灯盏花素对缺血再灌注小鼠脑细胞凋亡的保护作用

 目的探讨灯盏花素对脑缺血再灌注诱导细胞凋亡损伤的保护机制。方法小鼠采用双侧颈总动脉结扎法建立重复全脑缺血再灌注模型。术后24或48h,制备石蜡切片(HE染色)检测脑组织海马CA1区锥体细胞形态学变化;用紫外分光光度计检测脑组织一氧化氮合成酶(NOS)活性;用琼脂糖凝胶电泳技术检测DNA损伤情况,同时用RT-PCR技术检测缺血再灌注过程中小鼠海马CA1区凋亡蛋白抑制剂XIAP(X-chromosome linked inhibitor of apoptosis protein)mRNA的表达。结果全脑重复缺血再灌注可以使海马CA1区幸存的锥体细胞数量减少(P<0.01),提高NOS活性,并增加DNA片段化程度,同时伴随着XIAP mRNA表达显著下调(P<0.01)。5,20和80mg·kg^-1灯盏花素能剂量依赖性减少缺血小鼠海马CA1区神经细胞的死亡(P<0.05,P<0.01),降低NOS(P<0.01)活性,并抑制DNA片段化的产生以及XIAP mRNA表达的下调(P<0.05,P<0.01)。结论灯盏花素可通过抑制神经细胞凋亡而减轻缺血再灌注引起的脑损伤。

Protective Effects of Breviscapilie on Cell Apoptosis Induced by Cerebral Ischemia-Reperfusion in Mice

OBJECTIVE To study the protective effects of breviscapine on cell apoptosis induced by cerebral ischemia-reperfusion in mice.METHODS Repeated cerebral ischemia-reperfusion was induced by fastening bilateral common carotid artery in mice.After 24 or 48 h of reperfusion,the changes of nerve cell morphology of hippocampus CAl area were measured in paraffin section(HE staining).The activity of NOS was then detected by UV-photospectrometer analysis system.Futhermore,DNA Ladder was tested using agarose gel electrophoresis techniques,and the expression of X-chromosome linked inhibitor of apoptosis protein(XIAP) mRNA in hippocampus of repeated ischemia-reperfusion mice was then measured using RT-PCR techniques.RESULTS Repeated ischemia-reperfusion promoted the nerve cell putrescence in hippocampus CAl area(P<0.01),increased the activity of NOS and the degree of DNA segment,and down-regulated the expression of XIAP mRNA(P<0.01).However,breviscapine(5,20,80 mg·kg^-1) dose-dependently reversed the changes induced by cerebral ischemia,remarkably reduced the number of nerve cell putrescence(P<0.05,P<0.01),decreased the activity of NOS(P<0.01),restrained the segment of DNA,and down-regulated the expression of XIAP mRNA(P<0.05,P<0.01).CONCLUSION Breviscapine showed an improvement effect against brain impairment induced by ischemia-reperfusion in mice,and the inhibition of apoptosis in brain may be involved in.