天麻总酚抗大鼠脑缺血再灌注损伤的转录组学分析及实验验证

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(...

Analysis on transcriptomic characteristics of total phenols of Gastrodia elata in prevention of CIRI based on RNA-seq technology

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,Tnfsf13 b,Itgb2,Cd86,Cxcl16,Cxcl10,Irf7,Tlr4,Csf2 rb,and Csf3 r.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 My D88,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.