目的 觀(guān)察右歸飲和鹽酸雷洛昔芬聯(lián)合給藥對(duì)骨質(zhì)疏松模型小鼠的治療作用，并研究其作用機(jī)制。方法 將C57BL/6雌性小鼠切除卵巢制備骨質(zhì)疏松模型，手術(shù)12周后將小鼠隨機(jī)分為4組：模型組、右歸飲（0.1 g/只）組、鹽酸雷洛昔芬（10mg/kg）組和聯(lián)合給藥（右歸飲0.1 g/只與鹽酸雷洛昔芬10 mg/kg聯(lián)合給藥）組，另設(shè)假手術(shù)組，連續(xù)7周每天ig給藥1次。給藥結(jié)束后，將動(dòng)物處死，Micro-CT測(cè)定骨密度和骨組織微形態(tài)，并測(cè)定骨形態(tài)計(jì)量指標(biāo)——骨體積分?jǐn)?shù)（BV/TV）、骨表面積/骨體積（BS/BV）、骨小梁的平均厚度（Tb.Th）、骨小梁數(shù)量（Tb.N）、骨小梁分離度（Tb.Sp）；試劑盒法測(cè)定血清中I型膠原氨基端（P1NP）、膠原羧基端（CTx）水平；分離并培養(yǎng)骨髓間充質(zhì)干細(xì)胞，體外培養(yǎng)7 d，CCK-8法測(cè)定細(xì)胞增殖率，試劑盒法檢測(cè)分化相關(guān)指標(biāo)堿性磷酸酶（ALP）水平。結(jié)果 與模型組比較，右歸飲組、鹽酸雷洛昔芬組和聯(lián)合給藥組的骨密度、BV/TV、Tb.Th、Tb.N顯著升高，BS/BV和Tb.Sp顯著降低（P<0.05）；與鹽酸雷洛昔芬組比較，聯(lián)合給藥組的BV/TV、Tb.Th、Tb.N顯著升高，BS/BV和Tb.Sp顯著降低（P<0.05）。聯(lián)合給藥組P1NP水平、細(xì)胞增殖率均顯著高于其他各組（P<0.05、0.01）。ALP水平顯著高于假手術(shù)組、模型組、鹽酸雷洛昔芬組，顯著低于右歸飲組（P<0.01）。結(jié)論 在給藥時(shí)間相同的前提下，右歸飲和鹽酸雷洛昔芬聯(lián)合給藥比單獨(dú)給藥更有效地抑制小鼠骨丟失，通過(guò)促進(jìn)細(xì)胞的增殖和分化提高骨形成能力，達(dá)到增加骨密度、治療骨質(zhì)疏松的效果。

Objective To observe the effect of Yougui Drink (YGD) and raloxifene hydrochloride (RH) combined administration on mice with osteoporosis for investigating the mechanisms, so as to provide experimental basis for the treatment of osteoporosis.Methods The osteoporosis models were simulated by removal of the ovary in C57BL/6 mice. The mice were randomly divide into four groups after 12 weeks of surgery:model, YGD (0.1 g/per mouse), RH (10 mg/kg) and combined administration (YGD 0.1 g/per mouse and RH 10 mg/kg), and sham-operated group. All the mice were killed after 7 weeks of daily ig administration. Bone mineral density (BMD) and the bone microtomographic histomorphometry were assessed by Micro-CT, and the bone histomorphometry index:bone volume/total volume (BV/TV), bone surface area/bone volume (BS/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp) were detected. Next, the levels of P1NP and CTx in serum were assessed. Finally, the isolated bone mesenchymal stem cells (BMSCs) were cultured for 7 d in vitro for assessing the cell proliferation by CCK-8 method and the ability of osteogenic differentiation by quantative analysis of alkaline phosphatase (ALP).Results Compared with model group, BMD, BV/TV, Tb.Th and Tb.N of YGD group, RH group and combined administration group significantly increased, BS/BV and Tb.Sp decreased significantly (P< 0.05); Compared with RH group, BV/TV, Tb.Th, Tb.N in combined treatment group increased significantly, BS/BV and Tb.Sp decreased significantly (P< 0.05). The P1NP level and cell proliferation rate of the combined drug group were much higher than those in the other groups (P< 0.05, 0.01). The ALP level was much higher than that in sham group, model group and RH group,but much lower than that of YGD group (P< 0.01).Conclusion YGD combined with RH were more effective to attenuate bone loss of the mice than single dose of YGD and RH, by promoting cell proliferation and differentiation to improve the ability of bone formation, thus increasing the bone density and the treatment effect of osteoporosis.

國(guó)家自然科學(xué)基金資助項(xiàng)目（81460734）