目的探讨神经巢蛋白(nestin)和胰岛素样生长因子-1(IGF-1)在大鼠脑缺血再灌注损伤后脑组织内表达变化的关系及中药通心络干预的影响。方法制作大鼠缺血再灌注损伤(MCAO)模型,设大、小剂量组和对照组。大、小剂量组按照每只1g/(kg·d)、0.5g/(kg·d)给予2ml通心络溶液,2/d,分别给药至造模后3、5、7、14、21、30d,对照组同时以等量生理盐水灌胃。应用荧光免疫组化法检测缺血后缺血侧室管膜下区(SVZ)、海马齿状回(DG)区域nestin的表达,RT-PCR法检测缺血病灶周围脑组织内IGF-1mRNA的表达变化。结果经通心络治疗后,各时间点SVZ、DG区nestin阳性细胞荧光强度值均高于对照组(P〈0.01),其中14d时点最高。除外21、30d时点,其他各时点大剂量组nestin阳性细胞的数目均多于小剂量组(P〈0.01)。大、小剂量组缺血病灶周围脑组织内IGF-1 mRNA的含量与对照组相比,除3d时点无差异以外,其余时点均高于对照组(P〈0.01),但通心络大、小剂量组各时点的IGF-1 mRNA含量无显著差异。结论通心络可明显促进大鼠缺血再灌注损伤后SVZ、DG区神经干细胞的增殖,其部分机制可能与其诱导缺血病灶周围脑组织内IGF-1 mRNA的表达增加有关。
Objective To investigate the expression of nestin protein and insulin-like growth factor 1 (IGF-1) mRNA in rat brain injured by ischemia-reperfusion (I/R), and observe the effects of Tongxinluo, one of Chinese herb drugs, on the expression. Methods The rat model of cerebral I/R injury, reproduced by middle cerebral artery occlusion (MCAO) and release, were divided into three groups: high-dose Tongxinluo group (given lg/kg·d of Tongxinluo], low-dose Tongxinluo group (given 0. 5g/kg · d of Tongxinluo) and control group (administration of saline). 2ml of Tongxinluo or saline each time was administered to each rat and 2 times every day up to day 3, 5, 7, 14, 21 and 30 after the injury. The expression of nestin in the subventricular zone (SVZ) and hippoeampal dentate gyrus zone (DG) was detected by immunohistochemistry, and the IGF-1 mRNA expression around isehemic area was detected by RT-PCR. Results Compared with the control group, the immunofluorescence magnitude of nestin-positive neurons in SVZ and DG was significantly higher in high- and low-dose Tongxinluo groups (P〈0.01) at each time point, especially on day 14. The immunofluorescence magnitude of nestin-positive neurons was remarkably higher in high-dose Tongxinluo group than in low-dose Tongxinluo group (P〈0. 01) at all time points except at day 21 and day 30. The expression of IGF-1 mRNA around ischemic area significantly increased at all time points except day 3 in both high- and low-dose Tongxinluo groups (P〈0. 01) compared with that in control group. However, no significant difference existed between the high- and low-dose Tongxinluo groups at each time point (P〉0. 05). Conclusions Tongxinluo, a kind of Chinese herb drugs, may promote the proliferation of neural stem cells in SVZ and DG after cerebral I/R injury in rats, and the mechanism may be related to upregulation of IGF-1 mRNA induced by Tongxinluo.