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肢体缺血后处理对脑缺血大鼠神经功能恢复和神经细胞增殖的作用

Effects of Limb Ischemic Postconditioning on Behavioral Improvement and Proliferation of Neural Cells in

  • 摘要: 目的探索非侵入式肢体缺血后处理(NLIP)对脑缺血大鼠的脑保护作用和损伤侧皮质区神经细胞的表达与分布。方法SD大鼠110只随机分成假手术组(n=10), 缺血再灌注损伤组(I/R组,n=50),NLIP组(治疗组,n=50)。I/R组和治疗组动物采用线栓法制备大脑中动脉栓塞(MCAO)模型(假手术组不插入线栓),治疗组于再灌注即刻给予NLIP处理(改良的缺血弹性带股动脉近端加压阻断股动脉10 min,再通10 min,循环3次),术后3 d、7 d、14 d、21 d、28 d分别称取大鼠体质量,并进行行为学评估后处死大鼠取脑组织,免疫组织化学染色检测各时点缺血脑皮质中的双皮质素(DCX)和胶质细胞原纤维酸性蛋白(GFAP)的表达,Western blot检测缺血脑皮质中微管相关蛋白2(MAP2)的表达。结果假手术组动物术后1 ~3 d表现出轻度的体质量下降和行为学缺损,术后1 d出现上肢肌力减弱,之后开始恢复。与假手术组比较,以上神经缺损在I/R组和治疗组动物中更加明显(P<0.05),术后3 d到21 d逐渐缓解,到术后28 d时接近假手术组水平。与I/R组相比,治疗组动物体质量在术后2~7 d有明显恢复(P<0.05),动物上肢肌力在14~28 d有明显改善 (P<0.05),治疗组行为学评分在各时点与I/R组差异无统计学意义。与假手术组比较,I/R组和治疗组脑皮质缺血区周围DCX阳性细胞和GFAP阳性细胞随时间推移逐渐增多,伴随胞体肥大和突起变长加粗,分别在术后21 d和28 d大量增殖并聚集在脑皮质缺血区周围。I/R组和治疗组在术后3 d缺血脑皮质中MAP2表达量下降,但与假手术组差异无统计学意义(P>0.05),之后逐渐上调,术后14 d和21 d时高于假手术组(P<0.05)。治疗组MAP2表达量在术后7~28 d高于假手术组(P<0.05),在术后7 d时高于I/R组(P<0.05)。结论NLIP可减轻脑缺血大鼠神经行为学异常和促进缺血周围区神经细胞的增多。

     

    Abstract: Objective To determine the effects of non-invasive limb ischemic postconditioning (NLIP) on the behavioral performance and the expression and distribution of neural cells in rats with ischemic cortex. Methods Rats were randomly divided into 3 groups: sham group (n=10), ischemia/reperfusion (I/R) group (n=50), and NLIP group (n=50). Focal cerebral ischemia was induced by intraluminal MCAO in the rats in the I/R and NLIP groups, while no suture was inserted in the sham-operated rats. NLIP(reperfusion-10 min,ischemia-10 min,×3 cycles) was conducted immediately after reperfusion on bilateral femoral arteries by modified ischemicelastic bands. Body mass and behavioral performance of the rats were assessed at 3 d, 7 d, 14 d, 21 d and 28 d post-ischemia (n=5-10). Doublecortin (DCX) and glial fibrillary acidic protein (GFAP) in the ischemic cortex were detected by immunohistochemistry.The expression of microtubule associated protein 2 (MAP2) was detected by Western blot. Results The rats in the sham group showed mild body mass loss and minor neurological deficit 1-3 d post-ischemia and weakened muscle strength 1 d post-ischemia, which were alleviated gradually overtime. Compared with the sham group, neurological deficits were more obvious in the rats in the I/R and NLIP groups, which were alleviated 3-21 d post-ischemia and reached a level close to that of the sham group at 28 d post-ischemia. No significant difference was found betweenthe I/R and NLIP groups in neurologic deficit scores.NLIP significantly alleviated body mass loss 2-7 d post-ischemia (P<0.05) and improved muscle strength 14-28 d post-ischemia (P<0.05).

     

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