杞菊地黄汤对MNU诱发的大鼠视网膜变性基因表达谱的影响

Retinal Gene Expression Profiles of MNU-induced Rat Retinal Degeneration Treated with Qijudihuangtang

目的:观察杞菊地黄汤防治MNU诱发大鼠视网膜变性的基因表达谱变化,探索其作用的分子机制。方法:30只大鼠分为3组:药物组(杞菊地黄汤灌胃+MNU皮下注射)、模型组(生理盐水灌胃+MNU皮下注射)和正常组(生理盐水灌胃)。造模后12h,取视网膜进行基因芯片分析和RT-PCR检测。并用GO(gene ontology)分类和信号通路分析等方法对差异表达的基因进行分析。结果:模型组/正常组和模型组/药物组中分别有75个和118个差异表达基因,这些基因多与信号转导、发育、免疫和防御、凋亡等生物过程有关,主要涉及到MAPK、Toll样受体和凋亡信号通路。结论:在MNU诱导的大鼠视网膜变性中,杞菊地黄汤能特异性拮抗数个信号通路中的基因表达。

Purpose： Our previous studies demonstrated that Qijudihuangtang, a compound of traditional Chinese medicine, protects from MNU （N-methy-N-nitrosourea）-induced rat retina photoreceptor injury. The current study was aimed to investigate protective effect of Qijudihuangtang on the gene expression of retina in the same model. Methods： Thirty 46-day-old female Sprague-Dawley rats were randomly divided into three groups （each group, n=10）. The rats received a daily intragastric administration of Qijudihuangtang in drug group, and 0.9% NaCl in model and normal groups for 4 days. On the fifth day, the rats in model and drug groups received a single intraperitonal injection of 40 mg/kg body weight of N-methy-N-nitrosourea Equivalent volume of 0.9% NaCl was injected to the rats of normal group. （MNU）. After 12 h of injection, all animals were sacrificed. Fresh retinas were used to extract total RNA, which was assayed by microarray and real time RT-PCR. The differentially expressed genes were also analyzed for the functional annotation and signal network mapping. Results： There were 75 and 118 genes differently expressed （ratio ≥2.0） in model group versus normal group and model group versus drug group, respectively. The former mainly included genes upregulated downregulated and the latter included most genes . These genes were mainly assigned to the categories binding transcription factor, signaling molecular, receptor, nucleic binding and extracellular matrix, and involved biological processes including signal transduction, development, immune and defense, apoptosis. The differential expressed genes contributed to MAPK signaling pathways, Toll-like receptor signaling pathway and apoptosis pathway. Conclusion： The gene expression in the early stage of MNU-induced rat retinal degeneration can be regulated significantly by the administration of Qijudihuangtang. These changed genes were associated with specific functional groups.