【摘要】：目的探討甲強(qiáng)龍(MP)在骨髓間充質(zhì)干細(xì)胞(BMSCs)治療大鼠脊髓損傷(SCI)中的作用及對局部腫瘤壞死因子(TNF)-α和白細(xì)胞介素(IL)-1β表達(dá)變化的影響。方法采用改良Allen法制備40只T10脊髓損傷雄性大鼠模型,隨機(jī)數(shù)字表法隨機(jī)分為對照組、MP組、BMSCs組和MP+BMSCs組4組,每組10只。按上述分組分別在脊髓損傷后立即尾靜脈緩慢推注30 mg/kg MP和/或在脊髓損傷后2 h于脊髓損傷部位注射全貼壁法培養(yǎng)并以Brd U標(biāo)記的BMSCs。術(shù)后1、7、14 d,采用BBB運動評分法評價大鼠后肢運動功能恢復(fù)。14 d后處死各組大鼠并取出脊髓樣本,分別進(jìn)行TNF-α、IL-1β免疫組織化學(xué)染色和Tunel染色,各組統(tǒng)計視野內(nèi)TNF-α、IL-1β表達(dá)陽性細(xì)胞數(shù)并檢測細(xì)胞凋亡率;采用Brd U單染觀察BMSCs組和MP+BMSCs組中Brd U陽性率。結(jié)果術(shù)后1 d,各組BBB評分差異無統(tǒng)計學(xué)意義(χ2=1.0756,P=0.7829)。術(shù)后7 d,各組大鼠雙后肢運動功能較前明顯恢復(fù),其中MP組(χ2=8.3265,P=0.0325)和BMSCs組(χ2=14.1166,P=0.0036)的BBB評分明顯高于對照組;MP+BMSCs組的BBB評分明顯高于MP組(χ2=17.7186,P=0.0002)和BMSCs組(χ2=15.8110,P=0.0024)。術(shù)后14 d,各組大鼠雙后肢運動功能明顯恢復(fù),MP+BMSCs組BBB評分明顯高于MP組(χ2=24.7259,P0.0001)和BMSCs組(χ2=25.6014,P0.0001),MP組(χ2=13.5060,P=0.0062)和BMSCs組(χ2=8.9613,P=0.0299)的BBB評分明顯高于對照組。術(shù)后14 d,對照組、MP組、BMSCs組和MP+BMSCs組的細(xì)胞凋亡率分別為(48.47±5.70)%、(31.95±3.58)%、(41.39±2.33)%和(23.48±2.69)%,其中,MP組(q=14.84,P0.0001)和BMSCs組(q=6.716,P=0.0002)明顯低于對照組,MP+BMSCs組明顯低于MP組(q=7.332,P=0.0001)和BMSCs組(q=15.460,P0.0001)。術(shù)后14 d,免疫組織化學(xué)染色結(jié)果顯示,MP組(q=14.710,P0.0001;q=6.710,P0.0001)和BMSCs組(q=6.502,P=0.0001;q=2.849,P=0.0514)的TNF-α及IL-1β陽性細(xì)胞數(shù)均明顯低于對照組,MP組(q=5.573,P=0.0004;q=4.596,P=0.0025)和BMSCs組(q=13.780,P0.0001;q=8.456,P0.0001)的TNF-α及IL-1β陽性細(xì)胞數(shù)明顯高于MP+BMSCs組。術(shù)后14 d,Brd U染色結(jié)果顯示,BMSCs組的Brd U陽性率為(6.600±0.3399)%,明顯低于MP+BMSCs組的(9.300±0.5175)%(t=4.361,P=0.0004)。結(jié)論 MP可明顯提高BMSCs移植治療SCI的大鼠后肢運動功能恢復(fù),二者可協(xié)同降低局部TNF-α和IL-1β表達(dá)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) 偽 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- 偽, IL-1 尾 immunohistochemical staining and Tunel staining were performed respectively. The number of TNF- 偽 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 鹵5.70)%, (31.95 鹵3.58)%, (41.39 鹵2.33)% and (23.48 鹵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- 偽 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- 偽 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 鹵0.3399)%, which was significantly lower than that in MP BMSCs group (9.300 鹵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- 偽 and IL-1 尾 and MP can effectively enhance the survival of the transplanted BMSCs.
【作者單位】： 河北北方學(xué)院附屬第一醫(yī)院骨科;河北北方學(xué)院附屬第一醫(yī)院研究生院;
【基金】：河北省衛(wèi)生廳醫(yī)學(xué)科學(xué)研究重點項目計劃(20110176) 河北北方學(xué)院創(chuàng)新人才培育基金項目(CXRC1322)~~
【分類號】：R651.2

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