甲强龙在骨髓间充质干细胞治疗大鼠脊髓损伤中的作用及对肿瘤坏死因子-α和白细胞介素-1β表达的影响

发布时间：2018-12-28 10:15

【摘要】：目的探讨甲强龙(MP)在骨髓间充质干细胞(BMSCs)治疗大鼠脊髓损伤(SCI)中的作用及对局部肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β表达变化的影响。方法采用改良Allen法制备40只T10脊髓损伤雄性大鼠模型,随机数字表法随机分为对照组、MP组、BMSCs组和MP+BMSCs组4组,每组10只。按上述分组分别在脊髓损伤后立即尾静脉缓慢推注30 mg/kg MP和/或在脊髓损伤后2 h于脊髓损伤部位注射全贴壁法培养并以Brd U标记的BMSCs。术后1、7、14 d,采用BBB运动评分法评价大鼠后肢运动功能恢复。14 d后处死各组大鼠并取出脊髓样本,分别进行TNF-α、IL-1β免疫组织化学染色和Tunel染色,各组统计视野内TNF-α、IL-1β表达阳性细胞数并检测细胞凋亡率;采用Brd U单染观察BMSCs组和MP+BMSCs组中Brd U阳性率。结果术后1 d,各组BBB评分差异无统计学意义(χ2=1.0756,P=0.7829)。术后7 d,各组大鼠双后肢运动功能较前明显恢复,其中MP组(χ2=8.3265,P=0.0325)和BMSCs组(χ2=14.1166,P=0.0036)的BBB评分明显高于对照组;MP+BMSCs组的BBB评分明显高于MP组(χ2=17.7186,P=0.0002)和BMSCs组(χ2=15.8110,P=0.0024)。术后14 d,各组大鼠双后肢运动功能明显恢复,MP+BMSCs组BBB评分明显高于MP组(χ2=24.7259,P0.0001)和BMSCs组(χ2=25.6014,P0.0001),MP组(χ2=13.5060,P=0.0062)和BMSCs组(χ2=8.9613,P=0.0299)的BBB评分明显高于对照组。术后14 d,对照组、MP组、BMSCs组和MP+BMSCs组的细胞凋亡率分别为(48.47±5.70)%、(31.95±3.58)%、(41.39±2.33)%和(23.48±2.69)%,其中,MP组(q=14.84,P0.0001)和BMSCs组(q=6.716,P=0.0002)明显低于对照组,MP+BMSCs组明显低于MP组(q=7.332,P=0.0001)和BMSCs组(q=15.460,P0.0001)。术后14 d,免疫组织化学染色结果显示,MP组(q=14.710,P0.0001;q=6.710,P0.0001)和BMSCs组(q=6.502,P=0.0001;q=2.849,P=0.0514)的TNF-α及IL-1β阳性细胞数均明显低于对照组,MP组(q=5.573,P=0.0004;q=4.596,P=0.0025)和BMSCs组(q=13.780,P0.0001;q=8.456,P0.0001)的TNF-α及IL-1β阳性细胞数明显高于MP+BMSCs组。术后14 d,Brd U染色结果显示,BMSCs组的Brd U阳性率为(6.600±0.3399)%,明显低于MP+BMSCs组的(9.300±0.5175)%(t=4.361,P=0.0004)。结论 MP可明显提高BMSCs移植治疗SCI的大鼠后肢运动功能恢复,二者可协同降低局部TNF-α和IL-1β表达水平,并且MP有效提升了移植BMSCs的存活。
[Abstract]:Objective to investigate the role of methylketene (MP) in the treatment of rat spinal cord injury (SCI) with bone marrow mesenchymal stem cell (BMSCs) and its effect on the expression of local tumor necrosis factor (TNF) 伪 and interleukin (IL) 1 尾. Methods Forty male rats with T10 spinal cord injury were made by modified Allen method. The rats were randomly divided into control group, MP group, BMSCs group and MP BMSCs group with 10 rats in each group. According to the above grouping, 30 mg/kg MP of caudal vein was injected immediately after spinal cord injury and / or 2 hours after spinal cord injury, BMSCs. cultured with Brd U was injected into the site of spinal cord injury with total adherent method. The motor function of hind limbs was evaluated by BBB motor scoring method on the 1st day and 7th day after operation. After 14 days, the rats in each group were killed and the spinal cord samples were taken out. TNF- 伪, IL-1 尾 immunohistochemical staining and Tunel staining were performed respectively. The number of TNF- 伪 and IL-1 尾 positive cells and the apoptosis rate were measured in each group. The positive rate of Brd U in BMSCs group and MP BMSCs group was observed by Brd U single staining. Results on the 1st day after operation, there was no significant difference in BBB score among the groups (蠂 ~ 2 = 1.0756 / P = 0.7829). On the 7th day after operation, the motor function of the hind limbs of the rats in each group recovered obviously, and the BBB scores of the MP group (蠂 2 + 8.3265) and the BMSCs group (蠂 2 + 14. 116% P0. 0036) were significantly higher than those of the control group. The BBB score of MP BMSCs group was significantly higher than that of MP group (蠂 2 + 17.718 6 P 0. 0002) and BMSCs group (蠂 2 + 15.81 0 0. 0024). On the 14th day after operation, the BBB score of, MP BMSCs group was significantly higher than that of MP group (蠂 2 + 24.7259), BMSCs group (蠂 2 + 25.6014 P 0.0001), MP group) and BMSCs group (蠂 2 + 13.5060 + P 0. 0062) and BMSCs group (蠂 2 8.9613). The BBB score was significantly higher than that in the control group. On the 14th day after operation, the apoptosis rates of control group, MP group, BMSCs group and MP BMSCs group were (48.47 卤5.70)%, (31.95 卤3.58)%, (41.39 卤2.33)% and (23.48 卤2.69)%, respectively. P0.0001 and BMSCs group (QM6.716P0. 0002) were significantly lower than those of control group, MP BMSCs group (QM7.332P0. 0001) and BMSCs group (QU15. 460 P0. 0001). On the 14th day after operation, the results of immunohistochemical staining showed that the MP group (QN 14.710 P0.0001) and the BMSCs group (QV6.502P0. 0001); The number of TNF- 伪 and IL-1 尾 positive cells in MP group was significantly lower than that in control group. The number of IL-1 尾 positive cells in MP group and BMSCs group were significantly lower than those in control group. The number of TNF- 伪 and IL-1 尾 positive cells was significantly higher than that of MP BMSCs group (P 0.0001). The positive rate of Brd U in BMSCs group was (6.600 卤0.3399)%, which was significantly lower than that in MP BMSCs group (9.300 卤0.5175)% (t = 4.361). Conclusion MP can significantly improve the recovery of hindlimb motor function in rats with SCI treated by BMSCs transplantation. Both of them can reduce the expression of TNF- 伪 and IL-1 尾 and MP can effectively enhance the survival of the transplanted BMSCs.
【作者单位】：河北北方学院附属第一医院骨科;河北北方学院附属第一医院研究生院;
【基金】：河北省卫生厅医学科学研究重点项目计划(20110176) 河北北方学院创新人才培育基金项目(CXRC1322)~~
【分类号】：R651.2

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