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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3d、7d、14d、21d、28d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1~3d表现出轻度的体质量下降和行为学缺损,术后1d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3d到21d逐渐缓解,到术后28d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2~7d有明显恢复(P<0.05),动物上肢肌力在14~28d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21d和28d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3d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14d和21d时高于假手术组(P<0.05)。治疗组MAP2表达量在术后7~28d高于假手术组(P<0.05),在术后7d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。
Objective To explore the neuroprotective effects of non-invasive post-ischemic (NLIP) on cerebral ischemia in rats and the expression and distribution of neurons in injured cortex. Methods One hundred and ten SD rats were randomly divided into sham operation group (n = 10), ischemia / reperfusion injury group (n = 50) and NLIP group (n = 50). The middle cerebral artery occlusion (MCAO) model was established by thread occlusion in the I / R group and the treated group (the sham operation group was not inserted with thread plug). The treatment group was given NLIP treatment immediately after reperfusion (improved ischemic elastic band femoral artery End compression blocking the femoral artery 10 min, and then through the 10 min, 3 cycles), the rats were weighed at 3d, 7d, 14d, 21d, 28d after operation and subjected to behavioral assessment. (DCX) and glial fibrillary acidic protein (GFAP) expression in ischemic cerebral cortex were detected by immunohistochemical staining. Western blot was used to detect the expression of microtubule-associated protein 2 ( MAP2) expression. Results The animals in sham operation group showed slight weight loss and behavioral defect 1 ~ 3d after operation. The muscle strength of upper limb decreased on the 1st day after operation, and then began to recover. Compared with the sham operation group, the above nerve defects were more obvious in the I / R group and the treated group (P <0.05), gradually relieved from the 3d to 21d after operation, and close to the sham operation group 28d after the operation. Compared with the I / R group, the body weight of the treated group recovered obviously from 2 to 7 days after operation (P <0.05), and the muscle strength of the upper limbs of the treated group was significantly improved from 14 to 28 days (P <0.05) There was no significant difference with I / R group at all time points. Compared with the sham operation group, DCX positive cells and GFAP positive cells in the cerebral cortex ischemic area of I / R group and treatment group gradually increased with the passage of time, accompanied with the enlargement of cytoplasm hypertrophy and protrusion, Proliferate and accumulate around the ischemic area of the cerebral cortex. The expression of MAP2 in ischemic cortex of I / R group and treatment group decreased 3 days after operation, but there was no significant difference with sham operation group (P> 0.05), and then gradually increased after 14 days and 21 days Surgery group (P <0.05). The MAP2 expression in the treatment group was higher than that in the sham operation group (P <0.05) at 7-28 days and higher than that in the I / R group at 7 days after operation (P <0.05). Conclusion NLIP can alleviate the neurobehavioral abnormalities in rats with cerebral ischemia and increase the number of nerve cells in the peri-ischemic area.