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中文摘要: 目的：观察补益精血方“加减薯蓣丸”对脑缺血大鼠海马神经元及磷酸化丝氨酸/苏氨酸蛋白激酶 (p-serine-threonine kinase,p-Akt)的影响。 方法：40只Wistar大鼠随机分为正常组、假手术组、模型组和中药组,其中模型组、中药组采用改良双侧颈总动脉永久阻断法制作脑缺血大鼠模型,假手术组不予结扎颈总动脉。中药组予加减薯蓣丸10 g·kg-1·d-1灌胃,其余3组用等量0.9%NaCl溶液灌胃。灌胃6周后取材,HE染色观察海马组织形态学变化,免疫荧光法检测神经元核心抗原(NeuN)表达,Western blot法检测p-Akt表达。 结果：HE染色结果显示,模型组大鼠海马细胞密度减少,锥体细胞层变薄,细胞间隙大,排列疏松,细胞核萎缩；中药组大鼠海马细胞密度增加,锥体细胞层变厚,细胞间隙减小,排列紧密,细胞核萎缩减少。模型组大鼠海马组织p-Akt表达较假手术组明显下降(P<0.05),NenN表达较假手术组明显增加(P<0.05)；与模型组比较,中药组海马组织NenN及p-Akt表达均明显增加(P<0.05)。 结论：加减薯蓣丸可促进脑缺血大鼠海马组织p-Akt及NeuN表达,从而可能促进脑缺血引起的大鼠海马神经损伤修复,抑制神经元凋亡。
Abstract: Objective:To observe the influence of Modified Shuyu Pill with effects of invigorating and nourishing essence and blood on hippocampal neurons and phosphate serine-threonine kinase(p-Akt)in cerebral ischemia rats. Methods: 40 Wistar rats were divided into the normal group,sham operation group,model group and Chinese medicine group. Rat model of cerebral ischemia was established by modified permanent occlusion of bilateral common carotid artery in the model group and Chinese medicine group, and the common carotid artery was not ligated in the sham operation group. The Chinese medicine group was treated with Modified Shuyu Pill at daily dose of 10 g/kg by intragastric administration,and the other groups were treated with equivalent 0.9%NaCl solution. After administration for 6 weeks,the samples were collected. The morphological changes of hippocampus were observed by HE staining,the expression of neuronal nuclei(NeuN)was detected by immunofluorescence and the expression of p-Akt was detected by Western blot. Results: The results of HE staining showed that,the cell density was decreased in hippocampus of rats in the model group,the vertebral cell layer became thinner,the cells arranged loosely with large intercellular space,and the nuclear atrophy was observed. After the intervention of Modified Shuyu Pill,the cell density was increased in hippocampus of rats in the Chinese medicine group,the vertebral cell layer became thicker,the cells arranged closely with small intercellular space,and the nuclear atrophy was reduced. Compared with the sham operation group,the expression of p-Akt in hippocampus was obviously decreased(P<0.05)and the expression of NeuN was obviously increased(P<0.05)in the model group. Compared with the model group,the expressions of NeuN and p-Akt in hippocampus were obviously increased in the Chinese medicine group(P<0.05). Conclusion: Modified Shuyu Pill can promote the expressions of NeuN and p-Akt in hippocampus of rats with cerebral ischemia,and may promote the repair of hippocampal neuronal damage caused by cerebral ischemia and inhibit the apoptosis of neurons.
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