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【目的】探讨骨碎补总黄酮对Masquelet技术诱导膜内血管形成的影响。【方法】将72只SD大鼠随机分成4组,分别为模型组,药物高、中、低剂量组,每组18只。复制SD大鼠股骨中段临界骨缺损模型,并在骨缺损区行聚甲基丙烯酸甲酯(PMMA)骨水泥旷置诱导生物膜形成。从术后第1天开始,各药物组大鼠分别给予骨碎补总黄酮高、中、低剂量(0.44、0.22、0.11 g·kg~(-1)·d~(-1))灌胃治疗,模型组予等量生理盐水灌胃。给药6周后取材,采用苏木素—伊红(HE)染色法观察诱导膜组织病理变化,分别采用酶联免疫吸附(ELISA)法和实时荧光定量逆转录—聚合酶链反应(RT-qPCR)法检测诱导膜中转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)的蛋白及其mRNA表达。【结果】骨碎补总黄酮高剂量组诱导膜组织形成的新生血管多于其余各组。药物高、中、低剂量组诱导膜组织中TGF-β1、VEGF的蛋白及其mRNA表达水平均明显高于模型组(P<0.05),除VEGF mRNA指标外,其余指标的表达均具有剂量依赖性。【结论】骨碎补总黄酮在诱导膜形成期可促进TGF-β1及VEGF的表达,加速血管化进程,促进后期骨缺损重建。
【Objective】 To investigate the effect of total flavonoids of Rhizoma Drynariae on Masquelet induced intravascular membrane formation. 【Methods】 Seventy-two SD rats were randomly divided into 4 groups: model group, high, medium and low dose group, with 18 rats in each group. The model of the critical defect of the middle femur of SD rats was duplicated and biomembrane formation was induced by cement removals of PMMA in the bone defect area. From the first day after operation, the rats in each drug group were given gavage at high, medium and low doses (0.44,0.22,0.11 g · kg -1 · d -1) Treatment, the model group to the same amount of saline gavage. Six weeks after the administration, the pathological changes of the induced tissue were observed by hematoxylin-eosin (HE) staining, and the changes of the pathological changes were analyzed by enzyme-linked immunosorbent assay (ELISA) and real-time quantitative reverse transcription polymerase chain reaction Methods The protein and mRNA expression of TGF-β1 and VEGF were detected by ELISA. 【Result】 The results showed that there were more neovascularization than the other groups induced by high concentration of total flavonoids of Rhizoma Drynariae. The protein and mRNA expression levels of TGF-β1 and VEGF in high, medium and low dose groups were significantly higher than those in model group (P <0.05), except for VEGF mRNA, the other indexes were all dose-dependent Sex. 【Conclusion】 Rhizoma Drynariae flavonoids can promote the expression of TGF-β1 and VEGF in the induction of membrane formation, accelerate the process of vascularization and promote the reconstruction of late bone defects.