补阳还五汤四类有效部位对大鼠脑缺血再灌注后Caspase表达的作用

The Effct and Mechanism of Buyang Huanwu Decotion(BYHWD) by Caspase Express After Cerebral Ischemia Reperfusion in Rats

目的:研究大鼠局灶性脑缺血再灌注后脑组织半胱氨酰天冬氨酸特异性蛋白酶(Caspase-1、Caspase-3)蛋白表达的变化,探讨补阳还五汤四类有效部位对它的影响。方法:采用线栓法,建立大鼠大脑中动脉栓塞(MCAO)后脑缺血再灌注模型。将SD大鼠随机分为假手术组、模型组、生物碱组、苷组、苷元组、多糖组和尼莫地平对照组,采用免疫组化法测定脑组织Caspase-1、Caspase-3蛋白表达的变化。结果:脑缺血2 h再灌注46 h后,在脉络膜可以见到Caspase-1表达增强,生物碱、苷、多糖、苷元可以抑制Caspase-1表达;在海马区、皮质区和髓质区均可见到Caspase-3蛋白表达增强,生物碱、苷、苷元和尼莫地平可以抑制Caspase-3表达。结论:补阳还五汤具有抗大鼠脑缺血再灌注损伤的作用,生物碱、苷、多糖和苷元可能为其抗缺血性脑损伤的主要物质基础,其作用机制可能与抑制Caspase-1、Caspase-3蛋白的表达,拮抗脑缺血后神经元凋亡有关。

Objcetive： To investigate the effct of the four kinds of active BuYangHuanWu Decotion（BYHWD） by caspase express after cerebral ischemia reperfusion in rats.Further to expound the material base of the BYHWD antagonising cerebral ischemia and the mechanism of BYHWD inhibiting neuronic apoptosis after cerebral ischemia.Methods： Adopt the rat middle cerebral artery occlusion（MCAO） method to induce cerebral ischemia reperfusion model.Rats of SD are randomly divided into 7 groups：sham operation group, model group, alkaloid group（104.54 mg/kg）, glycoside group（168.5 mg/kg）,polysaccharide group（405 mg/kg）,aglycone group（16.5 mg/kg）, and nimodiping group（1 mg/kg）. Polysaccharide group and aglycone group were given medicine in lhour before ischemia, other groups were given medicine in 5 minutes before ischemia, sham operation group and model group were given equal volume normal saline. The express of caspase-1,3 were observed by immunochemistry after cerebral ischemia for 2 hours followed by reperfusion for 46 hours.Resuhs：1. After cerebral ischemia for 2 hours followed by reperfusion for 46 hours, caspase-1 positive reaction increased obviously in the choroid. Aglyeone,glycoside,polysaecharide and alkaloid could inhibit caspase-1 protein express,but nimodiping couldn＇t inhibit caspase-1 protein express.2. After cerebral ischemia for 2 hours followed by reperfusion for 46 hours, caspase-3 positive reaction increased obviously in the hippoeampi,cortex and medulla. Aglycone, glycoside, alkaloid and nimodiping could inhibit caspase-3 protein express,but polysaccharide couldn＇t inhibit easpase-1 protein express.Conclusion： The four kinds of active fractions of BuYangHuanWu Decotion（BYHWD） can antagonize cerebral ischemia injury. Aglyeone,glycoside,polysaccharide and alkaloid can inhibit caspase-1 protein express. Aglycone, glycoside, alkaloid and nimodiping can inhibit caspase-3 protein express.The research revealed aglycone,glycoside,polysaccharide and alkaloid can be the important material base of BYHWD antagonising cerebral ischemia.The mechanism of BYHWD can be associated with inhibiting neuronic apoptosis after cerebral ischemia.