基于RNA-Seq高通量测序分析脑心通胶囊抑制炎症反应转录组学特征

Analysis on transcriptionomic characteristics of Naoxintong Capsules in prevention of post-ischemic inflammation based on RNA-Seq technology

该研究采用高通量测序方法探讨脑心通胶囊抑制脑缺血后炎症反应的作用机制。首先,建立了1.0μg·mL^-1 LPS诱导小胶质细胞BV-2炎症模型,考察了不同浓度的脑心通胶囊肠吸收液(62.5,31.25,15.63,7.81μg·mL^-1)对LPS诱导小胶质细胞BV-2炎症因子分泌的影响;继而采用RNA-Seq高通量测序检测脑心通胶囊干预LPS诱导BV-2细胞中差异表达基因,GO和KEGG对差异表达的基因进行富集分析,筛选脑心通胶囊抑制炎症反应的潜在生物学过程及相关信号通路。结果表明,不同浓度的脑心通胶囊肠吸收液能够显著抑制LPS诱导BV-2中炎症介质的释放,且随着浓度升高抑制作用呈明显的量效关系。高通量测序结果显示,脑心通胶囊逆转的差异表达基因392个,这些差异表达基因主要通过参与Toll样受体信号通路、趋化因子信号通路、TNF信号通路等炎症相关信号通路在脑心通胶囊抑制炎症中发挥重要作用。该研究表明,脑心通胶囊能够抑制脑缺血后的炎症反应,且作用机制与调控Toll样受体信号通路、趋化因子信号通路、TNF信号通路等相关。

In this research,high-throughput sequencing was used to investigate the mechanism of Naoxintong Capsules(NXTC)in prevention of post-ischemic inflammation.First,microglia BV-2 inflammatory model was induced by 1.0μg·mL^-1 LPS to investigate the effect of intestinal absorption solution of NXTC(NXTCIA)at different concentrations(62.5,31.25,15.63,7.81μg·mL^-1)on LPS-induced BV-2 inflammatory factors in microglia.Then,an RNA-Seq high-throughput sequencing method was performed to identify the differentially expressed mRNAs in microglia BV-2 after pre-treatment with NXTC.GO and KEGG enrichment analysis was used to screen the potential biological processes and related signaling pathways of NXTC in inhibiting inflammation.The results showed that four NXTCIA concentrations could significantly inhibit the release of LPS-induced inflammatory mediators in BV-2 in a dose-dependent manner.Furthermore,high-throughput sequencing results showed that 392 mRNA transcripts were reversed following pre-treatment with NXTC.GO enrichment analysis showed that the transcripts reversed by NXTC were mainly involved in Toll-like receptor signaling pathway,chemokine signaling pathway,and TNF signaling pathway.Taken together,our findings showed that NXTC treatment could provide protective effects against inflammatory response and the mechanism might be related to the regulation of Toll-like receptor signaling pathway,chemokine signaling pathway,and TNF signaling pathway.