针刺对缺血再灌注大鼠脑细胞凋亡的保护作用及机制

目的 探讨脑缺血再灌注后针刺对脑细胞线粒体跨膜电位的保护作用及保护机制.方法 采用线栓法制备大鼠局灶性脑缺血再灌注模型,针刺水沟、百会二穴,通过Rhodamin123荧光染色、TUNEL原位标记及电镜观察,分别于脑缺血再灌注后24h及48h检测脑细胞线粒体跨膜电位、线粒体超微结构及脑凋亡细胞的变化.结果 脑缺血再灌注后Rhodamin123的荧光强度增强,术后24h是(961.27±34.22),48h是(1231.23±121.54),与假手术组(782.82±57.24)比较明显增高(P<0.05);针刺干预后Rhodamin123的荧光强度术后24h是(808.44±56.23),48 h是(954.25±35.11),与脑缺血再灌注组比较明显下降(P<0.05);脑缺血再灌注后24h凋亡细胞是(26.12±2.75)个/视野],48 h凋亡细胞是(41.24±4.73)个/视野],与假手术组(3.56±0.92)个/视野]比较明显增加(P<0.05),针刺干预后24h(19.88±3.32)个/视野]与48 h(33.23±3.81)个/视野]凋亡细胞数量与脑缺血再灌注组比较明显减少(P< 0.05);脑缺血再灌注后线粒体膜肿胀、崩解,内嵴断裂,中毒颗粒增加,针刺干预后线粒体的非特异性改变明显减轻.结论 针刺能够稳定线粒体膜的通透性,减少线粒体跨膜电位的耗散,减少缺血再灌注后脑细胞凋亡,增加脑组织对缺血缺氧的耐受性.

Effects of acupuncture on neuronal apoptosis of rats with cerebral ischemia and reperfusion

Objective To investigate the protective effect and mechanisms of acupuncture on the mem-brane potential in mitochondria of the brain cells injury induced by ischemia and reperfusion. Method The mid-dle cerebral artery occlusion/repeffusion (MCAO/R) rat model was established by the modified Longa occlusion method. The Baihui and Shuigou Point in Du meridian was acupunctured electrically 30 min. After reperfusion 24 h and 48 h, mitochondrial membrane potential, apoptosis and mitochondria ultrastrocture of brain cells were detected by the method of Rhodamin123 fluorescence staining,TUNEL in situ labeling and electron microscopy respectively after repeffusion 24 h and 48 h. Results The fluorescence intensity of Rhodamin123 was 961.27±34.22 and 1231.23±121.54 after focal cerebral ischemic reperfusion 24 h and 48 h respectively,this result was increased to compare with sham-operated group. The fluorescence intensity of Rhodamin 123 in group of using the intervention of acupuncture was 808.44±56.23 and 954.25±35.11 after operation 24 h and 48 h respectively ,this values de-creased significantly to compare with the group of ischemic reperfusion. The amount of apoptosis cells after cerebral ischemia reperfusion 24 h and 48 h were more than the sham group. The amount of apoptosis cells after acupuncture was less than the cerebral ischemic reperfusion group. The mitochondrial membrane shown intumesee,disaggrega-tion, endocrista collapsing and toxic granulation increased after ischemie reperfusion and this non-specificity change of mitochondrion alleviated by using acupuncture. Conclusion Acupuncture stabilized the permeability of mitochon-drial membrane, reduced the dissipation of mitochondrial transmembrane potential, decreased the amount of apopto-sis of brain cells of ischemic reperfusion and increased the toleration of brain against the ischemic and anoxie inju-ry. Conclusion Acupuncture could stabilized the permeability of mitochondria membrane. It could decreased the dissipation of cytochondriome transmembrane potential and the amount of apoptosis of brain cells. It could increased the toleration of brain against the ischemic and anoxic injury.