Chen Huai-long, Qi Hui, Liu Xiao-jie, Wang Ming-shan
Zhen Ci Yan Jiu. 2014 Dec;39(6):431-6.
To observe the effect of electroacupuncture (EA) pretreatment on the number of survival neurons and the expression of glucose regulated protein 78 (GRP 78) and growth arrest and DNA damage-inducible gene 153 (GADD 153) in the hippocampus in rats with global cerebral ischemia /reperfusion injury (CI/R), so as to study its underlying mechanism in neuroprotective action.
SD male rats were randomly divided into 3 groups (n =48 each):sham operation,Cl/R model and EA pretreatment group. Global CI/R model was induced by 4-vessel occlusion (bilateral vertebral artery cauterization and bilateral carotid artery ligation for 5 min, followed by reperfusion). Before modeling, EA pretreatment of "Baihui" (GV 20) and "Dazhui" (GV 14, 2 Hz/15 Hz, 1 mA) was given to rats of the EA pretreatment group for 30 min, once daily for 5 days. At 6, 12, 24 and 48 h after CI/R, the hippocampus tissues of rats in different subgroups were separately sampled to be stained with H. E. method for detecting the number of the survived neurons, stained with TUNEL method for assaying the apoptotic neurons in the CA 1 region, and processed with Western blot (WB) for assaying the expression of GRP 78 and GADD 153 proteins.
Compared with the sham group, the number of hippocampal survival neurons was significantly decreased at the time-points of 12 h, 24 h and 48 h after CI/R in the model group (P<0. 05) and was apparently increased by EA pretreatment at 24 h and 48 h (P<0. 05). The number of the apoptotic neurons in the hippocampal CA 1 region at the time-points of 6 h, 12 h, 24 h and 48 h after CI/R was significantly bigger in the model group than in the sham group (P<0. 05), and was obviously decreased at 12 h, 24 h, 48 h after CI/R in the EA pretreatment group (P<0.05). WB detection showed that the expression levels of hippocampal GRP 78 and GADD 153 proteins at the four time-points after CI/R were significantly higher in the model group than in the sham group (P<0. 05). Compared with the model group, hippocampal GRP 78 protein expression levels at the 4 time-points were further markedly up-regulated (P<0.05), while GADD 153 protein expression levels at the 4 time-points were significantly suppressed in the EA pretreatment group (P<0.05).
EA pretreatment can effectively suppress the number of hippocampal apoptotic neurons and increase survival rate of neurons in CI/R rats, which may be closely associated with its effects in up-regulating the expression of GRP 78 protein and down-regulating the expression of GADD 153 protein in the hippocampus.
观察电针预处理对全脑缺血/再灌注损伤（CI/R）大鼠海马区存活神经元数量及葡萄糖调节蛋白78（GRP 78）、生长停滞和DNA损伤诱导基因153（GADD 153）表达的影响，探讨其神经保护作用的潜在机制。
将雄性SD大鼠随机分为3组（每组n = 48）：假手术组、CI/R模型组和电针预处理组。采用四血管闭塞法（双侧椎动脉烧灼和双侧颈总动脉结扎5分钟，随后再灌注）诱导全脑CI/R模型。在建模前，对电针预处理组大鼠进行“百会”（GV 20）和“大椎”（GV 14，2Hz/15Hz，1mA）电针预处理30分钟，每天1次，共5天。在CI/R后6、12、24和48小时，分别取不同亚组大鼠的海马组织，采用苏木精-伊红（H.E.）染色法检测存活神经元数量，采用TUNEL法检测CA1区凋亡神经元，采用蛋白质免疫印迹法（WB）检测GRP 78和GADD 153蛋白表达。
与假手术组相比，模型组在CI/R后12、24和48小时海马存活神经元数量显著减少（P<0.05），电针预处理在24和48小时可明显增加存活神经元数量（P<0.05）。模型组在CI/R后6、12、24和48小时海马CA1区凋亡神经元数量显著多于假手术组（P<0.05），电针预处理组在CI/R后12、24、48小时凋亡神经元数量明显减少（P<0.05）。WB检测显示，模型组在CI/R后四个时间点海马GRP 78和GADD 153蛋白表达水平显著高于假手术组（P<0.05）。与模型组相比，电针预处理组在四个时间点海马GRP 78蛋白表达水平进一步显著上调（P<0.05），而GADD 153蛋白表达水平在四个时间点均显著下调（P<0.05）。
电针预处理可有效抑制CI/R大鼠海马凋亡神经元数量，提高神经元存活率，这可能与其上调海马GRP 78蛋白表达和下调GADD 153蛋白表达有关。
