目的 研究淫羊藿苷联合GM6001治疗酒精性股骨头坏死大鼠的作用机制，同时探究其对骨保护素（OPG）/核因子-κB受体活化因子（RANK）/核因子-κB受体活化因子配体（RANKL）通路的调节作用。方法 大鼠随机分为对照组、模型组、淫羊藿苷组、GM6001组及联合组（淫羊藿苷＋GM6001），每组12只。采用ig给予大鼠白酒建立酒精性股骨头坏死大鼠模型，淫羊藿苷组按照60 mg/kg ig给予大鼠淫羊藿苷，GM6001组按照100 mg/kg尾iv GM6001，联合组同时给予淫羊藿苷（60 mg/kg）及GM6001（100 mg/kg），1次/d，连续8周，对照组及模型组给予生理盐水。旷场实验测定大鼠活动次数、活动总距离，抓力测定仪测定大鼠最大抓力；显微CT仪测定股骨头骨体积分数、骨小梁间距、骨小梁数目、骨小梁厚度；酶联免疫吸附法（ELISA）测定血清磷、钙水平，血清及股骨头骨形成蛋白-2（BMP-2）、转化生长因子-β（TGF-β）、碱性成纤维细胞生长因子（bFGF）水平。苏木精–伊红（HE）染色法测定股骨头组织病理学变化；实时定量聚合酶链式反应（PCR）测定股骨头OPG、RANK、RANKL mRNA水平；免疫印迹法测定股骨头OPG、RANK及RANKL蛋白水平。结果 与模型组比较，淫羊藿苷组、GM6001组、联合组活动次数、活动总距离、最大抓力、骨体积分数、骨小梁数目、骨小梁厚度，血清磷、钙、BMP-2、TGF-β、bFGF水平，股骨头BMP-2、TGF-β、bFGF水平，股骨头OPG mRNA及蛋白水平显著升高（P＜0.05）；骨小梁间距、股骨头RANK和RANKL的mRNA及蛋白水平显著降低（P＜0.05），且联合组降低更为显著（P＜0.05）。结论 淫羊藿苷联合GM6001能够显著修复大鼠酒精性股骨头坏死，缓解股骨头组织病理学改变，其机制可能与调节OPG/RANK/RANKL有关。

Objective To investigate the mechanism of icariin combined with GM6001 in treatment of alcohol-induced osteonecrosis of the femoral head in rats, and to explore its regulatory effect on OPG/RANK/RANKL system. Methods Rats were randomly divided into control group, model group, icariin group, GM6001 group, and combined group (icariin + GM6001), with 12 rats in each group. Rats in icariin group were given icariin 60 mg/kg by gavage, GM6001 group were given GM6001 100 mg/kg by tail vein, rats in combination group were given icariin 60 mg/kg and GM6001 100 mg/kg by gavage, once daily for 8 weeks. The control group and model group were given normal saline. The open field experiment was used to measure the number of activities and the total distance of activities of the rats. The maximum grasping force of the rats was measured by the grip tester. The volume fraction, trabecular spacing, trabecular number, and trabecular thickness of femoral head were measured by microCT. Serum phosphorus and calcium levels, BMP-2, TGF-β, bFGF levels in serum and femoral head were determined by enzyme-linked immunosorbent assay (ELISA).Hematoxylin and eosin (HE) staining was used to determine the histopathological changes of femoral head. Real-time quantitative polymerase chain reaction (PCR) was used to determine the mRNA levels of OPG, RANK, and RANKL in femoral head. The protein levels of OPG, RANK, and RANKL of femoral head were determined by Western blotting. Results Compared with the model group, the number of activities, total distance of activities, maximum grip, bone volume fraction, trabecular bone number, trabecular bone thickness, serum phosphorus, calcium, BMP-2, TGF-β, bFGF levels, and BMP-2, TGF-β, bFGF of serum and femoral head levels in icariin group, GM6001 group, and combined group were significantly increased. The mRNA and protein levels of OPG in femoral head were significantly increased (P < 0.05), the mRNA and protein levels of trabecular spacing, femoral head RANK and RANKL were significantly decreased (P < 0.05). And the combined group was more significant (P < 0.05). Conclusion Icariin combined with GM6001 can significantly repair alcohol-induced femoral head necrosis, and alleviate histopathological changes of femoral head in rats, and the mechanism may be related to the regulation of OPG/RANK/RANKL.

R285.5

海南省卫生健康行业科研项目(20A200025)