六味地黄汤及其“补泻”药对对绝经后骨质疏松大鼠股骨和肾脏中OPG及RANKL蛋白表达的影响

Effect of Liuwei Dihuangtang and ‘Reinforcing and Reducing’ Herb Pairs on OPG and RANKL in Femur and Kidney of Ovariectomized Rats Model with Osteoporosis

目的：观察六味地黄汤及其3个“一补一泻”药对对绝经后骨质疏松模型大鼠股骨及肾脏中骨保护素（OPG）及核转录因子-xB受体活化因子配体（RANKL）的影响。方法：将3～4月龄sD大鼠随机分为空白组（假手术组，蒸馏水10mL·kg^-1），模型组（蒸馏水10mL·kg^-1），六味地黄汤组（6．75g·kg^-1），山茱萸一牡丹皮组（1．89g·kg^-1），熟地黄一泽泻组（2．97g·kg^-1），山药．茯苓组（1．89g·kg^-1），阿仑膦酸钠组（阳性药物组，6．3g·kg^-1）。除空白组外，其余各组大鼠均采用摘除双侧卵巢的方法制备绝经后骨质疏松症大鼠模型。造模后第16天开始，各组按相应剂量连续灌胃给药90d，每周观察各组大鼠体重的变化，并采用免疫组化法检测股骨成骨细胞和肾脏肾小管上皮细胞中OPG，RANKL蛋白的表达。结果：与模型组比较，六味地黄汤组能上调股骨成骨细胞及肾脏肾小管上皮细胞中的OPG的蛋白表达、下调RANKL的蛋白表达；但山茱萸一牡丹皮药对仅能降低股骨成骨细胞中OPG的灰度值（P〈0．05），升高RANKL的灰度值（P〈0．01），即山茱萸一牡丹皮药对仅能上调股骨成骨细胞中OPG的蛋白表达、下调RANKL的蛋白表达（灰度值越低，OPG，RANKL的蛋白表达越强）。结论：六味地黄汤改善绝经后骨质疏松症的机制之一可能与其方中山茱萸一牡丹皮药对调节股骨中的OPG及RANKL的蛋白表达有关。

Objective： To investigate the effects of Liuwei Dihuangtang and its three ＇ reinforcing and reducing＇ herb pairs on the osteoprotegerin （OPG） and the nucleus factor-KB receptor activation factor ligands （RANKL） in kidney and femur of ovariectomized rat models with osteoporosis. Method： The 34 month SD rats were randomly divided into blank group （sham operation group, distilled water, 10 mL.kg^-1）, model group （distilled water, 10 mL＇kg^-1）, Liuwei Dihuangtang group （6.75 g·kg^-1）, Corni Fructus-Moutan Cortex group （ 1.89 g·kg^-1 ） , Rehmanniae Radix Praeparata,Alismatis Rhizoma group （2.97 g·kg^-1 ） , Dioscoreae Rhizoma- Poria group （ 1.89 g·kg^-1 ）, and alendronate group （ positive drug group, 6.3 g·kg^-1 ）. Except for the blank group, 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 weight changes of the rats were observed every week, and the protein expression levels of OPG and RANKL were detected by using immunohistochemical method. Result： 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 renal tubular epithelial cells was decreased by Liuwei Dihuangtang. But the gray value of OPG was reduced （P 〈 0. 05） and the RANKL in osteoblast was increased （P 〈 0.01 ） by Corni Fructus-Moutan Cortex pair, indicating that the OPG protein expression was increased and the protein expression of RANKL in osteoblast was decreased （the lower of gray value, the higher expression of OPG and RANKL）. Conclusion： 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.