SIRT1对大鼠脊髓损伤神经功能恢复的影响

发布时间：2018-11-28 11:51

【摘要】：目的：探索SIRT1与大鼠脊髓损伤后炎症因子，细胞凋亡及神经功能恢复的关系。方法：采用改良Allen s法，建立大鼠脊髓钝挫伤模型（T10），，112只SD大鼠被随机分成4组：假手术组（n=16只），模型组（n=32只），白藜芦醇组（n=32只），对照剂组（n=32只）。脊髓损伤术后立即给予腹腔注射白藜芦醇（resveratrol,RES），100mg/kg和聚乙二醇硬脂酸酯15(solutol HS 15,SOL)，100mg/kg.进行功能评分的大鼠术后每天给药一次，连续1周。分别采用QT-PCR方法、ELISA方法、TUNEL荧光标记法对4组（SHAM，RES，SOL，SCI）脊髓组织中各时间点SIRT1的表达活性、炎症因子变化和细胞凋亡情况进行检测，并对大鼠术前和术后1天，3天，1周，2周，3周，4周进行BBB scores功能评分结果：所有引物都可以成功的扩增出目的片段。RES组，SCI组和SOL组3组SIRT1的表达都有一定升高，RES组SIRT1表达在4h明显增加，在8h达到高峰，与其他两组(SCI组, SOL组)比较在4h,24h,36h差别有统计学意义（P＜0.05）.除假手术组以外各组自8h后SIRT1表达下降。 SCI组大鼠损伤脊髓组织在损伤后4 h各炎症性细胞因子表达即明显升高,其中TNF-α和IL-6于损伤后8 h达峰值,IL-10在观察时间内呈持续性增高，IL-1β在24h达峰值。SOL组各炎症因子的变化规律与SCI组基本一致，RES组与前两组相比，浓度差别均有统计学意义（P＜0.01）。与SHAM组相比，SCI组，RES组，SOL组术后1天凋亡细胞明显升高，统计学比较差异性有意义（p＜0.05）。在3天时，SCI组，RES组，SOL组凋亡细胞数持续升高分别达到54.82，38.49，53.24，与其他两组比较，RES组升高较低差异有统计学意义（p＜0.05），通过定位发现，凋亡主要存在于神经元。术后1天除假手术组功能评分没有明显改变外，其他3组后肢功能完全丧失0分,从术后第三天到第3周呈现逐渐升高趋势。SCI组，RES组，SOL组比较，前3周功能评分差别没有统计学意义（p＞0.05）。在第四周时，RES组功能评分上升比较明显，与SCI组、SOL组比较差别有统计学意义（P＜0.05），可见通过上调SIRT1的表达活性，能够促进大鼠脊髓损伤后功能恢复。结论: SIRT1可能是通过抗炎，抗凋亡作用，促进大鼠脊髓损伤后功能恢复,其具体机制需要进一步实验。
[Abstract]:Aim: to investigate the relationship between SIRT1 and inflammatory factors, apoptosis and neural function recovery after spinal cord injury in rats. Methods: the spinal cord contusion model (T10) was established by modified Allen s method. 112 SD rats were randomly divided into 4 groups: sham operation group (n = 16), model group (n = 32), resveratrol group (n = 32) and control group (n = 32). Immediately after spinal cord injury, intraperitoneal injection of resveratrol (resveratrol,RES), 100mg/kg and polyethylene glycol stearate (15 (solutol HS 15 mol / kg) was performed immediately after spinal cord injury. Rats with functional score were given drugs once a day for 1 week after operation. The expression of SIRT1, the changes of inflammatory factors and apoptosis in spinal cord tissues of four groups (SHAM,RES,SOL,SCI) were detected by QT-PCR method, ELISA method and TUNEL fluorescence labeling method at different time points, respectively. The results of BBB scores functional score of rats before and 1 day, 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks after operation were as follows: all primers could successfully amplify the target fragment. RES group. The expression of SIRT1 in SCI group and SOL group increased to a certain extent, and the expression of SIRT1 in RES group increased significantly at 4 h and reached the peak at 8 h, which was significantly higher than that in the other two groups (SCI group, SOL group) at 4 h, 24 h and 36 h (P < 0 05). The expression of SIRT1 in all groups except sham-operation group decreased after 8 hours. In SCI group, the expression of inflammatory cytokines in injured spinal cord tissue increased significantly at 4 h after injury. The expression of TNF- 伪 and IL-6 reached its peak at 8 h after injury, and IL-10 increased continuously during the observation time. The change of IL-1 尾 in SOL group was basically the same as that in SCI group, and the concentration difference between RES group and the former two groups was statistically significant (P < 0. 01). Compared with SHAM group, SCI group, RES group and SOL group significantly increased the apoptotic cells 1 day after operation (p < 0. 05). On the 3rd day, the number of apoptotic cells in SCI group, RES group and SOL group continued to increase to 54.82%, 38.49% and 53.24% respectively. Compared with the other two groups, the increase of apoptotic cells in RES group was significantly lower than that in other two groups (p < 0. 05). Apoptosis mainly occurs in neurons. On the first day after operation, the functional score of the other three groups was completely lost except that the functional score of the sham operation group did not change significantly, and the function of the hind limbs was gradually increased from the third day to the third week after operation. The comparison among the SCI group, the RES group and the SOL group was made. There was no significant difference in functional score in the first 3 weeks (p > 0.05). At the fourth week, the functional score of RES group was significantly increased, which was significantly higher than that of SCI group and SOL group (P < 0. 05). It can be seen that by upregulating the expression of SIRT1, it can promote the recovery of function after spinal cord injury in rats. Conclusion: SIRT1 may promote the functional recovery after spinal cord injury through anti-inflammatory and anti-apoptotic effects, and its specific mechanism needs further experiments.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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