基于Wnt通路的补肾养血法联合DKK1-Ab治疗对缺血型骨不连大鼠骨形成的影响  

Effect of Combination Therapy of Chinese Kidney Reinforcing and Blood Nourishing Herbal Medicine and DKK1-Ab on Ischemic Bone Union by Regulator of Protein Signaling in Vivo by Wnt Pathway

作　　者：陈潮锋 李悦[3] 直彦亮[1] 姜自伟[3] 石宇雄[1] 梁锦成[1] 刘锋[1] 何德利 CHEN Chaofeng;LI Yue;ZHI Yanliang;JIANG Ziwei;SHI Yuxiong;LIANG Jincheng;LIU Feng;HE Deli(Panyu Hospital of Chinese Medicine,Guangzhou 511400,China;Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州市番禺区中医院(广州中医药大学番禺中医院),广东广州511400 [2]广州中医药大学,广东广州510405 [3]广州中医药大学第一附属医院,广东广州510405

出　　处：《中国医药指南》2021年第14期17-20,共4页Guide of China Medicine

基　　金：广东省中医药局科研课题资助项目(20172102)。

摘　　要：目的探究基于Wnt通路的补肾养血法联合DKK1-Ab治疗对缺血型骨不连大鼠骨形成的治疗效果和作用机制。方法选取SPF级雌性SD大鼠48只,将其分为骨不连对照组、DKK1-Ab治疗组、补肾养血中药治疗组及联合治疗组。制备缺血型骨不连模型。每日给予骨不连对照组生理盐水灌胃,DKK1-Ab治疗组予以皮下注射,补肾养血中药治疗组予以中药灌胃,联合治疗组予以联合用药治疗。于第14、28日分次取材,经HE染色后观察骨组织的成骨细胞、破骨细胞、血管数量。通过酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测量血清中VEGF、BMP-2等含量,对股骨样品进行Western blot法检测β-catenin蛋白的表达水平。结果用药14 d后,与对照组相比,补肾养血中药治疗组,DKK1-Ab治疗组、联合治疗组成骨细胞数量均显著升高(P<0.05)。干预28 d后,与骨不连对照组比较,各组成骨细胞和血管数含量持续显著升高(P<0.05)。分别干预14、28 d后,与骨不连对照组比较,补肾养血中药治疗组,DKK1-Ab治疗组、联合治疗的BMP-2、VEGF蛋白表达量均增高,差异有统计学意义(P<0.05)。结论补肾养血中药能够通过改善血管血液供应促进局部骨的愈合,刺激成骨细胞生成,抑制破骨细胞分化,加快血管的形成,从而加速缺血型骨不连模型的愈合,对大鼠骨折不愈合具有良好的治疗作用。Objective To investigate the effect of combination therapy of Chinese kidney reinforcing and blood nourishing herbal medicine and DKK1-Ab on bone ununion in rats and its mechanism.Methods A total of 48 SD female rats were divided into 4 groups:control group,DKK1-Ab-treated group,Chinese kidney tonifying herbal medicine-treated group,and combination group.A model of ischemic nonunion was prepared.The nonunion control group was received normal saline gavage every day,the DKK1-Ab treatment group was received subcutaneous injection,the kidney nourishing Chinese medicine treatment group was received traditional Chinese medicine gavage,and the combination treatment group was received combined treatment.After 14,28 days,executing animal and taking material.HE staining observation:osteoblast,osteoclast,blood vessel.ELISA were used to inquiry the expression of VEGF and BMP-2 in serum and Western blot analysis were used to detect the expression ofβ-catenin in bone.Results After 14 days of treatment,compared with the control group,the number of bone cells in the kidney nourishing treatment group,the DKK1-Ab treatment group,and the combined treatment group were significantly increased(P<0.05).After 28 days of intervention,compared with the nonunion control group,the number of bone cells and blood vessels in each component continued to increase significantly(P<0.05).After 14 and 28 days of intervention,respectively,compared with the nonunion control group,the BMP-2 and VEGF protein expressions in the kidney nourishing Chinese medicine treatment group,the DKK1-Ab treatment group,and the combination treatment were all increased,and the difference was statistically significant(P<0.05)Conclusion The Chinese medicine for nourishing the kidney and nourishing blood can promote the healing of local bones by improving the blood supply of blood vessels,stimulating the production of osteoblasts,inhibiting the differentiation of osteoclasts,and accelerating the formation of blood vessels,thereby accelerating the healing of the ischemic

关 键 词：骨不连 大鼠 中医中药 补肾养血 DKK1单克隆抗体 动物实验 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R683[医药卫生—基础医学]