【摘要】：目的利用甲強龍(MP)藥理學機制和骨髓間充質(zhì)干細胞(BMSCs)生物特點,探討甲強龍(MP)在骨髓間充質(zhì)干細胞治療大鼠脊髓損傷(SCI)中作用機制及其特點。方法取雄性SD大鼠4周齡4只,分別處死并分離股骨、脛骨獲得骨髓,在全貼壁法下培養(yǎng)并鑒定第3代BMSCs,收集備用并用Brd U染色作細胞標記。取SD大鼠6月齡40只,隨機等分為4組。A組(對照組)和B組(甲強龍治療組)、C組(BMSCs治療組)、D組(甲強龍+BMSCs聯(lián)合治療組),按上述分組采用改良Allen法制備T10脊髓損傷雄性大鼠模型,B組造模后立即尾靜脈緩慢推注甲強龍(30mg/kg),C組造模后2 h注射Brd U標記的BMSCs懸液2 m L,D組給予以上兩種干預方法。采用BBB評分法評估術(shù)后1d、3d、7d、10d、14d大鼠后肢神經(jīng)功能狀況,14d后處死各組大鼠并取出損傷部位的脊髓樣本,分別進行TNF-α、IL-1β免疫組織化學染色,Tunel染色,各組統(tǒng)計5個高倍視野內(nèi)TNF-α、IL-1β表達平均陽性細胞數(shù)并檢測細胞凋亡指數(shù);另外,對比C組和D組中BMSC存活率。用統(tǒng)計軟件SPSS20.0分析。結(jié)果術(shù)后1 d,四組大鼠后肢神經(jīng)功能較術(shù)后均無明顯提高,組間BBB評分的比較差異無統(tǒng)計學意義(P0.05)。術(shù)后3 d,B組和D組BBB評分在較術(shù)后1 d均升高(P0.05),C組與A組仍無明顯差異,與A組比較,B組和D組評分比較差異有統(tǒng)計學意義(P0.05);術(shù)后7、10、14天,四組均較前一時間點升高(P0.05),其中B,C,D組明顯高于A組,差異有統(tǒng)計學意義(P0.05);D組明顯高于B,C組,差異有統(tǒng)計學意義(P0.05);免疫組化染色顯示B組和C組的TNF-α及IL-1β平均陽性細胞數(shù)均明顯低于A組;D組明顯高于B組和C組的TNF-α及IL-1β平均陽性細胞數(shù);Brd U染色顯示C組BMSC的陽性率為明顯低于D組,兩組比較差異有統(tǒng)計學意義(P0.05)。結(jié)論1、甲強龍和BMSCs移植均可改善SCI大鼠的神經(jīng)功能恢復,以聯(lián)合作用效果最佳;2、甲強龍和BMSCs均能降低局部TNF-α、IL-1β表達水平,減少損傷部位的細胞凋亡,甲強龍作用效果強于BMSCs,二者協(xié)同作用效果明顯增強;3、甲強龍有效的提升了移植BMSCs的存活率。
[Abstract]:Objective to investigate the mechanism and characteristics of methylketene (MP) in the treatment of spinal cord injury (SCI) in rats by using the pharmacological mechanism and the biological characteristics of bone marrow mesenchymal stem cells (BMSCs). Methods four male SD rats of 4 weeks old were killed and separated from femur. Bone marrow was obtained from the tibia. The third generation of BMSCs, was cultured and identified with Brd U staining. Forty SD rats aged 6 months were randomly divided into 4 groups: group A (control group) and group B (), C group) (BMSCs group), D group). The male model of T10 spinal cord injury was established by modified Allen method. In group B, the tail vein was injected with methylenolone (Brd U labeled BMSCs suspension 2 mL) 2 hours after the model was established in group B. Group D was given the above two intervention methods. The neurological function of the hind limbs of the rats was evaluated by BBB scoring method on the 1st day, 3rd day, 7th day, 10d and 14th day after operation. After 14 days, the rats in each group were killed and the spinal cord samples of the injured area were taken out. TNF- 偽, IL-1 尾 immunohistochemical staining and Tunel staining were performed respectively. The average number of TNF- 偽 and IL-1 尾 positive cells and apoptosis index were measured in each group. In addition, the survival rate of BMSC in group C and group D was compared. Statistical software SPSS20.0 was used to analyze. Results on the 1st day after operation, the nerve function of the hind limbs of the four groups was not significantly improved than that of the control group, and the difference of BBB score between the four groups was not statistically significant (P0.05). The BBB scores in group B and group D were significantly higher than those in group A on the 3rd day after operation (P0.05), C and group A still had no significant difference, compared with group A, the scores of group B and group D were significantly higher than those of group A (P0.05); After 7 days 10 to 14 days, the four groups were higher than the previous time point (P0.05), among them, group D was significantly higher than group A (P0.05), the difference was statistically significant (P0.05) in); D group was significantly higher than that in group C (P0.05). Immunohistochemical staining showed that the average number of TNF- 偽 and IL-1 尾 positive cells in group B and C was significantly lower than that in group A, and the number of TNF- 偽 and IL-1 尾 in group D was significantly higher than that in group B and C. Brd U staining showed that the positive rate of BMSC in group C was significantly lower than that in group D, and the difference between the two groups was statistically significant (P0.05). Conclusion 1. Both methylenolone and BMSCs transplantation can improve the recovery of nerve function in SCI rats, and the combined effect is the best. (2) both methylenolone and BMSCs could decrease the expression of TNF- 偽 and IL-1 尾, and decrease the apoptosis of injured site. The effect of methylenolone was stronger than that of BMSCs,. 3. Methylenolone effectively improved the survival rate of BMSCs transplantation.
【學位授予單位】：河北北方學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R651.2

【參考文獻】

相關(guān)期刊論文 前5條

1 Fei Yin;Chunyang Meng;Rifeng Lu;Lei Li;Ying Zhang;Hao Chen;Yonggang Qin;Li Guo;;Bone marrow mesenchymal stem cells repair spinal cord ischemia/reperfusion injury by promoting axonal growth and anti-autophagy[J];Neural Regeneration Research;2014年18期

2 趙林;馮智慧;焦淑賢;李南南;;全骨髓貼壁法分離培養(yǎng)大鼠骨髓間充質(zhì)干細胞及其生物學特性[J];中國組織工程研究與臨床康復;2011年32期

3 李洪秋;王哲;阿良;;骨髓間充質(zhì)干細胞移植對大鼠脊髓損傷后氧化應(yīng)激的影響[J];脊柱外科雜志;2010年03期

4 李輝映;文益民;羅亞麗;藍旭;汪大彬;孫振剛;胡凌云;;不同次數(shù)移植BMSCs修復脊髓損傷的比較研究[J];中國修復重建外科雜志;2010年02期

5 馮大雄;鐘德君;陽運康;宋躍明;;靜脈移植骨髓間充質(zhì)干細胞對大鼠脊髓損傷后軸突再生的作用[J];中國脊柱脊髓雜志;2006年12期

，

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