摘要
背景:激素性股骨头坏死的发病机制尚不明确,该病所伴随的塌陷破坏和重建性修复或与"骨形成-吸收"偶联失效所引起微观骨结构改变有关。活血祛瘀法经临床证实对该病有积极作用。但目前针对活血祛瘀法促进骨坏死修复的作用机制尚不清楚,相关国内外文献报道较少。目的:制备激素性股骨头坏死兔模型,探究活血祛瘀法对激素性股骨头坏死修复的干预作用。方法:50只新西兰兔随机分为3组:正常组10只、模型组和中药组(桃红四物汤组)各20只,采用内毒素联合甲强龙的方法制备激素性股骨头坏死模型,最后一次肌肉注射甲强龙后24 h,中药组给予0.3 g/kg桃红四物汤混悬液灌胃,正常组和模型组灌服等体积超纯水,连续8周。实验第8周分别进行高分辨率MRI检查和病理学组织检测以评估建模效果,采用Western Blot检测兔股骨头ABCB1、RUNX2、OPN、RANK、RANKL、PPAR、骨保护素、血管内皮生长因子的蛋白表达水平。结果和结论:(1)经苏木精-伊红染色显示:模型组可见部分骨小梁断裂,骨细胞核固缩,骨小梁空骨陷窝,骨髓细胞坏死,与正常组相比空骨陷窝率差异有显著性意义(P<0.05),中药组可见成骨活动仍较为活跃,骨小梁形态较好,说明活血祛瘀法可改善坏死区域组织修复。(2)高分辨率MRI显示:模型组股骨头外形基本正常,股骨头内可见低信号带;中药组股骨头外形趋于正常,股骨头内近骨骺处有小块低密度区,大部分骨组织灰度值正常,提示活血祛瘀法可改善坏死区域影像学改变。(3)Western Blot检测结果:模型组与正常组相比RUNX2、RANK、RANKL的蛋白表达升高,ABCB1、骨保护素、血管内皮生长因子蛋白表达下降;中药组与模型组相比,除RANK、RANKL表达下降外,其余均升高,差异有显著性意义(P<0.05)。(4)结果说明:活血祛瘀法促进兔激素性股骨头坏死的修复可能与调节ABCB1、RUNX2、RANK、RANKL、骨保护素、血管内皮生长因子蛋白的表达水平有关。
BACKGROUND： The pathogenesis of steroid-induced osteonecrosis of the femoral head remains unclear. Femoral head reconstruction after collapse is related to the bone remodeling due to disorder of the bone formation-absorption coupling. ＂Removing Blood Stasis＂ Method has been shown to have a positive effect on the disease in clinics. However, the mechanisms by which the ＂Removing Blood Stasis＂ Method confers bone repair after osteonecrosis remain poorly understood. OBJECTIVE： To explore the effect of ＂Removing Blood Stasis＂ Method on bone repair in rabbits with steroid-induced femoral head osteonecrosis. METHODS： Fifty New Zealand rabbits were randomly assigned into three groups： normal control（n=10）, model（n=20） and Taohong Siwu Decoction（n=20） groups, respectively. The steroid-induced osteonecrosis of the femoral head animal models were established by intramuscular injection of endotoxin combined with methylprednisolone（MPS）. Rabbits in Taohong Siwu Decoction group were intragastrically administered with 0.3 g/kg of Taohong Siwu Decoction suspension after the last injection of methylprednisolone. The control and model groups were administrated by equal volume of ultrapure water for 8 consecutive weeks. High-resolution MRI and pathological determinations were used to assess the successful models. Protein expression levels of ABCB1, RUNX2, OPN, RANK, RANKL, PPAR, osteoprotegerin（OPG）, vascular endothelial growth factor（VEGF） in rabbit femoral heads were detected by western blot assay. RESULTS AND CONCLUSION：（1） Hematoxylin-eosin staining showed that trabecular bone fracture, karyopyknosis, empty lacunae, necrotic bone marrow cells were seen in the model group. Empty lacunae rate in the model group was significantly different from that in the normal control group（P〈0.05）. Bone formation was active and kept better trabecular morphology in the Taohong Siwu Decoction group. It suggested that ＂Removing Blood Stasis＂ Method can improve repair of necrotic area.（2） In the model group, femoral head with a wide range of low-density areas in high-resolution MRI; Taohong Siwu Decoction group showed the relatively normal shape of the femoral head with small areas of low density. It suggested that ＂Removing Blood Stasis＂ Method can improve image changes in the necrotic area.（3） Protein expression levels of RUNX2, RANK, RANKL were significantly up-regulated but protein expression levels of ABCB1, OPG, VEGF were significantly down-regulated in the model group compared with the normal control group; whereas, they were all significantly up-regulated in the Taohong Siwu Decoction group compared with the model group, except for RANK and RANKL（P〈0.05）. These results suggest that the ＂Removing Blood Stasis＂ Method can promote bone repair in steroid-induced femoral head osteonecrosis rabbits by regulating protein expressions of ABCB1, RUNX2, RANK, RANKL, OPG, and VEGF.
出处
《中国组织工程研究》
CAS
北大核心
2016年第15期2148-2155,共8页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金资助项目(81302990)
广东省杰出青年科学基金(2015A030306037)
广东省高等学校学科与专业建设专项资金项目(2013LYM 0010)~~
