失荅剌知丸对脑缺血大鼠再灌注后神经功能缺损及脑组织PI3-K和AKT表达的影响

The Effect of Shi-da-la-zhi-wan on Phosphoinositide 3-kinase and Serine-threonine Kinase Expression after Cerebral Ischemia-reperfusioninjury in Rrats

目的探讨失荅剌知丸对脑缺血大鼠再灌注后神经功能缺损恢复程度以及皮质缺血周围区磷脂酰肌醇-3-激酶(PI3-K)和丝氨酸/苏氨酸蛋白激酶(AKT)表达的影响。方法 169只SD雄性大鼠随机分为假手术组、模型组、金纳多组(阳性药物对照组)和失荅剌知丸组,后3组采用线栓法制备大脑中动脉闭塞(MCAO)模型,缺血2h后再灌注,术后分12h、24h、72h、7d组,共13组,每组13只。采用Zea Longa评分法分别观察神经功能缺损恢复程度;TUNEL法检测24h脑细胞凋亡情况;免疫组化法观察各组大鼠脑内PI3-K和AKT的表达情况。结果与假手术组比较,模型及各药物组神经功能评分均降低(P<0.05);与模型组比较,各用药组24h、72h和7d神经功能恢复明显,神经功能评分上调,以72h和7d点改善尤为明显(P<0.01);失荅剌知丸72h、7d组神经功能评分较金纳多组显著提高(P<0.01)。24h脑细胞凋亡情况显示:失荅剌知丸组在24h组凋亡阳性细胞数明显低于金纳多组和模型组(P<0.01)。与假手术组比较,模型组、各药物组PI3-K和AKT的表达增高(P<0.01);与模型组比较,各药物组PI3-K和AKT的表达增高(P<0.01);与金纳多组比较,失荅剌知丸组PI3-K和AKT的表达增高,其中24h、72h、7d表达明显(P<0.01)。结论失荅剌知丸可明显改善脑缺血/再灌注损伤大鼠的神经功能,抑制神经细胞凋亡;上调PI3-K和AKT的表达,激活PI3-K/AKT信号通路,可能是其发挥神经保护作用的机制之一。

Objective To investigate the effects of shi - da - la - zhi - wan on phosphoinositide3 - kinase （PI3K） and serine- threonine kinase （AKT） expression after cerebral i~,chemia- reperfusioninjury in rats. Methods The ischemia model was established by right middle cerebral artery occlusion （MCAO）, schemia reperfusion after 2h, 169 SD rats were divided into sham operation group, madel group, shi - da - la - zhi - wan group ,jin- na- duo group. Scores were observed using the neural functiotL after surgery for 12h ,24h ,72 and 7d. Apoptosis was detected by TUNEL method. Immunohistochemical metaod was used to observe each group PI3 - K in rats and AKT expression. Results Compared with those of sham group, the model group were significantly lower at each time point on neurological deficit scale scores（ D 〈 0.01 ） ;Compared with those of model group,the shi- da- la- zhi- wan group and jin- na- duo group vrere evidently superior at each time point on neurological deficit scale seores（P 〈0.01 ） ;compared with those of Jin- na- duo group,hui the shi- da- la- zhi -wan group were significantly higher at 72h,7d time point on neurological deficit scale scores（P 〈 0.01 ） ;The number of TUNEL- stained apoptoticcells was also lower in shi -da -la -zhi -wan groups compared to all others at 24 hour（P 〈0.01 ）. Compared with those of Mod- el group, shi - da - la - zhi - wan group and jin - na - duo group was significantly higher on PI3 - K and AKT expression （ P 〈 0.01 ）. Shi - da - la - zhi - wan group and Jin - na - duo group , two group difference is sig- nificant at 24h,72h,7 d （ P 〈 0.01 ）. Conclusion Shi - da - la - zhi - wan can improve the neural functional recovery in rats, inhibit neural cell apoptosis, increase the PI3 - K and AKT and activate the PI3 - K/AKT sig- naling pathway.