醒脑静联合丁苯肽对脑缺血再灌注损伤后细胞凋亡的影响

Effects of Xingnaojing injection combined with butylphthalide(NBP)on the expression of apoptosis after cerebral ischemia reperfusion

目的观察醒脑静、丁苯酞及二者联合分别对大鼠脑缺血再灌注损伤后神经细胞凋亡及Bcl-2和Bax表达的影响。方法 60只雄性wistar大鼠(250±20)g采用改良线栓法制作脑缺血再灌注损伤模型(Middle cerebral artery occlusion,MCAO),随机分为4组,即模型组、醒脑静组、丁苯酞组、醒脑静联合丁苯酞(联合用药)组,每组又分为6、24、72h三个亚组;通过原位末端转移酶标记技术(TUNEL)检测神经细胞凋亡情况,采用免疫组化法观察大鼠脑缺血再灌注各个时间点Bcl-2、Bax的表达水平。结果 (1)模型组手术对侧大脑半球偶见凋亡细胞,病灶区可见大量神经细胞凋亡。丁苯酞用药组、醒脑静用药组凋亡细胞数明显减少,醒脑静联合丁苯酞组凋亡细胞数最少(P<0.05);(2)丁苯酞组及联合用药组Bcl-2阳性表达水平较模型组均有提高,联合用药组Bcl-2阳性表达水平在各时间点均最高(P<0.05);丁苯酞组及联合用药组Bax阳性表达水平较模型组均有降低,联合用药组Bax阳性表达水平最低(P<0.05)。结论 (1)醒脑静、丁苯酞及二者联合均可能通过抑制脑缺血再灌注损伤后神经细胞凋亡来实现神经细胞保护作用,其中二者联合效果最佳;(2)丁苯酞可能通过增加脑缺血再灌注损伤大鼠Bcl-2表达,减少Bax表达的方式来减少神经细胞凋亡,从而减轻脑缺血再灌注损伤;(3)醒脑静本身不能对Bcl-2、Bax的表达水平产生影响,但其可能通过增强丁苯酞作用的方式影响Bcl-2、Bax的表达,从而减轻脑缺血再灌注损伤。

Objective To observe the effects of Xingnaojing injection,butylphthalide（NBP）and combined with them on the expression of apoptosis,Bcl-2and Bax in rats after cerebral ischemia reperfusion.To investigate whether there are protective effects on brain,and to explore its possible mechanism.Methods Sixty male Wistar rats by modified suture method for cerebral ischemia reperfusion（MCAO）were randomly assigned into four groups：model group,Xingnaojing group,butylphthalide（NBP）group,combined with NBP and xingnaojing（combination）group.Each group was divided into three subgroups：6h,24 hand 72h.The apoptosis of nerve cells was detected by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling（TUNEL）,and the expression of Bcl-2and Bax were detected by immunohistochemistry.Results In the model group,a large number of apoptotic cells were observed in the lesion area.In NBP group and Xingnaojing group,the number of apoptosis were significantly reduced,the combination group at the least level（P〈0.05）.In NBP group and combination group,the expression of bcl-2 was significantly lower than that in the model group.The combination group at the highest level,at each time point（P〈0.05）.The expression of bax was to the contrary（P〈0.05）.Conclusion The use of xingnaojing,NBP and combination of the two have neuroprotective effects through the inhibition of nerve cell apoptosis,in which the combination group do the best.NBP can relieve the brain damage by up-regulating the expression of Bcl-2and down-regulating the expression of Bax.Xingnaojing injection itself cannot influence the expression of bcl-2and Bax.But it can enhance the effect of NBP to the expression of bcl-2and Bax.It can indirectly alleviate cerebral ischemia reperfusion injury.