Page 93 - 2019年8月第30卷第16期

Effects of Ligustri lucidi Fructus Extract on the Differentiation of Osteoclasts and the Proliferation of
Osteoblasts
LEI Lijuan 1，2，3 ，YANG Xiaoqin 1，2，3 ，ZHOU Ying 2，3，4 ，WANG Huijuan ，CHEN Lizhen ，FENG Tingting 2，3，4
2，3
2，3
（1. School of Pharmaceutical Science，Guizhou University，Guiyang 550025，China；2. Research and Development
Center for Medicinal and Edible Plant Resources of Guizhou，Guiyang 550025，China；3. Medicinal and Edible
Plant Resources Application and Development Engineering Laboratory of Guizhou，Guiyang 550025，China；
4. Pharmacy School，Guiyang College of Traditional Chinese Medicine，Guiyang 550025，China）

ABSTRACT OBJECTIVE：To investigate the effects of Ligustri lucidi Fructus extract on the differentiation of osteoclasts and the
proliferation of osteoblasts. METHODS：Rabbit primary bone marrow cells were induced with 1a，25-Dihydroxyvitamin D3 to
obtain osteoclasts. After treated with low-dose，medium-dose and high-dose of L. lucidi Fructus ethanol and water extract（both 2，
20，200 mg/L），TRACP activity determined method was used to detect the activity of TRACP in cell lysis solution and the number
of TRACP positive cells. The number of celluar bone resorption lacunae and the percentage of bone lacunae area were detected by
toluidine blue staining. Osteoblasts were obtained by L-ascorbic acid，β-sodium glycerophosphate and dexamethasone-induced
subcloning 14 of cranial parietal anterior osteocytes in mice. MTT assay and APK activity assay were used to detect relative
proliferation rate of cells and the activity of APK. RESULTS：After treated with different doses of L. lucidi Fructus extract，the
number of TRACP positive cells，the number of bone resorption lacunae and their area were changed in varying degrees. TRACP
activity，the number of its positive cells，the number of bone resorption lacunae and the percentage of bone lacunae area in
different dosage groups of L. lucidi Fructus ethanol extract and high-dose of water extract；the TRACP activity，the number of its
positive cells and the percentage of bone lacunae area in L. lucidi Fructus water extract medium-dose group were decreased
significantly，while the number of bone resorption lacunae in L. lucidi Fructus water extract low-dose group was increased
significantly（P＜0.05 or P＜0.01）. The relative proliferation rate of cells in L. lucidi Fructus extract low-dose and medium-dose
groups and APK activity of cells in extract groups were significantly increased，while the relative proliferation rate of cells were
decreased significantly in L. lucidi Fructus ethanol extract and water extract high-dose groups （P＜0.01）. CONCLUSIONS：L.
lucidi Fructus extract can inhibit TRACP activity of osteoclasts，change the bone resorption function of osteoclasts and the
proliferation behavior of osteoblasts and increase APK activity of osteoclasts.
KEYWORDS Ligustri lucidi Fructus； Ethanol extract； Water decoction； Osteoclasts； Osteoblasts； Bone resorption；
Differentiation；Proliferation

女贞子（Ligustri lucidi Fructus）为木犀科植物女贞 （M-CSF）共同刺激后分化而成，其异常分化时可造成骨
（Ligustrum lucidum Ait.）的干燥成熟果实，含有女贞子 吸收过度而导致骨量减少，进而引发OP 。成骨细胞则
[5]
苷、洋橄榄苦苷、齐墩果酸、4-羟基-β-苯乙基-β-D-葡萄糖 由间充质干细胞发育而成，可分泌细胞外基质，在骨微
[1]
苷、桦木醇等活性成分 。该药材性凉，味甘、苦，归肝、 环境下矿化为骨基质，并进一步分化成熟形成骨细胞，
肾经，具有滋补肝肾、明目乌发之功效，可用于治疗眩晕 主要参与骨的形成以及骨损伤的修复过程 [6-7] 。由此可
耳鸣、腰膝酸软、须发早白、目暗不明等症，其主要药理 见，调节破骨细胞的分化和成骨细胞的增殖可作为治疗
作用包括抗肿瘤、抗骨质疏松（OP）、抗氧化、抗衰老、增 OP 的手段之一。鉴于此，本研究以 1α，25-二羟基维生
强免疫力、升高白细胞、保肝强心等 [1-2] 。其中，抗肿瘤、 素D3[1α，25-（OH） 2VitD3]诱导兔原代骨髓细胞获得破骨
抗 OP 作用是该药材近几年研究的热点之一。中医认 细胞，以L-抗坏血酸、β-甘油磷酸钠、地塞米松诱导小鼠
为，骨由肾主导，治疗OP应从补益肝肾、强腰健膝、益气 颅顶前成骨细胞亚克隆 14 细胞（MC3T3-E1Subclone
[3]
养血、活血化瘀等方面入手 。因此，女贞子及其活性成 14）获得成骨细胞，并在此基础上初步探讨女贞子提取
分具有被开发为抗OP药物的潜力。 物对破骨细胞分化和成骨细胞增殖的影响，以期为女贞
OP 是以骨量减少，骨微结构破坏，骨强度下降，松 子防治OP提供理论依据。
质骨骨小梁变细、断裂、数量减少，皮质骨多孔和变薄导 1 材料
致骨脆性增加，容易发生骨折及骨痛为特征的全身性骨 1.1 仪器
代谢疾病，其发病机制与破骨细胞分化增强以及成骨细 3111 型 CO2 培养箱、1510 型酶标仪（美国 Thermo
胞增殖减弱有关 。破骨细胞在维持骨代谢平衡过程中 Fisher Scientific公司）；XD20-RFL型倒置荧光相差显微
[4]
可发挥重要作用，该细胞由骨髓细胞经核因子κB受体活 镜（宁波舜宇仪器有限公司）；LGJ-12 型冷冻干燥机（北
化 因 子 配 体（RANKL）和 巨 噬 细 胞 集 落 刺 激 因 子 京松源华兴科技发展有限公司）；RE-52AA 型旋转蒸发

·2248 · China Pharmacy 2019 Vol. 30 No. 16 中国药房 2019年第30卷第16期