加味脑泰方对血管性痴呆模型大鼠海马组织mRNA表达谱的微阵列分析

Differential mRNA expression profiles of supplemented Naotaifang in vascular dementia rats by microarray analyses

目的采用微阵列技术分析加味脑泰方干预前后血管性痴呆(VD)模型大鼠海马组织mRNA表达谱,以探究其治疗VD的分子机制。方法采用双侧颈总动脉结扎术制作VD大鼠模型,给予加味脑泰方治疗30 d,HE染色和水迷宫实验评价治疗效果,Agilent mRNA表达谱芯片获取加味脑泰方干预前后VD模型大鼠海马组织mRNA表达数据,微阵列分析筛选显著差异表达的基因,构建蛋白与蛋白相互作用(PPI)网络,GO和Pathway富集分析差异基因主要参与的生物学过程和信号通路,利用免疫组化和实时荧光定量PCR(qRT-PCR)验证芯片分析结果。结果HE染色和水迷宫实验显示VD大鼠呈现脑缺血、海马神经细胞损伤、空间学习记忆能力下降,加味脑泰方可以部分逆转该现象。微阵列技术分析筛选出加味脑泰方干预前后差异表达的基因469个,其中上调180个,下调289个,IL6、FGF2、TNF、IL1b等可能是其治疗VD的主要药效靶点,免疫组化和qRT-PCR验证了该分析结果。GO和Pathway富集分析显示,这些基因与炎症反应、凋亡过程等生物学过程密切相关,主要参与了TNF信号通路、Toll样受体信号通路、细胞凋亡途径等的调控。结论加味脑泰方对VD模型大鼠的治疗作用是多基因多途径共同参与调控的过程,抑制海马神经炎症可能是其抗VD的重要机制之一。

Objective To explore the molecular mechanism of the supplemented Naotaifang(sNTF)in the treatment of vascular dementia(VD),and the mRNA expression profiles of hippocampal tissue of VD model rats before and after the intervention of modified sNTF were investigated by microarray analysis.Methods VD model was established by bilateral common carotid artery ligation.VD rats were treated with sNTF for 30 days.HE staining and Morris water maze were used to evaluate the therapeutic effect of sNTF.The mRNA expression profiles data of VD model rats before and after intervention of sNTF were obtained by Agilent mRNA expression chip.The significantly differentially expressed genes were screened by microarray analysis,and the protein-protein interaction(PPI)network was constructed.The biological processes and signaling pathways in which differentially expressed genes were mainly involved and analyzed by GO and pathway enrichment.Immunohistochemistry and qRT-PCR were used to verify the chip analysis results.Results HE staining and Morris water maze experiments showed that VD rats showed cerebral ischemia,hippocampal neuron damage,and decreased spatial learning and memory function,but sNTF can partially reverse this trend.469 differential expression genes were screened by microarray analysis,including 180 up-regulated genes and 289 down-regulated genes.IL6,FGF2,TNF,and IL1b may be the main pharmacodynamic targets of sNTF in the treatment of VD rats,and the results were verified by immunohistochemistry and qRT-PCR.GO and pathway enrichment analysis showed that these genes were closely related to biological processes such as inflammation and apoptosis,and these genes were mainly involved in the regulation of TNF signaling pathway,toll-like receptor signaling pathway and the apoptosis pathway.Conclusion The results suggested that the therapeutic effect of DSS on AD involves multiple genes and pathways,and and inhibition of hippocampal neuroinflammation may be one of the important mechanisms of its anti-VD.