Chen Yi-Xuan, Zhu Dao-Yu, Xu Zheng-Liang, Yin Jun-Hui, Yu Xiao-Wei, Mei Jiong, Gao You-Shui, Zhang Chang-Qing
Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Institute of Microsurgery on Extremities, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Cell Physiol Biochem. 2017;42(6):2391-2403. doi: 10.1159/000480181. Epub 2017 Aug 21.
Alcohol abuse is known to be a leading risk factor for atraumatic osteonecrosis of the femoral head (ONFH), in which the suppression of osteogenesis plays a critical role. Cordycepin benefits bone metabolism; however, there has been no study to determine its effect on osteonecrosis.
Human bone mesenchymal stem cells (hBMSCs) were identified by multi-lineage differentiation. Alkaline phosphatase (ALP) activity, RT-PCR, western blots, immunofluorescent assay and Alizarin red staining of BMSCs were evaluated. A rat model of alcohol-induced ONFH was established to investigate the protective role of cordycepin against ethanol. Hematoxylin & eosin (H&E) staining and micro-computerized tomography (micro-CT) were performed to observe ONFH. Apoptosis was assessed by TdT-mediated dUTP nick end labeling (TUNEL). Immunohistochemical staining was carried out to detect OCN and COL1.
Ethanol significantly suppressed ALP activity, decreased gene expression of OCN and BMP2, lowered levels of RUNX2 protein, and reduced immunofluorescence staining of OCN and COL1 and calcium formation of hBMSCs. However, these inhibitory effects were attenuated by cordycepin co-treatment at concentrations of 1 and 10 µg/mL Moreover, it was revealed that the osteo-protective effect of cordycepin was associated with modulation of the Wnt/β-catenin pathway. In vivo, by micro-CT, TUNEL and immunohistochemical staining of OCN and COL1, we found that cordycepin administration prevented alcohol-induced ONFH.
Cordycepin treatment to enhance osteogenesis may be considered a potential therapeutic approach to prevent the development of alcohol-induced ONFH.
酗酒是已知的非创伤性股骨头坏死（ONFH）的主要危险因素，其中成骨抑制起关键作用。虫草素对骨代谢有益；然而，尚无研究确定其对骨坏死的影响。
通过多谱系分化鉴定人骨髓间充质干细胞（hBMSCs）。评估BMSCs的碱性磷酸酶（ALP）活性、逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法、免疫荧光测定法和茜素红染色。建立酒精诱导的ONFH大鼠模型，以研究虫草素对乙醇的保护作用。进行苏木精-伊红（H&E）染色和微型计算机断层扫描（micro-CT）以观察ONFH。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）评估细胞凋亡。进行免疫组织化学染色以检测骨钙素（OCN）和I型胶原蛋白（COL1）。
乙醇显著抑制ALP活性，降低OCN和骨形态发生蛋白2（BMP2）的基因表达，降低RUNX2蛋白水平，并减少hBMSCs的OCN和COL1免疫荧光染色以及钙形成。然而，1和10μg/mL浓度的虫草素联合处理减弱了这些抑制作用。此外，发现虫草素的骨保护作用与Wnt/β-连环蛋白信号通路的调节有关。在体内，通过micro-CT、TUNEL以及OCN和COL1的免疫组织化学染色，我们发现给予虫草素可预防酒精诱导的ONFH。
虫草素治疗以增强成骨作用可被认为是预防酒精诱导的ONFH发展的一种潜在治疗方法。
