【摘要】：目前認(rèn)為,繼發(fā)性脊髓損傷的分子機(jī)制十分復(fù)雜,且在病理過程的發(fā)生發(fā)展中起著重要作用,其中炎癥及細(xì)胞凋亡是最關(guān)鍵的影響因素,本試驗通過觀察肌肉注射低劑量雌激素與骨髓間充質(zhì)干細(xì)胞局部移植共同應(yīng)用對大鼠脊髓損傷后神經(jīng)運(yùn)動功能恢復(fù)及局部炎癥因子變化的影響。骨髓間充質(zhì)干細(xì)胞在某些條件下可定向誘導(dǎo)分化為神經(jīng)元樣細(xì)胞、軟骨細(xì)胞、脂肪細(xì)胞、血管內(nèi)皮細(xì)胞等,目前,骨髓間充質(zhì)干細(xì)胞或脂肪干細(xì)胞通過各種途徑移植治療急性脊髓損傷的效果己經(jīng)在較多的基礎(chǔ)動物實驗和一些臨床治療中獲得肯定。雌激素目前已被作為一種抗炎、抗氧化、抑制神經(jīng)細(xì)胞凋亡的生理激素用于實驗當(dāng)中,避免了大劑量糖皮質(zhì)激素沖擊治療脊髓損傷帶來的諸多副作用。研究中我們將100只雄性SD大鼠隨機(jī)分5組,假手術(shù)組僅暴露脊髓,不做損傷處理;模型組、雌激素組、干細(xì)胞組、聯(lián)合組采用改良的Allen's法建立脊髓損傷模型。干細(xì)胞組、聯(lián)合組造模后立即在損傷部位移植骨髓間充質(zhì)干細(xì)胞;雌激素組、聯(lián)合組造模后第1小時肌肉注射17β-雌二醇。造模后第1,3,5,7天,進(jìn)行BBB行為學(xué)評價;造模后6,12,24,72 h,應(yīng)用ELISA法測定脊髓勻漿組織白細(xì)胞介素1β、腫瘤壞死因子α水平,TUNEL染色觀察脊髓組織切片神經(jīng)細(xì)胞凋亡情況。實驗結(jié)果顯示:(1)與假手術(shù)組比較,模型組造模后不同時間點(diǎn)的BBB行為學(xué)評分下降(P0.05);與模型組比較,干細(xì)胞、雌激素組、聯(lián)合組造模后5,7 d的BBB行為學(xué)評分升高(P0.05),且聯(lián)合組高于干細(xì)胞、雌激素組(P0.05);(2)與假手術(shù)組比較,模型組造模后6,12,24 h的白細(xì)胞介素1β、腫瘤壞死因子α水平升高(P0.05);與模型組比較,干細(xì)胞、雌激素組、聯(lián)合組造模后12,24 h的白細(xì)胞介素1β、腫瘤壞死因子α水平降低(P0.05),且聯(lián)合組低于干細(xì)胞、雌激素組(P0.05);(3)造模后72 h,模型組神經(jīng)細(xì)胞凋亡率高于假手術(shù)組,干細(xì)胞、雌激素組、聯(lián)合組低于模型組(P0.05),且以聯(lián)合組最低(P0.05)。雌激素與骨髓間充質(zhì)干細(xì)胞聯(lián)合應(yīng)用治療脊髓損傷具有協(xié)同作用,能夠更有效抑制神經(jīng)細(xì)胞凋亡,減輕炎癥反應(yīng),促進(jìn)大鼠脊髓功能恢復(fù)。
[Abstract]:At present, it is believed that the molecular mechanism of secondary spinal cord injury is very complex and plays an important role in the pathogenesis of spinal cord injury. Inflammation and apoptosis are the most important influencing factors. The effects of intramuscular injection of low dose estrogen and local transplantation of bone marrow mesenchymal stem cells on the recovery of neural motor function and the changes of local inflammatory factors after spinal cord injury in rats were observed. Bone marrow mesenchymal stem cells can be induced to differentiate into neuron-like cells, chondrocytes, adipocytes and vascular endothelial cells under certain conditions. The effect of transplantation of bone marrow mesenchymal stem cells or adipose stem cells in the treatment of acute spinal cord injury has been confirmed in many basic animal experiments and some clinical treatments. Estrogen has been used as an anti-inflammatory, anti-oxidative, anti-neuronal apoptosis physiological hormone in the experiment, to avoid a large dose of glucocorticoid shock treatment of spinal cord injury caused by many side effects. In the study, 100 male Sprague-Dawley rats were randomly divided into 5 groups: sham-operated group only exposed spinal cord without injury treatment; Model group, estrogen group, stem cell group, combined group were used to establish spinal cord injury model by modified Allen's method. In the stem cell group, bone marrow mesenchymal stem cells were transplanted at the injured site immediately after modeling in the combined group, and 17 尾 -estradiol was injected intramuscularly in the estrogen group at 1 hour after modeling. The behavior of BBB was evaluated on the 7th day after establishment of the model, and the apoptosis of neurons in spinal cord tissue sections was observed by ELISA assay and Tunel staining. The results showed that: (1) compared with the sham operation group, the BBB behavioral score of the model group decreased at different time points after modeling (P0.05); compared with the model group, stem cells, estrogen group, The BBB behavioral score of the combined group was significantly higher than that of the stem cell group (P0.05), and the level of IL-1 尾 and TNF- 偽 in the estrogen group (P0.05); (2) was significantly higher than that in the sham operation group (P0.05) at 24 h after modeling (P0.05), and was significantly higher than that in the model group (P0.05), and compared with the model group (P0.05), the levels of IL-1 尾 and TNF- 偽 in the model group were significantly higher than those in the sham operation group (P0.05). The levels of IL-1 尾 and TNF- 偽 in stem cells, estrogen group and combination group decreased at 24 h after modeling (P0.05), and were lower in combination group than those in stem cell group. The apoptosis rate of neural cells in model group (P0.05); (3) was higher than that in sham operation group (P 0.05). Estrogen group, the combined group was lower than the model group (P0.05), and the combined group was the lowest (P0.05). Combined use of estrogen and bone marrow mesenchymal stem cells in the treatment of spinal cord injury has a synergistic effect, which can more effectively inhibit neuronal apoptosis, alleviate inflammatory reaction and promote the recovery of spinal cord function in rats.
【學(xué)位授予單位】：河北北方學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R651.2

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