淫羊藿素通过抑制Hedgehog信号通路保护终板软骨细胞退变  

Icaritin protects the ICMT-induced degeneration of endplate chondrocytes via inhibiting Hedgehog signaling pathway

作　　者：徐永明[1] 徐宏光[1] 高智[1] 张晓玲[2] XU Yongming XU Hongguang GAO Zhi ZHANG Xiaoling(Department of Orthopedic Surgery, Yijishan Hospital, Wannan Medical College, Wuhu 241001, Anhui, China Key Laboratory of Orthopedic Surgery, Shanghai Institutes for Biological Sciences （SBIS） , Chinese Academy of Sciences （CAS） ＆ Shanghai Jiao Tong University School of Medicine （SJTUSM） , Shanghai 200025, China)

机构地区：[1]皖南医学院第一附属医院弋矶山医院脊柱外科 [2]中国科学院上海生命科学院骨科细胞与分子生物学实验室,上海200025

出　　处：《中国临床药理学与治疗学》2017年第4期373-380,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金(81272048);卫生部公益性行业专项基金(201002018)

摘　　要：目的:探讨淫羊藿素药物在体外力学诱导终板软骨细胞退变中的作用机制。方法:应用FX-5000T flexcell细胞应力加载系统体外建立大鼠终板软骨细胞退变模型,实验分正常对照组(NC组)、力学刺激组(ICMT组)、力学刺激+药物处理组(ICT+ICMT组)。甲苯胺蓝及鬼笔环肽染色观察力学刺激后细胞形态变化,CCK-8法和流式细胞术检测细胞增殖与凋亡,Western blot及RT-q PCR分别检测软骨细胞相关基因及蛋白水平变化。结果:大鼠终板软骨细胞力学刺激后出现退变,细胞形态由多边形被拉成长梭形,ICMT组、ICT+ICMT组细胞增殖率均高于NC组,但ICMT组与ICT+ICMT组间细胞增殖率比较无统计学差异,ICT+ICMT组细胞凋亡率较ICMT组明显改善,加入淫羊藿素药物处理后终板软骨细胞因力学刺激导致的退变程度有所缓解,Hedgehog信号通路被抑制。结论:淫羊藿素药物可通过抑制Hedgehog信号通路活性起到保护因力学诱导而引起的终板软骨细胞退变作用。AIM： To investigate the effect of icaritin on degeneration of endplate chondrocytes in- duced by intermittent cyclic mechanical tension （IC- MT） and its mechanism in vitro. METHODS： Rat endplate chondrocytes degeneration model was estab- lished by using FX-5000T flexcell stress loading sys- tem in vitro. Three groups were erected： normal control group （NC group）, ICMT group （10% inter- mittent cyclic mechanical tension , 10% ICMT, 0.5 Hz, 8 h/d）, and ICT ＋ ICMT group. The morpho- logical alteration of the cells was observed with tolui- dine blue staining and phalloidin staining. Cell pro- liferation and apoptosis were detected respectively by CCK-8 and flow cytometry. The mRNA levels of chondrocyte-associated genes were detected by qRT- PCR and protein levels were determined by Western blot. RESULTS ： Degeneration of endplate chondro- cytes was apparently observed after mechanical stim-ulation. In detail, cell morphology changed from polygon to long spindle after mechanical loading. The cell proliferation rates in ICMT group and ICT ＋ ICMT group were higher than NC group. However, there was no significant difference between ICMT group and ICT ＋ ICMT group. The apoptosis rate of ICT ＋ ICMT group was significantly lower than ICMT group. Interestingly, treatment with icariin effective- ly alleviated ICMT-induced degeneration of endplate chondrocytes, and significantly suppressed the Hedgehog signaling pathway. CONCLUSION ： Icaritin can efficiently protect the ICMT-induced cartilage degeneration by inhibiting the activity of Hedgehog signaling pathway.

关 键 词：淫羊藿素 终板软骨细胞 细胞退变 HEDGEHOG信号通路 

分 类 号：R965.2[医药卫生—药理学]