本文關(guān)鍵詞：補腎活血湯促進(jìn)肱骨干骨折愈合及BMSCs遷移的實驗研究　出處：《廣州中醫(yī)藥大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

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【摘要】：目的:運用補腎活血湯治療肱骨干骨折,測定相關(guān)臨床指標(biāo)衡量其臨床療效,為臨床應(yīng)用提供理論依據(jù)。以SD大鼠骨髓間充質(zhì)干細(xì)胞(bone marrow mesenchymal cells,BMSCs)為研究對象,探討補腎活血湯含藥血清是否能促進(jìn)大鼠BMSCs的增殖、遷移以及可能的相關(guān)分子機制,初步探索補腎活血湯治療骨折的生物機制,努力實現(xiàn)中醫(yī)藥干預(yù)骨折的精準(zhǔn)治療。方法:臨床觀察:選取廣州中醫(yī)藥大學(xué)第三附屬醫(yī)院住院部腎虛血瘀型的肱骨干骨折(均采用手術(shù)治療)患者40例,隨機分為治療組和對照組,其中治療組術(shù)后給予口服中藥,對照組不予中藥,余治療相同。通過骨折臨床愈合時間和骨折端的骨痂評分來評價其最終療效。實驗研究:取SD大鼠BMSCs進(jìn)行體外分離、培養(yǎng)、擴增、分化及鑒定。制備大鼠骨折模型并灌胃后提取含藥血清,MTT法檢測補腎活血湯含藥血清對大鼠BMSCs增殖的影響,Transwell檢測其對大鼠BMSCs體外遷移能力的影響,Western blot檢測其對基質(zhì)細(xì)胞衍生因子1(SDF-1)受體CXCR4蛋白表達(dá)的影響。成果:臨床觀察:在骨折臨床愈合時間方面,治療組比對照組骨折愈合時間顯著縮短,且差異具有統(tǒng)計學(xué)意義;在骨折端骨痂生長情況方面,對照組患者骨折端骨痂生長量明顯少于治療組,差異也具有統(tǒng)計學(xué)意義。實驗研究:MTT結(jié)果發(fā)現(xiàn),補腎活血湯中、高劑量組均能促進(jìn)大鼠BMSCs的增殖,其中高劑量組促進(jìn)增殖作用最明顯,差異均具有統(tǒng)計學(xué)意義。Transwell結(jié)果顯示,補腎活血湯含藥血清能促進(jìn)大鼠BMSCs的體外遷移,其中高劑量組效果最明顯,差異具有統(tǒng)計學(xué)意義。Western blot結(jié)果表明,補腎活血湯含藥血清能顯著上調(diào)BMSCs細(xì)胞膜上CXCR4蛋白的表達(dá),且呈濃度梯度性,差異具有統(tǒng)計學(xué)意義。結(jié)論:補腎活血湯能促進(jìn)腎虛血瘀型的肱骨干骨折愈合。補腎活血湯含藥血清能促進(jìn)BMSCs的增殖、遷移能力,為骨折愈合創(chuàng)造組織條件,其相關(guān)機制可能與上調(diào)CXCR4蛋白的表達(dá)有關(guān)。
[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.

【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R274.1

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