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目的探讨经鼻给予碱性成纤维细胞生长因子(bFGF)对脑梗死大鼠内源性神经干细胞再生的影响。方法取成年雄性SD大鼠36只,随机分为假手术组(n=12)、对照组(n=12)、bFGF组(n=12)。对照组和bFGF组大鼠采用大脑中动脉线栓法建立局灶性脑缺血-再灌注模型,术后第1~6天,分别经鼻给予等渗盐水(30μl)或bFGF(每天30μg,30μl)。在术前及术后第1、7、14、21、28天对脑梗死后大鼠神经功能恢复的程度进行改良神经功能缺损评分(mNSS)。采用苏木素染色法测定脑梗死大鼠在术后第7、28天的脑梗死体积;采用免疫组化法测定术后第7和28天缺血侧室管膜下区(SVZ)及纹状体5-溴脱氧尿核苷(BrdU)阳性细胞数;激光共聚焦观察第7天时BrdU与doublecortin(DCX)的免疫荧光双标染色情况。结果与对照组相比,bFGF组从术后第7天开始,大鼠神经功能就显著恢复,术后第1、7、14、21、28天,对照组mNSS评分分别为8.7±1.0、7.2±0.8、6.7±1.0、6.2±0.8、5.8±1.2;bFGF组分别为8.8±0.8、6.2±0.8、5.3±0.8、5.0±0.6、4.3±0.8。两组间各时间点相比,P值分别为0.756、0.044、0.033、0.016、0.028,显示了其明显的正性干预作用。而对照组与bFGF组大鼠的脑梗死体积差异无显著性。假手术组大鼠无梗死灶。BrdU免疫组化显示,术后第7和28天,经鼻给予bFGF组大鼠在SVZ和纹状体区域BrdU阳性细胞数与对照组相比均显著增高,第7天时,SVZ的大部分BrdU阳性细胞共标不成熟神经元标记物——DCX。假手术组大鼠无明显神经功能缺损症状。结论经鼻给予bFGF,能够诱导脑梗死大鼠内源性神经干细胞的增殖和存活,促进神经功能的恢复。经鼻腔给予bFGF是一种有前途的治疗脑梗死的方法。
Objective To investigate the effect of nasal administration of basic fibroblast growth factor (bFGF) on the regeneration of endogenous neural stem cells in cerebral infarction rats. Methods Thirty - six adult male Sprague Dawley rats were randomly divided into sham operation group (n = 12), control group (n = 12) and bFGF group (n = 12). In the control group and bFGF group, focal cerebral ischemia-reperfusion model was established by middle cerebral artery occlusion method. The rats were anesthetized by nasal administration of isotonic saline (30μl) or bFGF (30μg / 30 μl). The modified neurological deficit score (mNSS) was used to assess the degree of neurological functional recovery after cerebral infarction in preoperative and postoperative 1st, 7th, 14th, 21st and 28th days. The infarct volume of cerebral infarction rats was detected by hematoxylin-eosin staining on the 7th and 28th day after operation. Immunohistochemistry was used to measure the infarction volume in ischemic subventricular zone (SVZ) and striatum 5 BrdU positive cells were observed by laser scanning confocal microscope. Double immunofluorescence staining of BrdU and doublecortin (DCX) on day 7 was performed. Results Compared with the control group, the neurological function of rats in bFGF group recovered significantly on the 7th day after operation. On the 1st, 7th, 14th, 21st and 28th days after operation, the mNSS scores of the control group were 8.7 ± 1.0 and 7.2 ± 0.8,6.7 ± 1.0,6.2 ± 0.8,5.8 ± 1.2; bFGF group was 8.8 ± 0.8,6.2 ± 0.8,5.3 ± 0.8,5.0 ± 0.6,4.3 ± 0.8. P values were 0.756,0.044,0.033,0.016,0.028 at each time point between the two groups, showing a significant positive intervention. There was no significant difference in cerebral infarction volume between control group and bFGF group. Rats in sham operation group had no infarction. BrdU immunohistochemistry showed that on the 7th and 28th day after operation, the number of BrdU positive cells in the bFGF group was significantly higher than that in the control group in the bFGF group. On the 7th day, the majority of BrdU Positive cells co-labeled immature neuronal markers - DCX. Rats in sham operation group had no obvious neurological deficit symptoms. Conclusion Nasal administration of bFGF can induce the proliferation and survival of endogenous neural stem cells in rats with cerebral infarction and promote the recovery of neurological function. Nasal administration of bFGF is a promising method of treating cerebral infarction.