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目的:探讨ndrg2基因在大鼠脑缺血再灌注后于侧脑室室管膜下区的表达,并探索其规律。方法:成年雄性Sprague-Dawley大鼠,利用大脑中动脉栓塞法造成短暂性脑缺血(MCAO),缺血后2 h再灌注,制备损伤模型。分为假手术组(n=6)、缺血后4 h(n=8)、12 h(n=8)和24 h(n=8)四组。假手术组除不用线栓阻塞大脑中动脉外,其余与缺血组相同。采用原位杂交和Western blot方法观察,MCAO后不同时间点NDRG2在大鼠脑内的表达特点。结果:原位杂交结果显示:NDRG2 mRNA集中表达于侧脑室室管膜下区,阳性细胞呈蓝紫色表达于细胞质,细胞核未见表达。脑缺血再灌注后NDRG2阳性细胞数随时间延长而增多,各缺血组(4 h、12 h和24 h)与假手术组相比均具有统计学差异(P<0.05),其中缺血24 h组的阳性细胞数最多;Western blot结果与原位杂交结果一致:各缺血组NDRG2蛋白表达水平随着缺血再灌注时间而增加,与假手术组相比具有统计学差异(P<0.05)。结论:NDRG2集中表达于大鼠侧脑室室管膜下区;脑缺血再灌注可以诱导该区内NDRG2表达的明显增高。
OBJECTIVE: To investigate the expression of ndrg2 gene in the subventricular zone of lateral ventricle after cerebral ischemia-reperfusion in rats and explore its regularity. Methods: Adult male Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion (MCAO) by cerebral middle artery embolization method and then reperfused at 2 h after ischemia. The rats were divided into sham operation group (n = 6), 4 h after ischemia (n = 8), 12 h (n = 8) and 24 h (n = 8) In the sham-operated group, except for blocking the middle cerebral artery with a thread plug, the rest were the same as the ischemic group. The expression of NDRG2 in rat brain at different time points after MCAO was observed by in situ hybridization and Western blot. Results: In situ hybridization results showed that NDRG2 mRNA was concentrated in the subependymal zone of the lateral ventricle. The positive cells were expressed in the cytoplasm in the form of blue-purple and no expression in the nucleus. The number of NDRG2 positive cells increased with time after cerebral ischemia-reperfusion, and there was a significant difference (P <0.05) between ischemic group (4 h, 12 h and 24 h) and sham operation group 24 h group had the highest number of positive cells; Western blot results were consistent with in situ hybridization results: NDRG2 protein expression in each ischemic group increased with ischemia-reperfusion time, compared with the sham operation group was statistically significant (P < 0.05). CONCLUSION: NDRG2 is mainly expressed in the subependymal zone of rat lateral ventricle. Cerebral ischemia-reperfusion can induce a marked increase of NDRG2 expression in this area.