ERot and p-AMPK were measured by Western Blot method. Results The BMD of femur and levels of serum estrogen in POMP rats treated with Jinkui Shenqi Pill increased significantly （P 〈 0.05）, while there was no obvious change in Liuwei Dihuang Pill treated group. Meanwhile, the cell viability, ALP activity and the expressions of Collagen I of osteoblasts increased notably after treatment of 50 and 100 ng·μL^-1 Jinkui Shenqi Pill, especially at the concentration of 100 ng·μL^-1; but there were

关 键 词：金匮肾气丸 六味地黄丸 绝经后骨质疏松症 卵巢切除术 成骨细胞 

分 类 号：R285.5[医药卫生—中药学]