清脑益元汤对大鼠脑缺血损伤Nestin、NGF表达的影响

目的研究清脑益元汤对大鼠脑缺血损伤后巢蛋白(Nestin)、神经生长因子(NGF)表达的影响,探究清脑益元汤对缺血性脑损伤神经元保护作用的部分机制。方法将192只健康SD大鼠随机分为两大组,每大组再分为4个亚组:空白组(n=6)、假手术组(n=30)、模型组(n=30)、清脑益元汤组(n=30)。采用改良的Longa线栓法制备大鼠急性脑缺血损伤模型,除空白组外,假手术组、模型组及清脑益元组按缺血损伤后1 d、3 d、7 d、14 d、28 d 5个取材时间点分为5个亚组。造模成功后取大鼠缺血侧脑组织,采用免疫组化法检测大鼠缺血侧皮质区Nestin、NGF表达,采用实时荧光定量PCR(Real-Time PCR)法检测大鼠缺血侧皮质区Nestin、NGF mRNA表达。结果(1)免疫组化法:与假手术组相比,模型组、清脑益元组在缺血侧Nestin、NGF阳性细胞明显增多(P<0.01,P<0.01);与模型组相比,清脑益元组Nestin、NGF阳性细胞表达明显增多(P<0.01,P<0.05);(2)Real-Time PCR法:与假手术组相比,模型组、清脑益元汤组大鼠脑组织缺血侧皮质区Nestin、NGF mRNA表达量增加(P<0.01,P<0.01);与模型组相比,清脑益元汤组大鼠脑组织缺血侧皮质区Nestin、NGF mRNA表达量升高(P<0.01,P<0.05)。结论清脑益元汤可能通过上调脑缺血损伤后Nestin、NGF蛋白及mRNA表达,促进脑组织损伤修复、保护神经元,进而发挥脑保护的作用。

Effect of QingnaoYiyuan decoction on expression of Nestin and NGF in ischemic cortex of rats with cerebral ischemic injury

Objective To explore the effect of QingnaoYiyuan decoction on the expression of Nestin and nerve growth factor(NGF)in the ischemic cortex of rats with cerebral ischemic injury and the possible mechanism of neuronal protection.Methods 192 healthy SD rats were randomly divided into two groups.Each group was subdivided into 4 subgroups:blank,sham,model and QingnaoYiyuan decoction groups.The rat model of acute cerebral ischemic injury was established by modified Longa thread occlusion method,and the rats were divided into 5 subgroups according to the time points after operation:1,3,7,14 and 28 days.The expressions of Nestin and NGF in the ischemic cortex were detected by immunohistochemical method,and the expressions of Nestin and NGF mRNA in the ischemic cortex were detected by real-time fluorescence quantitative PCR(Real-time PCR).Results Compared with sham group,the expressions of Nestin and NGF positive cells in the ischemic cortex of model group and QingnaoYiyuan group were significantly increased(P<0.01),and the expressions of Nestin and NGF positive cells in the ischemic cortex of the QingnaoYiyuan group were significantly higher than those of model group(P<0.01).At the same time point,compared with sham group,the expressions of Nestin and NGF mRNA in the ischemic cortex of model group and QingnaoYiyuan decoction group were significantly increased(P<0.01),and the expressions of Nestin and NGF mRNA in the ischemic cortex of QingnaoYiyuan decoction group were significantly higher than those of model group(P<0.01).Conclusions QingnaoYiyuan decoction might play a role in brain protection by up-regulating the expressions of Nestin,NGF protein and mRNA after cerebral ischemia injury,promoting the repair of brain tissue injury and protecting neurons.