芫根调节免疫功能的小肠转录组学研究

Small intestinal transcriptome study on immune function regulation of Coriaria Myra

目的利用生物信息学技术初步探索芫根调控肠道免疫功能的分子生物学机制。方法选择雄性SPF级BALB/c小鼠18只,随机分为3组,每组6只。SC1组小鼠每只每次灌胃芫根提取液150μL,SC2组小鼠每只每次灌胃绞碎芫根原浆悬液150μL,NC组小鼠每只每次灌胃生理盐水150μL,连续7 d每日灌胃一次。分别取每组小鼠小肠组织样品提取RNA,总RNA的质检合格后,进行转录组测序。按GO功能和KEGG信号通路对差异表达基因聚类分析揭示差异表达高度富集的免疫相关通路。从免疫角度分析芫根灌胃小鼠小肠组织的基因表达变化情况。结果转录组测序共检测到27733条mRNA的表达,SC1组与NC组比较,显著差异表达基因1635个,其中上调差异表达基因1236个,下调差异表达基因399个;SC2组与NC组比较,显著差异表达基因2872个,其中上调差异表达基因2233个,下调差异表达基因639个(P<0.05)。按GO功能和KEGG信号通路聚类分析显示差异表达基因在白介素分泌、干扰素反应、T细胞受体信号通路及维生素和脂肪消化吸收等途径的富集度在两个芫根组中均居于前20位(P<0.01),肠黏膜趋化因子CCL20和巨噬细胞极化标志物CD274等免疫蛋白编码基因差异表达显著且同属于上述多个免疫通路。结论芫根灌胃小鼠小肠的差异表达基因在白介素分泌、干扰素反应、T细胞受体信号通路、营养物质消化吸收等途径高度富集,提示芫根对肠道免疫系统及肠黏膜功能的调节,其中CD274的表达上调和CCL20的表达下调提示诱导M1型巨噬细胞极化和T细胞活化可能是芫根调控免疫和维护肠道正常结构功能的重要途径。

Objective Using bioinformatics technology,the molecular biological mechanism of Brassica rapa regulating intestinal immune function was explored.Methods Male BALB/C mice were randomly divided into three groups,with 6 mice in each group.SC1 group was gavaged with Brassica rapa extract,SC2 group was gavaged with chopped Brassica rapa slurry suspension,and NC group was gavaged with normal saline once a day for 7 consecutive days as blank control.RNA was extracted from small intestine tissue samples of each group,and transcriptome sequencing was performed after the total RNA was qualified.Cluster analysis of differentially expressed genes was performed according to GO function and KEGG signaling pathway,and mapping revealed immune-related pathways with highly enriched differentially expressed genes.The changes of gene expression in small intestine of Brassica rapa sinensis mice were analyzed from immune Angle.Results The expression of 27733 mRNA was detected by transcriptome sequencing.Compared with NC group,there were 1635 differentially expressed genes in SC1 group,including 1236 differentially expressed genes up-regulated and 399 differentially expressed genes down-regulated.Compared with NC group,2872 genes were significantly differentially expressed in SC2 group,including 2233 up-regulated genes and 639 down-regulated genes.Cluster analysis of GO function and KEGG signaling pathway showed that the enrichment degree of differentially expressed genes in interleukin secretion,interferon response,T cell receptor signaling pathway and vitamin and fat digestion and absorption were among the top 20 in both Brassica rapa groups.Immunoprotein-coding genes such as intestinal mucosa chemokine CCL20 and macrophage polarization marker CD274 were significantly differentially expressed and belonged to the above immune pathways(P<0.05).Conclusion The differentially expressed genes in the small intestine were highly enriched in interleukin secretion,interferon response,T cell receptor signaling pathway and nutrient digestion and absorption,suggesting that The function of intestinal immune system and intestinal mucosa was regulated by Brassica rapa.The up-regulated expression of Cd274 and down-regulated expression of Ccl20 suggest that the induction of polarization of M1-type macrophages and activation of T cells may be an important way to regulate the immunity and maintain the normal intestinal structure and function.