Li Feimeng, Li Qihuo, Huang Xiaoqing, Wang Yunting, Ge Chana, Qi Yong, Guo Wei, Sun Hongtao
Guangdong Traditional Medical and Sports Injury Rehabilitation Research Institute, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong 510317, P.R. China.
Fourth Department of Orthopedics, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2385-2391. doi: 10.3892/etm.2017.4771. Epub 2017 Jul 11.
Osteoporosis is a systemic skeletal disease that leads to increased bone fragility and susceptibility to fracture. Approximately 50% of postmenopausal women develop osteoporosis as a result of postmenopausal estrogen deficiency. To reduce fractures related to osteoporosis in women, previous studies have focused on therapeutic strategies that aim to increase bone formation or decrease bone resorption. However, pharmacological agents that aim to improve bone fracture susceptibility exhibit side effects. Current studies are investigating natural alternatives that possess the benefits of selective estrogen receptor modulators (SERMs) without the adverse effects. Recent studies have indicated that phytoestrogen may be an ideal natural SERM for the treatment of osteoporosis. In Chinese herbal medicine, psoralen, as the predominant substance of , is considered to be a phytoestrogen and is used as a remedy for osteoporosis. A number of studies have demonstrated the efficacy of psoralen in bone formation. However, the pathways and underlying molecular mechanisms that participate in psoralen-induced osteoblast formation are not well understood. In the present study, hFOB1.19 cells were treated with psoralen at different concentrations (0, 5, 10, 15 and 20 µM) for 0, 24, 36, 48 and 72 h, respectively. Reverse transcription-quantitative polymerase chain reaction and western blot assays were performed to detect glucose transporter 3 (GLUT3) expression. A cell counting kit-8 assay was used to analyze cell proliferation. In addition the effects of mitogen activated protein kinase inhibitors on extracellular signal-regulated kinase (ERK), phosphorylated (p)-ERK, p38, p-p38, c-Jun N-terminal kinase (JNK) and p-JNK expressions and cell proliferation were measured, as was the effect of nuclear factor (NF)-κB inhibitor on P65 and GLUT3 expressions and cell proliferation. The results indicated that psoralen stimulates hFOB1.19 cell proliferation in a dose-dependent manner (P<0.05). Phospho-ERK, p38 and JNK were markedly increased by psoralen compared with the control group (P<0.05), and the specific inhibitors of ERK (SCH772984), p38 (SB203580) and JNK (SP600125) reversed the stimulatory effects of psoralen on signal marker phosphorylation (P<0.05). The rate of psoralen-induced cell proliferation was significantly suppressed by inhibitors of ERK, JNK and p38 compared with psoralen treatment alone (P<0.05). In addition, psoralen stimulated osteoblast proliferation via the NF-κB signaling pathway. Therefore, the present findings suggest that psoralen may be a potential natural alternative to SERMs in the treatment of osteoporosis and fractures.
骨质疏松症是一种全身性骨骼疾病，会导致骨脆性增加和骨折易感性增强。大约50%的绝经后女性因绝经后雌激素缺乏而患上骨质疏松症。为了减少女性与骨质疏松症相关的骨折，以往的研究集中在旨在增加骨形成或减少骨吸收的治疗策略上。然而，旨在改善骨折易感性的药物制剂存在副作用。目前的研究正在探索具有选择性雌激素受体调节剂（SERM）益处且无不良反应的天然替代物。最近的研究表明，植物雌激素可能是治疗骨质疏松症的理想天然SERM。在中药中，补骨脂素作为[此处原文缺失相关内容]的主要成分，被认为是一种植物雌激素，并被用作治疗骨质疏松症的药物。多项研究已证明补骨脂素在骨形成方面的功效。然而，参与补骨脂素诱导成骨细胞形成的途径和潜在分子机制尚未完全明确。在本研究中，分别用不同浓度（0、5、10、15和20μM）的补骨脂素处理hFOB1.19细胞0、24、36、48和72小时。进行逆转录-定量聚合酶链反应和蛋白质免疫印迹分析以检测葡萄糖转运蛋白3（GLUT3）的表达。使用细胞计数试剂盒-8分析来分析细胞增殖。此外，检测了丝裂原活化蛋白激酶抑制剂对细胞外信号调节激酶（ERK）、磷酸化（p）-ERK、p38、p-p38、c-Jun氨基末端激酶（JNK）和p-JNK表达以及细胞增殖的影响，以及核因子（NF）-κB抑制剂对P65和GLUT3表达以及细胞增殖的影响。结果表明，补骨脂素以剂量依赖性方式刺激hFOB1.19细胞增殖（P<0.05）。与对照组相比，补骨脂素显著增加了磷酸化ERK、p38和JNK的水平（P<0.05），而ERK（SCH772984）、p38（SB203580）和JNK（SP600125）的特异性抑制剂逆转了补骨脂素对信号标志物磷酸化的刺激作用（P<0.05）。与单独使用补骨脂素处理相比，ERK、JNK和p38的抑制剂显著抑制了补骨脂素诱导的细胞增殖速率（P<0.05）。此外，补骨脂素通过NF-κB信号通路刺激成骨细胞增殖。因此，本研究结果表明，补骨脂素可能是治疗骨质疏松症和骨折的一种潜在天然SERM替代物。
