骨碎补总黄酮对Masquelet技术诱导膜内血管形成的影响

Effects of Flavonoids of Rhizoma Drynariae on Formation of Blood Vessels in Induced Membrane by Masquelet Technique

【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 explore the effects of flavonoids of Rhizoma Drynariae on the formation of blood vessels in the induced membrane by Masquelet technique. Methods Seventy-two SD rats were randomly divided into 4 groups,namely model group,and high-,middle-and low-dose drug groups,18 rats in each group. Rat model of critical-sized femoral defect was built,and then polymethyl methacrylate（PMMA） bone cement spacer was inserted into the bone defect to induce the formation of membrane. From the first day after surgery,the rats in high-,middle-and low-dose drug groups were given gastric gavage of 0.44,0.22,0.11 g·kg^-1·d^-1 of Rhizoma Drynariae flavonoids, respectively, and the rats in the model group were given the same volume of normal saline. After 6-week medication,the pathologic features of bone cement-induced membrane were observed by Haematoxilin-Eosin（HE）staining,the contents of transforming growth factor（TGF）-β1 and vascular endothelial growth factor（VEGF）proteins in the induced membrane were tested by enzyme-linked immunosorbent assay（ELISA）,and the mRNA levels of TGF-β1 and VEGF in the induced membrane were determined by real-time reverse transcription-quantitative polymerase chain reaction（RT-qPCR）. Results More blood vessels in the induced membrane of the high-dose group were found than those of the other groups under the light microscope. The protein and mRNA expression levels of TGF-β1 and VEGF in the induced membrane of the 3 drug groups were much higher than those of the model group（P 0.05）. Except for the VEGF mRNA expression level, the changes of other indexes were dose-dependent. Conclusion Flavonoids of Rhizoma Drynariae are effective on enhancing the protein and mRNA expression levels of TGF-β1 and VEGF in the induced membrane,and can accelerate the vascularization,which promotes the reconstruction of bone defect.