清热化瘀方对脑缺血再灌注损伤大鼠神经干细胞增殖的影响

Influence of Qingre Huayu Prescription on Neural Stem Cell Proliferation in Rats with Cerebral Ischemia/reperfusion

目的观察清热化瘀方对大鼠脑缺血再灌注损伤后神经干细胞增殖的影响。方法 90只大鼠随机分为假手术组、模型组、清热化瘀组3组,采用线栓法制作局灶性脑缺血大鼠(MCAO)模型,观察大鼠缺血损伤后缺血半暗带区的细胞增殖。采用免疫组化法观察脑缺血再灌注后2 d、4 d、7 d、14 d、21 d五个时间点巢蛋白(Nestin)及5-溴脱氧尿苷(BrdU)免疫活性的变化。结果清热化瘀组大鼠在第2天、第4天、第7天的神经功能缺损评分明显优于模型组(P<0.05);模型组缺血半暗带Nestin和BrdU阳性表达分别于再灌注7 d和14 d达高峰(P<0.05),以后表达逐渐减弱;清热化瘀组可以明显增加缺血再灌注后第4天、第7天、第14天大鼠的Nestin阳性细胞表达(P<0.05);并可以明显增加缺血再灌注后第7天、第14天、第21天大鼠缺血半暗带的BrdU阳性细胞表达(P<0.05)。结论脑缺血损伤可激活神经干细胞,促使其增殖和表达。清热化瘀方可以促进MCAO大鼠缺血半暗带神经干细胞的增殖。

Objective To observe the effect of Qingre Huayu prescription（QHP） on proliferation of neural stem cells in rats with cerebral ischemia/reperfusion（I/R）.Methods Ninety rats were randomly divided into three groups：Sham operation group,middle cerebral artery occlusion（MCAO） group and QHP group.The I/R rat models were established by MCAO to observe the expression of nestin and 5 Bromo 2 Deoxy Uridine（BrdU） protein by immunochemistry method at 2 d,4 d,7 d,14 d,21 d after I/R.Results Compared with the model group,the neuro-function deficit score in QHP group were obviously better at 2 d,4 d,7 d（P〈0.05）.The expression of nestin and BrdU protein in the penumbra region were increased at 2 d after reperfusion,and reached peak at 7 d and 14 d separately（P〈0.05）,then decreased continuously.QHP treatment could increase the expressions of nestin at 4 d,7 d,and 14 d,and BrdU at 7 d,14 d,and 21 d（P〈0.05）.Conclusion Cerebral ischemia could stimulate neural stem cells proliferation.QHP could stimulate the proliferation of neural stem cells in rats with MCAO.