本文選題：脊髓損傷　切入點(diǎn)：干細(xì)胞移植　出處：《新疆醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討骨髓間充質(zhì)干細(xì)胞(Bone mesenchymal stemcells, BMSCs)移植聯(lián)合減重步行訓(xùn)練對(duì)脊髓損傷(SCI)大鼠神經(jīng)功能恢復(fù)的影響。方法：選取Sprague-Dawley(SD)雄性大鼠,采用改良的簡(jiǎn)易打擊裝置制作T11完全性SCI模型,最終將造模成功的40只SCI大鼠根據(jù)干預(yù)方法的不同按隨機(jī)數(shù)字表法分為干細(xì)胞移植組、減重步行訓(xùn)練組、聯(lián)合治療組(干細(xì)胞移植聯(lián)合減重步行訓(xùn)練)和對(duì)照組(不做任何干預(yù)處理),每組10只大鼠。脊髓損傷手術(shù)1周后,對(duì)干細(xì)胞移植組及聯(lián)合治療組大鼠進(jìn)行干細(xì)胞移植。取傳至第3代的BMSCs,移植前一天用5-乙炔基-2'脫氧尿嘧啶核苷(5-ethynyl-2'-deoxyuridine,Edu)標(biāo)記,移植后減重步行訓(xùn)練組及聯(lián)合治療組進(jìn)行減重步行訓(xùn)練,其余2組進(jìn)行自由活動(dòng)。SCI后第1、2、3、4和5周通過(Basso-Beattie-Bresnahan, BBB)運(yùn)動(dòng)功能評(píng)分來判斷運(yùn)動(dòng)功能恢復(fù)情況,SCI第5周,應(yīng)用免疫組化染色及免疫熒光染色的方法檢測(cè)神經(jīng)特異性標(biāo)志物：神經(jīng)元特異性烯醇化酶(NSE)、微管相關(guān)蛋白(MAP-1β)、波形蛋白(VIM)陽性表達(dá),觀察移植細(xì)胞的存活、分化及損傷部位神經(jīng)纖維的恢復(fù)情況。結(jié)果：BBB運(yùn)動(dòng)功能評(píng)分顯示,SCI手術(shù)第2周聯(lián)合治療組BBB評(píng)分為(6.60±0.97)分,明顯高于其余3組,干細(xì)胞移植組和減重步行訓(xùn)練組BBB評(píng)分分別為(5.00±0.67)分、(4.80±0.63)分,均高于對(duì)照組,但兩組之間BBB運(yùn)動(dòng)功能評(píng)分差異無統(tǒng)計(jì)學(xué)意義(P0.05)。術(shù)后第3周,干細(xì)胞移植組的BBB評(píng)分為(8.00±0.670)分,高于減重步行訓(xùn)練組BBB評(píng)分為(6.80±0.79)分,(P0.05)差異具有統(tǒng)計(jì)學(xué)意義。免疫組化染色法顯示,有不同程度的神經(jīng)特異性標(biāo)志物(NSE、MAP-1β、VIM)陽性細(xì)胞充填于各組大鼠的SCI組織中,通過半定量分析法讀取各組大鼠脊髓組織中陽性表達(dá)半定量值,聯(lián)合治療組分別為(4.72±0.19)分、(4.50±0.22)分、(4.62±0.27)分,明顯高于其余3組(P0.05),組間差異具有統(tǒng)計(jì)學(xué)意義。免疫熒光染色法顯示,聯(lián)合治療組神經(jīng)特異性標(biāo)志物(NSE、MAP-1β、VIM)熒光表達(dá)明顯強(qiáng)于其余3組,同時(shí)可見明顯的神經(jīng)纖維增生、分化。結(jié)論：干細(xì)胞移植結(jié)合減重步行訓(xùn)練可有效促進(jìn)SCI大鼠神經(jīng)功能的恢復(fù),效果明顯優(yōu)于單純干細(xì)胞移植組和減重步行訓(xùn)練組。
[Abstract]:Objective: to investigate the effect of bone mesenchymal stem cells (BMSCs) transplantation combined with weight-loss walking training on the recovery of neural function in spinal cord injury (sci) rats. Finally, 40 successful SCI rats were randomly divided into stem cell transplantation group and weight-loss walking training group according to different intervention methods. The combined treatment group (stem cell transplantation combined with weight-loss walking training) and the control group (without any intervention, 10 rats in each group, 1 week after the operation of spinal cord injury), Stem cell transplantation was performed on rats in stem cell transplantation group and combined treatment group. BMSCs, which were transferred to the third generation, were labeled with 5-ethynyl-2deoxyuridine1, 5-ethynyl-2'deoxyuridine, 5-ethynyl-2'deoxyuridine1, and labeled with 5-ethynyl-2'deoxyuridine one day before transplantation. After transplantation, the weight loss walking training group and the combined treatment group were given weight-loss walking training. The other two groups were given free exercise. The recovery of motor function was evaluated by Basso-Beattie-Bresnahan (BBB) score at the 4th and 5th week after sci. Immunohistochemical staining and immunofluorescence staining were used to detect the positive expression of neuron-specific enolase (NSEN), microtubule-associated protein (MAP-1 尾) and vimentin (VIMM), and to observe the survival of transplanted cells. Results the BBB score of the combined treatment group was 6.60 鹵0.97 at the 2nd week of sci, which was significantly higher than that of the other three groups. The BBB score of stem cell transplantation group and weight-loss walking training group was 4.80 鹵0.63, which was higher than that of control group, but there was no significant difference in BBB motor function score between the two groups. The BBB score of stem cell transplantation group was 8.00 鹵0.670 at the third week after operation. The BBB score was 6.80 鹵0.79 (P 0.05) higher than that in the weight-loss walking training group (P 0.05). Immunohistochemical staining showed that there were different degrees of NSEMAP-1 尾 -VIM-positive cells filled in the SCI tissues of rats in each group. The semi-quantitative analysis method was used to read the semi-quantitative value of positive expression in spinal cord tissue of rats in each group. The score of positive expression in the combined treatment group was 4.72 鹵0.19) and 4.50 鹵0.22), which was significantly higher than that in the other three groups (P 0.05), and the difference between the two groups was statistically significant. The fluorescence expression of NSEP MAP-1 尾 -VIMM in the combined treatment group was significantly higher than that in the other three groups, and obvious nerve fiber proliferation and differentiation could be observed. Conclusion: stem cell transplantation combined with weight-loss walking training can effectively promote the recovery of nerve function in SCI rats. The effect was better than that of stem cell transplantation group and walking training group.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2
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本文編號(hào)：1619574