基于转录组高通量测序探讨清解化攻方抑制重症急性胰腺炎大鼠炎症反应的机制

Mechanism of Qingjie Huagong Decoction inhibiting inflammatory response in rats with severe acute pancreatitis based on transcriptome high-throughput sequencing

目的:采用转录组高通量测序探讨清解化攻方抑制重症急性胰腺炎(SAP)炎症反应的可能机制。方法:雨蛙素联合脂多糖腹腔注射法复制SAP大鼠模型,观察空白组、模型组、中药组大鼠胰腺组织湿/干比重、血清淀粉酶、内毒素、IL-6、IL-8表达水平及病理组织改变情况;RNA-seq测序检测大鼠胰腺组织基因表达差异,筛选清解化攻方抑制炎症反应的核心基因、生物学功能及信号通路;Real-time PCR验证测序结果。结果:与模型组比较,中药组大鼠胰腺湿/干比重显著下降(P<0.01);ELISA及胰腺病理结果表明中药组大鼠胰腺水肿、炎性因子释放以及胰腺炎症反应显著改善。测序表明中药组上调1701个、下调1602个胰腺差异基因,从而调节免疫炎症反应、趋化因子介导通路等生物过程,并下调TNF-α/NF-κB、MAPK、趋化因子等信号通路。中药组TNF-α、NF-κB p65、IKKβ、p38MAPK、ERK1、IL-1β较模型组表达显著下降,IκBα表达显著增加,与测序结果一致。结论:清解化攻方能有效抑制重症急性胰腺炎大鼠炎症反应,可能机制与调节TNF-α/NF-κB、MAPK等信号通路相关。

Objective:To explore the possible mechanism of Qingjie Huagong Decoction(QJHGD)inhibiting the inflammatory response of severe acute pancreatitis(SAP)by transcriptome high-throughput sequencing.Methods:The SAP rat model was established by intraperitoneal injection of cerulein combined with lipopolysaccharide.The wet/dry specific gravity of pancreatic tissue,the expression levels of serum amylase,endotoxin,IL-6 and IL-8 and the pathological changes were observed in the blank group,model group and Chinese medicine group;RNA-Seq sequencing was used to detect the difference of gene expression in rat pancreas,and to screen the core genes,biological functions and signal pathways of QJHGD inhibiting inflammatory response;The results were verified by Real-time PCR.Results:Compared with the model group,the wet/dry ratio of pancreas in Chinese medicine group decreased significantly(P<0.01).The results of ELISA and pancreatic pathology showed that the pancreatic edema,the release of inflammatory factors and the response to pancreatitis were significantly improved in Chinese medicine group.Sequencing showed that the Chinese medicine group up-regulated 1701 and down-regulated 1602 pancreatic differential genes,so as to regulate biological processes such as immune inflammatory response and chemokine mediated pathway,and down-regulate TNF-α/NF-κB.MAPK,chemokine and other signal pathways.The expression of TNF-α,NF-κB p65,IKKβ,p38MAPK,ERK1,IL-1βin the Chinese medicine group was significantly lower than that in the model group,IκBαwas contrary.The above results are consistent with the sequencing results.Conclusion:QJHGD can effectively inhibit the inflammatory response in rats with severe acute pancreatitis,which may be related to the regulation of TNF-α/NF-κB,MAPK and other signal pathways.