目的 通过观察氯胺酮麻醉对大鼠大脑皮质转录组表达的影响，探讨其可能的麻醉机制。 方法 选择5~6周龄雄性SD大鼠6只，采用完全随机法分为氯胺酮组（KET组）和对照组（Ctrl组），每组3只。分别腹腔注射麻醉剂量氯胺酮（50 mg/kg）或等体积生理盐水后30 min，提取大脑皮质进行mRNA测序（mRNA sequencing, RNA‑seq），筛选差异表达基因。使用实时荧光定量聚合酶链反应（real time fluorescent quantitative polymerase chain reaction, RT‑qPCR）进行验证，并通过基因本体（gene ontology, GO）分析、京都基因和基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路分析和蛋白质‑蛋白质相互作用（protein‑protein interaction, PPI）网络分析鉴定差异表达基因涉及的相关功能变化。 结果 在确定样本质量合格后，高通量测序检测出氯胺酮麻醉后大鼠皮质中70种基因显著上调，83种基因显著下调。通过GO、KEGG和PPI分析发现，氯胺酮麻醉过程中发生显著变化的通路主要有5‑羟色胺能突触、核糖体和光传导3种，分别对应5‑羟色胺系统、神经内分泌系统和视觉系统。 结论 氯胺酮麻醉后大鼠大脑皮质中mRNA表达显著改变。相关的功能变化分析提示，5‑羟色胺系统、神经内分泌系统和视觉系统可能参与了氯胺酮的麻醉调控。 结论：氯胺酮麻醉后大鼠大脑皮层中mRNA表达显著改变。相关的功能变化分析提示，5-羟色胺系统、神经内分泌系统和视觉系统可能参与了氯胺酮的麻醉调控。
Objective To observe the effect of ketamine anesthesia on the expression of transcriptome in rat brain tissue and to explore its possible anesthetic mechanism. Methods A total of 6 SD rats (male, aged 5‒6 weeks) were divided into two groups according to complete randomization (n=3): a ketamine group (KET group) and a control group (Ctrl group). Then, 30 min after intraperitoneal injection of ketamine (50 mg/kg) or the same volume of normal saline, the cerebral cortex was collected for mRNA sequencing (RNA‑seq) to screen differentially expressed genes. Real‑time fluorescent quantitative polymerase chain reaction (RT‑qPCR) was used for verification. The functional changes of differentially expressed genes were identified through gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and protein‑protein interaction (PPI) analysis.. Results The quality of the samples were confirmed. High‑throughput sequencing results showed that 70 genes were significantly up‑regulated and 83 genes were significantly down‑regulated in the cortex of rats after ketamine anesthesia. GO, KEGG and PPI analyses indicated that the pathways that significantly changed during ketamine anesthesia included 5‑serotonergic synapse, ribosome and photoconduction corresponding to 5‑hydroxytryptamine system, neuroendocrine system and visual system respectively. Conclusions The expression of mRNA in rat brain significantly change after ketamine anesthesia. The analysis of related functional changes suggests that 5‑serotonin system, neuroendocrine system and visual system may be involved in the anesthetic regulation of ketamine. Results After confirming that the quality of the sample, 70 significantly up-regulated genes and 83 significantly down-regulated genes in the brain tissue of rats after ketamine anesthesia were identified. GO, KEGG and PPI analysis found that the pathways changed significantly during ketamine anesthesia mainly included serotonergic synaptic, ribosomal and photoconduction corresponding to 5-hydroxytryptamine system, neuroendocrine system and visual system respectively. Conclusion The expression of mRNA in rat brain changed significantly after ketamine anesthesia. The analysis of related functional changes suggests that the serotonin system, neuroendocrine system and visual system may be involved in the anesthetic regulation of ketamine.
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