补肾方联合骨膜原位成骨治疗肾虚型大鼠股骨骨缺损的机制  

Mechanism of Bushen Formula combined with periosteum osteogenesis for treatment of femoral bone defect in rats with kidney deficiency

作　　者：张严 冯俊铭 谢磊 陈柏行 曾展鹏[2] 姜自伟[2] 蔡杨庭 高怡加[4] ZHANG Yan;FENG Jun-ming;XIE Lei;CHEN Bai-hang;ZENG Zhan-peng;JIANG Zi-wei;CAI Yang-ting;GAO Yi-jia(Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Province Traditional Chinese Medical Hospital,Guangzhou 510120,China)

机构地区：[1]广州中医药大学,广州510405 [2]广州中医药大学第一附属医院,广州510405 [3]广东省中医院,广州510120 [4]广州中医药大学第一附属医院一骨科,广州510405

出　　处：《中华中医药杂志》2021年第9期5526-5531,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：广东省自然科学基金项目(No.2018A030313369);国家自然科学基金项目(No.81774337);广东省中医药局科研基金项目(No.20181080);广东省医学科学技术研究基金项目(No.B2018097)。

摘　　要：目的:探究骨膜原位成骨结合补肾方对肾虚型大鼠股骨骨缺损修复的影响及其作用机制。方法:36只雌性SD大鼠随机分为6组:自体骨膜补肾方高、中、低剂量组,自体骨膜组,胶原膜组和无骨膜组,建立肾虚型大鼠股骨大段骨缺损模型,分别予自体原位骨膜和Bio-Gide胶原膜覆盖治疗,自体骨膜补肾方组于术后分别予高、中、低剂量补肾方灌胃治疗8周。术后第12周,采用X线检查及骨组织HE染色进行Lane-Sandhu X射线评分及Lane-Sandhu组织学评分;采用实时荧光定量RT-PCR法检测骨组织中转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)、骨形态发生蛋白-2(BMP-2)和核心结合因子(Runx-2) mRNA的表达。结果:X线检查及骨组织HE染色结果显示,治疗组Lane-SandhuX射线、Lane-Sandhu组织学评分均明显高于无骨膜组(P<0.01),其中自体骨膜补肾方高、中剂量组明显高于自体骨膜组(P<0.01),且自体骨膜组明显高于胶原膜组(P<0.01,P<0.05);RT-PCR检测结果显示,与无骨膜组比较,自体骨膜补肾方各剂量组、自体骨膜组及胶原膜组的骨组织中TGF-β1、VEGF、BMP-2、Runx-2 mRNA的表达显著上调(P<0.01,P<0.05)。结论:骨膜原位成骨结合补肾方治疗能显著提高肾虚型大鼠股骨骨缺损修复效能,其作用明显优于单纯骨膜原位成骨,且与胶原膜相比较,自体骨膜具有更好的成骨和成血管能力。Objective: To investigate the effects of combination of periosteum osteogenesis(PO) and Bushen Formula on the repair of the femur defect of kidney-deficiency rats and explore its mechanism. Methods: A total of 36 female SD rats were randomly divided into 6 groups: PO with Bushen Formula group(high dose), PO with Bushen Formula group(medium dose), PO with Bushen Formula group(low dose), PO group, collagen membrane(CM) group and no periosteum(NP) group. The models for the large defect of femur of kidney-deficiency rats were established, and were respectively treated with PO and Bio-Gide collagen membrane covering. PO with Bushen Formula group were respectively given high, medium and low doses of Bushen Formula for 8 weeks after operation. 12 weeks later, Lane-Sandhu X-ray score and Lane-Sandhu histological score were performed by X-ray examination and HE staining of bone tissue. Expression of TGF-β1, VEGF, BMP-2 and Runx-2 mRNA in bone tissue were detected by RT-PCR. Results: The results showed that the histological scores of Lane-Sandhu X-ray and Lane-Sandhu in the groups with treatments were significantly higher than those in the group without periosteum(P<0.01), in which PO with Bushen Formula group was significantly higher than PO group(P<0.01), and PO group was significantly higher than CM group(P<0.01,P<0.05). RT-PCR detection showed that compared with NP group, PO with Bushen Formula group, PO group and CM group had significantly up-regulated the mRNA expression of TGF-β1、VEGF、BMP-2、Runx-2, in bone tissue(P<0.01, P<0.05).Conclusion: Periosteum osteogenesis combined with Bushen Formula can significantly improve the repair efficiency of femur defect in rats with kidney deficiency, and its effect is significantly better than that of periosteum osteogenesis alone. Compared with collagen membrane, autogenous periosteum has better osteogenesis and vascularization ability.

关 键 词：中医中药 骨膜原位成骨 肾虚模型 补肾方 骨缺损 引导骨再生 

分 类 号：R285.5[医药卫生—中药学]