摘要
目的:探讨电针曲池、足三里穴对脑缺血再灌注损伤大鼠缺血周边区皮质、室管膜下区(SVZ)内源性神经干细胞增殖的影响及可能机制。方法:采用改良Zea Longa线栓法制备大鼠脑缺血再灌注损伤模型。电针患肢曲池、足三里穴,治疗3 d和7 d后,观察大鼠神经功能缺损和脑梗死体积情况,以及动物整体行为学的变化;采用免疫组化和免疫荧光法观察Nestin、Vimentin阳性细胞分布及与Brdu共标情况;采用Western Blotting法检测Notch1信号转导通路相关蛋白的表达水平。结果:(1)电针曲池、足三里穴后,脑缺血再灌注损伤大鼠的神经缺损评分降低,脑梗死体积减小(P<0.05);(2)Catwalk步态分析显示,电针治疗3 d和7 d后大鼠通过Catwalk通道的平均速度增加,而持续时间减少(P<0.05);(3)模型组大鼠脑缺血再灌注损伤8d,缺血周围区皮质以及缺血侧SVZ区Ki-67、Nestin和Vimentin阳性细胞数量较缺血再灌注损伤4 d时增加(P<0.05或P<0.01)。电针治疗3 d和7 d均促进神经干细胞增殖(P<0.05);(4)电针治疗7 d后缺血周围区皮质以及缺血侧SVZ区JAG1、NICD、Hes1、Hes5的表达量较模型组明显增加(P<0.05或P<0.01),而电针+GSI组大鼠脑缺血周围区皮质以及缺血侧SVZ区NICD、Hes1、Hes5蛋白的表达量显著降低(P<0.05),但JAG1的表达无统计学意义(P>0.05)。(5)电针+GSI组Nestin阳性细胞、Brdu与Nestin共定位细胞在缺血周围区皮质、缺血侧SVZ区均明显增加(P<0.01或P<0.05)。结论:电针曲池、足三里穴能够减少脑梗死体积,改善动物整体行为学,发挥积极的神经保护作用。其作用机制可能为活化Notch1信号转导通路,释放NICD调控Hes1、Hes5的表达,促进缺血周围区皮质以及缺血侧SVZ区内源性神经干细胞的增殖。
Objective :To explore the effects and possible mechanisms of electroacupuncture(EA) at the Quchi(LI11) and Zusanli(ST36) acupints on the proliferation of endogenous neural stem cells in ischemic surrounding cortex and subventricular zone(SVZ) in ischemia-reperfusion injury rats,and then provide experimental basis for clinical application. Methods:Cerebral ischemia and reperfusion injury model was established by modified Zea Longa suture embolism. EA at Quchi and Zusanli of the affected limb for three days and seven days,the neurological deficit,infarct volume as well as the overall behavior of the animals were evaluated.The distribution of Nestin,Vimentin positive cells and co-labeled with Brdu were observed by immunohistochemistry and immunofluorescence. The expressions of proteins associated with Notch1 signaling pathway were detected by Western Blotting. Results:(1)The neurological deficit scores and infarct volume were reduced after EA at the Quchi and Zusanli acupints in ischemia-reperfusion injury rats(P<0.05).(2) Catwalk gait analysis showed that at the 3rd and 7th day after acupuncture therapy,the average speed increased and the duration decreased when the rats went through Catwalk channel(P<0.05).(3) The numbers of Ki-67,Nestin and Vimentin positive cells in ischemic surrounding cortex and ischemic SVZ region were increased after ischemia-reperfusion injury at8 th day in comparation with 4th day in the model group rats(P<0.05 or P<0.01). The findings of EA treatment for three days and seven days showed that EA could enhance proliferation of neural stem cells(P<0.05).(4) The expression of JAG1,NICD,Hes1,Hes5 in ischemic surrounding cortex and ischemic SVZ area were further increased after EA treatment for seven days in comparation with those in the model group(P<0.05 or P<0.01);and in the EA+GSI group,the expression of NICD,Hes1,Hes5 proteins were significantly decreased(P<0.05),but the expression of JAG1 had no significant difference(P>0.05).(5) In the EA+GSI group,the number of Nestin positive cells and Brdu co-localized with Nestin in ischemic SVZ region were increased(P<0.01 or P<0.05).Conclusions:EA at the Quchi and Zusanli acupints can reduce the infarct volume,improve the overall animal behavior and exert a neuroprotective effect in cerebral ischemia-reperfusion injured rats. The possible mechanism may be activation of the Notch1 signaling pathway resulted in the release of NICD which regulate the expression of Hes1 and Hes5,and thus promote the proliferation of ischemic surrounding cortex area and SVZ-derived neural stem cells.
出处
《康复学报》
2015年第3期23-34,共12页
Rehabilitation Medicine
基金
国家自然科学基金项目(81273835
81373778)
关键词
电针
脑缺血再灌注损伤
神经干细胞
缺血周围区
SVZ区
Notch1信号通路
ectroacupuncture
cerebral ischemia-reperfusion injury
neural stem cells
ischemic surrounding area
SVZ region
Notch1 signaling pathway
