Department of Anatomy, College of Korean Medicine, Kyung Hee University, Seoul, 02447, Republic of Korea.
BMC Complement Med Ther. 2020 Feb 5;20(1):35. doi: 10.1186/s12906-020-2841-9.
Osteoporosis is related to the number and activity of osteoclasts. The goal of the present study was to demonstrate the effect of Chaenomelis Fructus (CF) on osteoclastogenesis and its mechanism of bone loss prevention in an OVX-induced osteoporosis model.
Osteoclasts were induced by RANKL in RAW 264.7 cells. TRAP assay was performed to measure the inhibitory effect of CF on osteoclast differentiation. Then, Expression of nuclear factor of activated T-cells (NFATc1), c-Fos which are essential transcription factors in osteoclastogenesis were detected using western blot and RT-PCR. The osteoclast-related markers were measured by RT-PCR. Moreover, the ability of CF to inhibit bone loss was researched by ovariectomized (OVX)-induced osteoporosis.
Cell experiments showed that CF inhibited osteoclast differentiation and its function. Immunoblot analyses demonstrated that CF suppressed osteoclastogenesis through the NFATc1 and c-Fos signaling pathways. RT-PCR determined that CF inhibited osteoclast-related markers, such as tartrate-resistant acid phosphatase (TRAP), cathepsin K (CTK), osteoclast-associated immunoglobulin-like receptor (OSCAR), ATPase H+ Transporting V0 Subunit D2 (ATP6v0d2) and carbonic anhydrase II (CA2). In animal experiments, CF showed an inhibitory effect on bone density reduction through OVX. Hematoxylin and eosin (H&E) staining analysis data showed that CF inhibited OVX-induced trabecular area loss. TRAP staining and immunohistochemical staining analysis data showed that CF displayed an inhibitory effect on osteoclast differentiation through NFATc1 inhibition in femoral tissue.
Based on the results of in vivo and in vitro experiments, CF inhibited the RANKL-induced osteoclasts differentiation and its function and effectively ameliorated OVX-induced osteoporosis rats.
骨质疏松症与破骨细胞的数量和活性有关。本研究旨在展示山茱萸（CF）对破骨细胞生成的影响及其在去卵巢诱导骨质疏松模型中预防骨质流失的机制。
用 RANKL 在 RAW 264.7 细胞中诱导破骨细胞。TRAP 测定法用于测定 CF 对破骨细胞分化的抑制作用。然后，通过 Western blot 和 RT-PCR 检测破骨细胞生成所必需的核因子活化 T 细胞（NFATc1）、c-Fos 的表达。通过 RT-PCR 测定破骨细胞相关标志物。此外，通过去卵巢（OVX）诱导骨质疏松研究 CF 抑制骨质流失的能力。
细胞实验表明 CF 抑制破骨细胞分化及其功能。免疫印迹分析表明 CF 通过 NFATc1 和 c-Fos 信号通路抑制破骨细胞生成。RT-PCR 确定 CF 抑制破骨细胞相关标志物，如抗酒石酸酸性磷酸酶（TRAP）、组织蛋白酶 K（CTK）、破骨细胞相关免疫球蛋白样受体（OSCAR）、ATP 酶 H+ 转运 V0 亚单位 D2（ATP6v0d2）和碳酸酐酶 II（CA2）。在动物实验中，CF 通过 OVX 显示出对骨密度降低的抑制作用。苏木精和伊红（H&E）染色分析数据表明 CF 抑制了 OVX 诱导的小梁面积丢失。TRAP 染色和免疫组织化学染色分析数据表明，CF 通过抑制 NFATc1 在股骨组织中抑制破骨细胞分化，发挥抑制作用。
基于体内和体外实验结果，CF 抑制了 RANKL 诱导的破骨细胞分化及其功能，并有效改善了 OVX 诱导的骨质疏松大鼠。
