摘要
目的观察蛇床子素(Osthole)对大鼠脑缺血再灌注损伤的作用及机制。方法将50只SD大鼠随机分成假手术组、模型组和蛇床子素(25、50、100mg/kg)组(n=10)。利用物理法评估大鼠脑梗死范围、脑含水量和神经症状,试剂盒检测活性氧自由基(ROS)和ATP酶活性,流式细胞仪检测线粒体膜电位,免疫蛋白印迹法检测Caspase3、Clevage-Caspase3、Caspase9、Clevage-Caspase9、Bcl-2、Bax、细胞质中凋亡诱导因子(AIF)和cytochrome C的表达水平。结果与假手术组相比,模型组脑梗死范围、脑含水量、神经症状以及ROS、Cleaved-Caspase3、Cleaved-Caspase9、Bax、细胞质中AIF和cytochrome C的表达明显增加,而Caspase3、Caspase9和Bcl-2表达以及线粒体膜电位和ATP酶的活性明显降低。与模型组相比,蛇床子素组脑梗死范围、脑含水量、神经症状以及ROS、Cleaved-Caspase3、CleavedCaspase9、Bax、细胞质中AIF和cytochrome C的表达明显减少,而Caspase3、Caspase9和Bcl-2表达以及线粒体膜电位和ATP酶的活性明显增加。结论蛇床子素预处理可减轻脑缺血再灌注损伤,其机制可能与抑制线粒体介导的凋亡信号途径相关。
Objective To observe the influence of Osthole on cerebral ischemia-reperfusion injury and its acting mechanism.Methods We randomly divided 50 SD rats into sham(Sham)group,ischemia-reperfusion injury(IRI)group and Osthole(25,50,and 100 mg/kg)(Osthole)groups(n=10).The neurological symptoms,extent of cerebral infarction and cerebral water content were evaluated by physical approach.The activities of ROS and ATP were examined by the kit;the mitochondrial membrane potential(MMP)was measured by flow cytometry.The expressions of Bax,Bcl-2,Caspase3,clevage-Caspase3,Caspase9,clevage-Caspase9,cytoplasmic AIF and cytochrome C were measured by Western blotting.Results Compared with those in sham group,the extent of cerebral infarction and cerebral water content,changes of neurological symptoms,activities of ROS,and expressions of clevage-Caspase3,clevage-Caspase9,Bax,cytoplasmic AIF and cytochrome C in IRI group were significantly increased(P<0.05).However,the activities of ATP,expressions of Caspase3,Caspase9,Bcl-2 and MMP,and ATP activity were decreased(P<0.05).Compared with those in IRI group,the extent of cerebral infarction and cerebral water content,changes of neurological symptoms,activities of ROS,and expressions of clevage-Caspase3,clevage-Caspase9,Bax,cytoplasmic AIF and cytochrome C in Osthole group were significantly decreased(P<0.05).But the activities of ATP,expressions of Caspase3,Caspase9,Bcl-2 and MMP,and ATP activities were increased(P <0.05).Conclusion Osthole pretreatment can attenuate cerebral ischemiareperfusion injury,which is associated with suppressing mitochondria-mediated apoptosis.
引文
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