摘要
目的在短暂性大脑中动脉栓塞缺血再灌注(transient middle cerebral artery occlusion infarction/reperfusion,tMCAO I/R)模型大鼠中证实电针结合安脑丸治疗对其具有神经保护作用,并对脑梗死后神经保护的机制进行初步探索。方法 SD大鼠随机分为假手术组、模型组、安脑丸组和电针结合安脑丸组。线栓法制作SD大鼠左侧短暂性大脑中动脉栓塞模型。mNSS评分法对各组大鼠各个时间点进行行为学评分。实时荧光定量PCR技术对各组大鼠各个时间点Bcl-2mRNA转录水平进行分析。免疫组化法观察脑梗死后10d各组大鼠脑组织Caspase-3阳性细胞表达,Westernblot分析各组大鼠脑组织Caspase-3蛋白表达水平。TUNEL原位细胞凋亡检测脑梗死后10d各组大鼠梗死区边缘神经元细胞凋亡数量。结果假手术组(正常)mNSS评分为0,未见脑梗死,偶见生理性凋亡细胞。安脑丸组和电针结合安脑丸组在4、7、10dmNSS评分明显低于模型组(均P<0.05),其中,电针结合安脑丸组更低(P<0.05)。实时荧光定量PCR的检测结果显示脑梗死后Bcl-2mRNA转录水平升高,安脑丸组和电针结合安脑丸组与模型组相比升高更明显(均P<0.05),其中,电针结合安脑丸组较安脑丸组更高(P<0.05)。免疫组化及Western blot的检测结果都显示,脑梗死后10d安脑丸组和电针结合安脑丸组大鼠脑组织Caspase-3阳性细胞表达数及蛋白表达水平要明显低于模型组(均P<0.05),其中电针结合安脑丸组较安脑丸组更低(P<0.05)。TUNEL检测结果显示梗死后10d安脑丸组和电针结合安脑丸组的神经元细胞凋亡数量明显低于模型组(均P<0.05),电针结合安脑丸组较安脑丸组更低(P<0.05)。结论中药安脑丸治疗tMCAO I/R模型大鼠,可减少大鼠脑梗死区边缘神经元细胞凋亡数量,改善神经功能,结合电针治疗效果更为显著。其神经保护作用机制可能与针药结合治疗后上调了Bcl-2基因表达,抑制Caspase-3的表达有关。
Objective To examine the neuroprotective effect of the electroacupuncture combined with annao pills on rat cerebral infarction ond the underlying mechanism. Methods SD rats were randomly divided into sham group,model group, annao pill group and electroacupuncture plus annao pill group. The SD rat model of transient left middle cerebral artery occlusion was established by using the suture method. The mNSS scoring was used to assess the behaviors of rats in each group at different time points. Real-time quantitative PCR was performed to detect the Bcl-2 mRNA expression level. Immunohistochemistry was conducted to measure the expression patterns of Caspase-3 in cells of the rat brain tissues. Western blot analysis was used to determine Caspase-3 protein expression levels in each group. Cell apoptosis around the infarcted area in each group was detected by using TUNEL in situ. Results The sham group had the mNSS score of 0,and no cerebral infarction except few physiological apoptotic cells was found. The mNSS score was significantly lower from annao pill group and electroacupuncture plus annao pill group than in model group at 4,7 or 10 days （P〈0.05）. Real-time quantitative PCR showed that the mRNA level of Bcl-2 was significantly increased in rat models of cerebral infaretion, especially in rats from eleetroacupuneture plus annao pill group and annao pill group （P〈0.05）. Immunohistochemistry and Western blot revealed that Caspase-3 positive cells and Caspase-3 protein expression level in electroacupuncture plus annao pill group and annao pill group were much lower than in the model group （P〈0.05）. TUNEL assay results showed that at 10th day after infarction, the number of apoptotic cells around the infarcted area was markedly decreased in both electroacupuneture plus annao pill and annao pill groups as compared with model group （P〈0.05）. The indices of electroacupuncture plus annao pill groups were significantly improved as compared with those of annao pill group （P〈0.05）. Conclusion Annao pill administered after cerebral infarction can improve neurological function and the treatment effect is most manifest when combined with electroacupuncture. The mechanism of the neuroproteetive effect may in- volve the up-regulation of Bcl-2 gene expression, the down-regulation of caspase-3 expression and the decrease in cell apoptosis.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2013年第1期16-21,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30660010)
关键词
安脑丸
电针
脑梗死
神经保护
annao pill
electroacupuncture
cerebral infarction
neuroprotection
