化痰祛瘀方在肝豆状核变性模型小鼠海马神经细胞增殖中的作用及相关机制研究

目的:探讨化痰祛瘀方在肝豆状核变性(Wilson病)模型小鼠海马神经细胞增殖中的作用及PI3K-AKT-mTOR信号通路相关机制。方法:准备健康4月龄Wilson病模型TX小鼠和DL小鼠。制备中药化痰祛瘀方溶液备用。分组设立正常对照组、模型组和实验组,按实验设计分组处理。采用Western blot检测PI3K-AKT-mTOR信号通路相关因子和增殖相关因子的蛋白表达水平; 同时,采用Brd U掺入法检测化痰祛瘀方处理Wilson病模型小鼠海马神经细胞增殖。结果:与正常对照组相比,模型组总PI3K、AKT和mTOR及p-PI3K、p-AKT和p-mTOR蛋白水平均明显上调,而PCNA和Ki-67蛋白表达水平及Brd U阳性细胞数明显降低(均P<0.05)。同时,与模型组相比,实验组总PI3K、AKT和mTOR蛋白水平及p-PI3K、p-AKT和p-mTOR均明显降低,而PCNA和Ki-67蛋白表达水平和Brd U阳性细胞数明显增加(均P<0.05)。结论:化痰祛瘀方可有助于促进Wilson病模型小鼠海马神经细胞增殖,增加PCNA和Ki-67蛋白表达水平,其可能与抑制PI3K-AKT-mTOR信号通路激活,抑制PI3K、AKT和mTOR磷酸化水平相关。

Effect and mechanism of Huatan Quyu recipe on proliferation of hippocampal neurons in mice with Wilson disease

Objective:To investigate the effect of Huatan Quyu recipe on the proliferation and invasion ability of breast cancer MCF-7 cell line and its impact on the expression level of microRNA-100(miR-100).Methods:Healthy 4-month-old TX mice and DL mice were purchased for experiments.Huatan Quyu recipe solution was prepared for subsequent usage.The normal control group,model group and experimental group were set up.Western blot was used to detect the protein expression levels of factors related to PI3K-AKT-mTOR signaling pathway and cell proliferation; meanwhile,Brd U incorporation method was used to detect the proliferation of hippocampal nerve cells in Wilson disease model mice treated with Huatan Quyu recipe.Results:Compared with the normal control group,the protein expressions of total PI3K,Akt and mTOR,p-PI3K,p-Akt and p-mTOR were significantly upregulated,while those of PCNA and Ki-67 as well as the number of Brd U positive cells were reduced obviously in the model group(all P<0.05).At the same time,compared with the model group,the protein expression levels of total PI3K,Akt and mTOR,p-PI3K,p-AKT and p-mTOR were evidently decreased,while those of PCNA and Ki-67 as well as the number of Brd U positive cells were highly increased(all P<0.05).Conclusion:Huatan Quyu recipe can promote the proliferation of hippocampal neurons and increase the expression levels of PCNA and Ki-67 protein in Wilson disease model mice,which may be related to the inhibited activation of PI3K-Akt-mTOR signaling pathway and the suppressed phosphorylation levels of PI3K,Akt and mTOR.