不同频率提插水沟穴对大鼠局灶性脑缺血再灌注皮质细胞凋亡及NF-κBp65、Caspase-3的影响

Effect of acupuncture on GV26 with lift-thrusting manipulation of different frequencies specifically regulating the expression of cortical cell apoptosis, NF-κB p65 and Caspase-3

关键词:  针刺  穴  水沟  局灶性脑缺血再灌注  刺激参数/针刺效应

Key words:  acupuncture  acupoint  Shuigou  focal cerebral ischemia-reperfusion  stimulus parameter/acupuncture effect

摘要:

[目的] 研究不同频率提插水沟穴对大鼠局灶性脑缺血再灌注皮质缺血区NF-κB p65和Caspase-3的影响,寻求适宜针刺频率。[方法] 将成年Wistar雄性大鼠随机分为空白组、假手术组、模型组、水沟穴低频提插组、水沟穴中频提插组、水沟穴高频提插组,每组大鼠6只。空白组不造模,假手术组造模时在颈内动脉内不插入线栓,其余4组均完成局灶性脑缺血再灌注模型。空白组、假手术组和模型组均常规饲养,不进行针刺干预;水沟穴低频提插组、水沟穴中频提插组、水沟穴高频提插组均进行对应频率的针刺干预。每日两次,针刺3 d.TUNEL法检测各组大鼠皮质细胞凋亡的表达,免疫荧光检测大脑皮质NF-κB p65和Caspase-3的表达。[结果] 皮质细胞凋亡的表达：各针刺组与模型组比较(P<0.01)有统计学差异,针刺低频组和针刺中频组及高频组差异有均统计学意义(P<0.01及P<0.05),针刺中频组和高频组比较无统计学差异(P>0.05)).大脑皮质NF-κB p65和Caspase-3的表达：各针刺组与模型组比较(P<0.01)有统计学差异,针刺低频组和高频组有统计学差异(P<0.01),针刺中频组和高频组比较无统计学差异(P>0.05).[结论] 针刺各个频率组都可显着降低脑缺血再灌注损伤所致的细胞凋亡,其中针刺中频组和高频组要显着优于低频组,其可能与降低皮质细胞NF-κB p65和Caspase-3的表达量有关。

Abstract:

[Objective] To observe effect of different acupuncture frequencies on NF-κB p65 and Caspase-3 of cerebral cortex region in Focal cerebral ischemia-reperfusion rats, so as to explore the appropriate frequency. [Methods] The 36 male Wistar rats were randomized into 6 groups: control group, false surgery group, model group, Shuigou (GV 26) with low frequency lifting-inserting manipulation (GV26-LOW) group, GV 26 with mid frequency lifting-inserting manipulation (GV26-MID) group and GV 26 with high frequency lifting-inserting manipulation (GV26-HIGH) group. The control group was not subjected to modeling, the false surgery group was operated as modeling groups but without procedure of inserting thread embolism into internal carotid artery, comparatively, the other four modeling groups were all completed the focal cerebral ischemia-reperfusion procedures. Rats in control group, false surgery group and model group were fed routinely without acupuncture treatment; while, rats in GV26-LOW group, GV26-MID group, and GV26-HIGH group were all acupunctured on GV26 with corresponding methods twice a day for three days. Expression of cortical cell apoptosis was detected by TUNEL method and expression of NF-κB p65 and Caspase-3 in cerebral cortex by immunohistochemistry. [Results] Expression of cortical cell apoptosis in 3 acupuncture groups were significantly lower than that in model group (P<0.01), moreover, its expression in GV26-MID group and GV26-HIGH group were significantly lower than that in GV26-LOW group (P<0.01 or P<0.05). While, there was no statistical difference in cortical cell apoptosis expression between GV26-HIGH group and GV26-MID group (P>0.05). Expression of cortical NF-κB p65 and Caspase-3 in 3 acupuncture groups were significantly lower than that in model group (P<0.01), moreover, its expression GV26-HIGH group were significantly lower than that in GV26-LOW group (P<0.01). While, there was no statistical difference in cortical NF-κB p65 and Caspase-3 between GV26-HIGH group and GV26-MID group (P>0.05). [Conclusion] Acupuncture with 3 different manipulation frequencies could significantly reduce the apoptosis caused by cerebral ischemia-reperfusion, while acupuncture with mid frequency and high frequency is superior to acupuncture with low frequency in the management of cerebral ischemia-reperfusion damage, which may be regulated by expression of NF-κB p65 and Caspase-3 in cerebral cortex.