外源质粒DNA对小鼠脾脏物质能量代谢的影响

研究外源质粒DNA经胃肠道途径对免疫激活状态下小鼠脾脏代谢的影响。给Balb/c小鼠灌喂质粒pcDNA3 200μg，分别在灌喂后4h和18h分离脾脏，提取脾脏总RNA。利用Affymetrix基因表达谱芯片对灌喂质粒pcDNA3后的Balb/c小鼠脾脏进行基因表达谱研究。采用Genmmapp和MAPPFinder软件进行功能聚类分析。对上调倍数两倍及两倍以上的基因进行功能聚类发现，灌喂后4h,外源质粒刺激小鼠脾脏产生免疫应答的同时，脾脏中嘌呤代谢及蛋白质合成，胆固醇合成、脂肪酸合成、糖酵解、三羧酸循环及线粒体氧化磷酸化途径等大量代谢过程受到诱导。在灌喂后18h也得到类似的结果。结果表明外源质粒DNA可通过胃肠道途径吸收影响小鼠脾脏物质能量代谢过程。结果有助于在分子水平上深入了解外源质粒DNA经胃肠道吸收后的作用机制。

Effect of foreign plasmid DNA on material and energy metabolism of spleen in mice

To study the influence of foreign plasmid DNA on spleen metabolism in immune stimulated mice via the gastrointestinal tract. Mice were oral administered by pipette with 200μg of plasmid pcDNA3 and spleen were isolated at 4h and 18h after oral administration. Total RNA were extracted from spleen and spleen gene expression profile of Balb/c mice was analyzed by using Affymetrix oligonucleotide genechip after oral plasmid pcDNA3 administration. Functional cluster analysis was conducted by Genmapp and MAPPFinder software. By functional cluster analyzing the genes which were up regulated more than twofolds, Genmapp results showed that plenty of metabolic pathway were induced in spleen after oral administration of plasmid pcDNA3. These metabolic process included purine metabolism, pyrimidine metabolism, protein synthesis, cholesterol synthesis, fatty acid synthesis, Glycolysis, TCA cycle and mitochondria oxidative phosphorylation pathway. The similar results also took place at 18h after oral administration. The result indicated that foreign plasmid DNA can modulate metabolism process in spleen of mice via the gastrointestinal tract, and may help understand the mechanism of action of foreign plasmid DNA uptaked via the gastrointestinal tract.