熊胆粉改善大鼠实验性结肠炎及其作用靶点预测

Study on the effect of Bear Bile Powder on experimental colitis in rats and its target prediction

目的:探究熊胆粉对3-硝基苯磺酸(3-nitrobenzenesulfonic acid,TNBS)诱导大鼠实验性结肠炎的治疗作用,结合网络药理学和分子对接的方法,预测熊胆粉治疗炎症性肠病的药效物质、作用靶点和潜在通路。方法:将大鼠随机分正常组,模型组,阳性药组,熊胆粉高,中,低组。利用TNBS灌肠诱导大鼠实验性结肠炎,评估疾病活动指数(disease activity index,DAI)分数,测量结肠长度和病理改变,ELISA法测定结肠组织TNF-α、IL-1β、IL-6、IL-10炎症因子含量,然后通过UPLC-Q Exactive Orbitrap-MS/MS分析熊胆粉成分,利用相关数据库,预测其可能的信号通路,并对二者的亲和力进行预测。结果:与模型组相比,熊胆粉各剂量组DAI分数显著降低(P<0.01),结肠长度不同程度增加,低剂量组更为显著(P<0.05);结肠组织的炎性细胞浸润减少,黏膜上皮基本完整,腺体排列较规则。检测相关炎症因子,与模型组相比,熊胆粉低剂量组TNF-α含量显著下降(P<0.05),IL-10含量显著上升(P<0.01),熊胆粉各剂量组IL-1β、IL-6含量显著下降(P<0.05)。网络药理学和分子对接结果表明胆红素可能是熊胆粉治疗IBD的重要成分;PIK3CA、AKT1、MAPK1、MAPK14、MAPK3是治疗疾病的重要靶点;PI3K-Akt信号通路、FoxO信号通路、MAPK信号通路、EGFR酪氨酸激酶抑制剂抗性、HIF-1信号通路则是治疗疾病的重要信号通路。结论:熊胆粉能减轻TNBS诱导SD大鼠实验性结肠炎,其中胆红素可能为熊胆粉治疗IBD的关键成分,其作用机制可能与抑制炎症、诱导肠黏膜自噬、肠道免疫相关。

OBJECTIVE To explore the therapeutic effect of Bear Bile Powder on TNBS-induced experimental colitis in rats,and to predict the therapeutic substances,targets and potential pathways of bear bile powder in the treatment of inflammatory bowel disease by combining network pharmacology and molecular docking methods.METHODS The rats were randomly divided into normal group,model group,positive drug group,high,medium and low Bear Bile Powder groups.Experimental colitis was induced by TNBS enema in SD rats.Disease activity index(DAI)score,colon length and pathological changes were measured.Then the components of bear bile powder were obtained by UPLC-Q Exactive Orbitrap-MS/MS analysis,and the network pharmacology related database was used to predict the possible signaling pathways and the affinity between them.RESULTS Compared with the model group,the DAI scores of the Bear Bile Powder groups were significantly decreased(P<0.01),and the colon length was increased to varying degrees,especially in the low-dose group(P<0.05).The inflammatory cell infiltration in the colon tissue of the Bear Bile Powder groups was reduced,the colonic mucosal epithelium was basically intact,and the glands were arranged regularly.Compared with the model group,the content of TNF-α in the low-dose Bear Bile Powder group decreased significantly(P<0.05),the content of IL-10 increased significantly(P<0.001),and the content of IL-1β and IL-6 in each dose of bear bile powder groups also decreased significantly(P<0.05).The results of network pharmacology and molecular docking indicated that bilirubin may be an important component in the treatment of IBD.PIK3CA,AKT1,MAPK1,MAPK14 and MAPK3 were important targets for the treatment of diseases.PI3K-Akt signaling pathway,FoxO signaling pathway,MAPK signaling pathway,EGFR tyrosine kinase inhibitor resistance,and HIF-1 signaling pathway were important signaling pathways for the treatment of diseases.CONCLUSION Bear Bile Powder can alleviate TNBS-induced experimental colitis in SD rats.Bilirubin may be the key component of Bear Bile Powder in the treatment of IBD,and its mechanism may be related to inhibiting inflammation,inducing intestinal mucosal autophagy and intestinal immunity.