基于巨噬细胞M2极化探讨胃黏膜肠上皮化生的发生机制及中药预测研究

Mechanism of Intestinal Metaplasia of Gastric Mucosa and Prediction of Traditional Chinese Medicine Based on M2 Polarization of Macrophages

目的探讨巨噬细胞M2极化对胃黏膜肠上皮化生(IM)的影响及治疗靶点,为中医药防治IM提供新思路和新方向。方法从公共数据库下载IM及巨噬细胞M2极化相关数据集;采用免疫浸润分析、基因相关性分析、功能富集分析、蛋白互作网络及免疫荧光等方法进一步阐述IM及巨噬细胞M2极化的相关性;最后将获得的枢纽基因与Coremine Medical平台相互映射,预测中医治疗IM的有效中药和治法。结果免疫浸润及免疫荧光分析显示巨噬细胞M2极化标记物CD68^(+)CD163^(+)(P=0.0394)、CD68^(+)CD206^(+)(P=0.002)在IM组中的表达均显著高于正常组,且M2型巨噬细胞在IM组的的浸润水平显著高于正常组(P<0.05)。IM相关标记基因(CDX1、MUC2、TFF3)与巨噬细胞M2极化标志物(CD209、ARG1、MSR1、STAT3、IL32、SELENOP)的表达水平显著正相关(P<0.05);功能富集分析提示IM与巨噬细胞M2极化共表达的差异基因主要与羧酸跨膜转运蛋白活性、有机酸跨膜转运蛋白活性等分子生物学功能密切相关;最后筛选出IM与巨噬细胞M2极化共表达的枢纽差异基因7个,分别为TRAF1、TNFRSF12A、BIRC3、TNFRSF11A、CTSV、SLC29A1和CDA;中药预测共映射出中医治疗IM的中药36味,按照功效分类可归为21类,其中清热解毒药出现频次最高,其次为补气药和活血止痛药等。结论巨噬细胞M2极化可能通过外泌体途径参与IM病理的发生及发展过程,IM与巨噬细胞M2极化共表达的7个枢纽基因(TRAF1、TNFRSF12A、BIRC3、TNFRSF11A、CTSV、SLC29A1和CDA)和IM特异性分子CDX1、MUC2、TFF3可能存在相互关联作用,共同参与IM进展;“健脾化瘀解毒法”是中医治疗IM的核心治法,从巨噬细胞M2极化出发有望成为临床防治IM的新方向和突破口。

Objective To explore the effect of M2 polarization of macrophages on intestinal metaplasia(IM)of gastric mucosa and its therapeutic target,so as to provide new ideas and directions for the prevention and treatment of IM with traditional Chinese medicine.Methods The data sets related to IM and macrophage M2 polarization were downloaded from the public database,and the correlation between IM and macrophage M2 polarization was further expounded by immune infiltration analysis,gene correlation analysis,functional enrichment analysis,protein interaction network and experimental verification.Finally,the obtained hub genes were mapped with Coremine Medical platform to predict the effective traditional Chinese medicine and treatment of IM.Results Immunoinfiltration and immunofluorescence analysis showed that the expression of macrophage M2 polarization markers CD68^(+)CD163^(+)(P=0.0394)and CD68^(+)CD206^(+)(P=0.002)in IM group was significantly higher than that in normal group,and the infiltration level of M2 macrophages in IM group was significantly higher than that in normal group(P<0.05).IM related marker genes(CDX1,MUC2,TFF3)were positively correlated with macrophage M2 polarization markers(CD209,ARG1,MSR1,STAT3,IL32,SELENOP)(P<0.05).Functional enrichment analysis showed that the differential genes co-expressed by IM and macrophage M2 polarization were closely related to molecular biological functions such as carboxylic acid transmembrane transporter activity and organic acid transmembrane transporter activity.Finally,7 pivotal differential genes co-expressed by IM and macrophage M2 were screened,which were TRAF1,TNFRSF12A,BIRC3,TNFRSF11A,CTSV,SLC29A1 and CDA,respectively.According to the prediction of traditional Chinese medicine,23 kinds of traditional Chinese medicine for IM were predicted,which could be classified into 14 categories according to their efficacy,among which heat-clearing antidote appeared most frequently,followed by drugs for tonifying qi and painkillers for promoting blood circulation drugs.Conclusion Macrophage M2 polarization may be involved in the pathogenesis and development of IM through exocrine pathway.IM and seven hub genes co-expressed by macrophage M2 polarization(TRAF1,TNFRSF12A,BIRC3,TNFRSF11A,CTSV,SLC29A1 and CDA)and IM specific molecules CDX1,MUC2 and TFF3 may be associated with each other and participate in the progression of IM.The therapeutic method of Jianpi Huayu Jiedu is the core treatment of IM in traditional Chinese medicine.Starting from the M2 polarization of macrophages,it is expected to become a new direction and breakthrough in clinical prevention and treatment of IM.