补肾活血汤促进肱骨干骨折愈合及BMSCs迁移的实验研究

发布时间：2018-01-08 19:30

本文关键词：补肾活血汤促进肱骨干骨折愈合及BMSCs迁移的实验研究　出处：《广州中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:运用补肾活血汤治疗肱骨干骨折,测定相关临床指标衡量其临床疗效,为临床应用提供理论依据。以SD大鼠骨髓间充质干细胞(bone marrow mesenchymal cells,BMSCs)为研究对象,探讨补肾活血汤含药血清是否能促进大鼠BMSCs的增殖、迁移以及可能的相关分子机制,初步探索补肾活血汤治疗骨折的生物机制,努力实现中医药干预骨折的精准治疗。方法:临床观察:选取广州中医药大学第三附属医院住院部肾虚血瘀型的肱骨干骨折(均采用手术治疗)患者40例,随机分为治疗组和对照组,其中治疗组术后给予口服中药,对照组不予中药,余治疗相同。通过骨折临床愈合时间和骨折端的骨痂评分来评价其最终疗效。实验研究:取SD大鼠BMSCs进行体外分离、培养、扩增、分化及鉴定。制备大鼠骨折模型并灌胃后提取含药血清,MTT法检测补肾活血汤含药血清对大鼠BMSCs增殖的影响,Transwell检测其对大鼠BMSCs体外迁移能力的影响,Western blot检测其对基质细胞衍生因子1(SDF-1)受体CXCR4蛋白表达的影响。成果:临床观察:在骨折临床愈合时间方面,治疗组比对照组骨折愈合时间显著缩短,且差异具有统计学意义;在骨折端骨痂生长情况方面,对照组患者骨折端骨痂生长量明显少于治疗组,差异也具有统计学意义。实验研究:MTT结果发现,补肾活血汤中、高剂量组均能促进大鼠BMSCs的增殖,其中高剂量组促进增殖作用最明显,差异均具有统计学意义。Transwell结果显示,补肾活血汤含药血清能促进大鼠BMSCs的体外迁移,其中高剂量组效果最明显,差异具有统计学意义。Western blot结果表明,补肾活血汤含药血清能显著上调BMSCs细胞膜上CXCR4蛋白的表达,且呈浓度梯度性,差异具有统计学意义。结论:补肾活血汤能促进肾虚血瘀型的肱骨干骨折愈合。补肾活血汤含药血清能促进BMSCs的增殖、迁移能力,为骨折愈合创造组织条件,其相关机制可能与上调CXCR4蛋白的表达有关。
[Abstract]:Objective: using Bushen Huoxue Decoction in the treatment of humeral shaft fractures, the determination of relevant clinical indicators to measure its clinical curative effect, provide a theoretical basis for clinical application. The SD rat bone marrow mesenchymal stem cells (bone marrow mesenchymal cells, BMSCs) as the research object, to investigate whether serum containing Bushen Huoxue Decoction can promote the proliferation of rat BMSCs the migration and the possible molecular mechanism, explore the biological mechanism of Bushen Huoxue Decoction in the treatment of fracture, and strive to achieve the accurate treatment of Chinese medicine intervention fracture. Methods: clinical observation: humeral shaft selection of inpatient department of kidney deficiency and blood stasis Third Affiliated Hospital of Guangzhou University of Chinese Medicine (fractures were treated by surgery) in 40 patients, were randomly divided into treatment group and the control group, the treatment group were treated with traditional Chinese medicine, the control group was not Chinese, more than the same treatment. To evaluate the clinical healing time of fracture and the fracture callus score The price of the final outcome. Experimental study: SD BMSCs of rats in vitro isolation, culture, amplification, differentiation and identification. The preparation of rat fracture model and after intragastric administration of extract serum, detection effect of Bushen Huoxue decoction containing serum MTT on the proliferation of rat BMSCs, Transwell detection and its effect on rat BMSCs in vitro the migration ability of Western, blot detection of stromal cell derived factor 1 (SDF-1) expression of CXCR4 receptor. Results: the clinical observation: the clinical healing time of fracture, the treatment group than in the control group, the fracture healing time was significantly shortened, and the difference was statistically significant; in fracture callus growth, control groups of patients with fracture callus growth was significantly less than the treatment group, the difference was statistically significant. MTT study results showed that Bushen Huoxue Decoction, high dose group could promote the proliferation of rat BMSCs in high dose group The most obvious promoting proliferation, there were statistically significant differences in the results of.Transwell showed that the serum containing Bushen Huoxue Decoction can promote the in vitro migration of rat BMSCs and the effect of high dose group was the most obvious, the difference was statistically significant.Western blot results showed that the serum containing Bushen Huoxue Decoction can significantly increase the expression of CXCR4 protein on the cell membrane of BMSCs, and a concentration gradient, the difference was statistically significant. Conclusion: Bushen Huoxue Decoction can promote blood stasis of the humeral shaft fracture healing. The serum containing Bushen Huoxue Decoction can promote BMSCs proliferation, migration, to create conditions for the healing of the fracture, the mechanism of expression may be associated with upregulation of CXCR4 protein.

【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R274.1

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