目的 通过转录组测序技术（RNA sequencing，RNA-seq）分析天麻总酚（total polyphenols of Gastrodia elata，TPGE）抗大鼠脑缺血再灌注损伤（cerebral ischemic reperfusion injury，CIRI）的转录组学特征，并采用定量聚合酶链反应（quantitative polymerase chain reaction，qPCR）验证其分子作用机制。方法 将动物分为假手术组、模型组及TPGE组（262.3 mg/kg），通过线栓法复制大鼠脑中动脉栓塞/再灌注损伤（middle cerebral artery embolization/reperfusion injury，MCAO/R）模型，连续ig给药14 d后，以神经行为学评分评价TPGE对MCAO/R大鼠的脑保护作用；采用RNA-seq技术对大鼠脑组织进行转录组测序分析，筛选出TPGE抗CIRI的相关差异基因，对其进行基因本体（gene ontology，GO）和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）分析；通过qPCR对筛选出的差异基因进行实验验证。结果 神经行为学评分提示TPGE可明显改善MCAO/R大鼠的神经功能缺失症状；转录组学数据表明TPGE主要作用于Itgam、Tnfsf13b、Itgb2、Cd86、Cxcl16、Cxcl10、Irf7、Tlr4、Csf2rb、Csf3r等差异基因，GO分析主要富集在炎症反应、免疫应答和应激反应等生物学过程，KEGG分析主要富集在炎症、凋亡、肿瘤坏死因子（tumor necrosis factor，TNF）和Toll样受体等信号通路；qPCR结果表明，与模型组比较，TPGE组大鼠脑组织中的MyD88、NF-κB、TNF-α、MCP-1、IL-1β、P62、LC-3、LC3-I/Ⅱ、Bax mRNA表达水平显著降低，差异具有统计学意义（P<0.05、0.01、0.001）。结论 TPGE具有抗大鼠CIRI的作用，转录组学特征提示其抗CIRI的关键差异基因主要与抑制炎症、减轻自噬和抗凋亡反应相关，可能通过干预MyD88/NF-κB/TNF-α信号通路发挥作用。

Objective To analyze the transcriptome characteristics of total polyphenols of Gastrodia elata (TPGE) against cerebral ischemia-reperfusion injury (CIRI) in rats via RNA-Seq high-throughput sequencing. Quantitative polymerase chain reaction (qPCR) was used to verify the molecular mechanism. Methods The animals were divided into sham group, model group, and TPGE group (262.3 mg/kg). The model of middle cerebral artery occlusion/reperfusion injury (MCAO/R) in rats was established by thread occlusion. After continuous ig administration for 14 d, the neurobehavioral score, Hematoxylin-Eosin (HE) staining, and Nissl staining were utilized to evaluate the neuroprotective effects of TPGE on CIRI rats. The transcriptome sequencing of the brain tissue of rats in each group was carried out by RNA-Seq technique. The differential expressed genes related to anti-CIRI of TPGE were screened and analyzed by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG). Results Neurological Severity Scores (NSS) showed that TPGE can improve the pathological damage of neurons in brain tissue of MCAO/R. The analysis of transcriptome data showed that TPEG mainly effected on Itgam, Tnfsf13b, Itgb2, Cd86, Cxcl16, Cxcl10, Irf7, Tlr4, Csf2rb, and Csf3r. GO analysis was mainly concentrated in inflammatory response, immune response, stress response, and regulation of tumor necrosis factor; KEGG analysis was mainly concentrated in chemokine signaling pathway, NOD-like receptor signaling pathway, Toll-like receptor signaling pathway, and TNF signaling pathway. The qPCR results showed that, compared with model group, the mRNA exprssion level of MyD88, NF-κB, TNF-α, MCP-1, IL-1β, P62, LC-3, LC3-I/Ⅱ, and Bax in brain tissue of rats in TPGE group significantly reduced. The difference was statistically significant (P<0.05, 0.01, 0.001). Conclusion TPGE has anti-CIRI effect in rats, its mechanism may be related to the regulation of chemokine, Toll-like receptors, NOD-like receptors, and TNF signaling pathways.

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国家自然科学基金项目（81660677）；云南省科技计划中医联合重点项目[2018FF001（-007）]；云南省科技计划中医联合重点项目[2017FF117（-004）]；云南省科技计划重点项目（2017FA046）