基于网络药理学及实验验证探讨四君子汤抑制慢性结直肠炎腺瘤发生的作用机制

Exploring the Mechanism of Sijunzi Decoction in Inhibiting the Development of Adenoma in Chronic Colorectal Inflammation Based on Network Pharmacology and Experimental Validation

目的通过网络药理学预测并利用动物实验验证四君子汤的核心靶点,探究四君子汤抑制慢性结直肠炎腺瘤发生的作用机制。方法基于TCMSP、GeneCards及OMIM数据库,筛选四君子汤活性成分潜在靶标以及疾病与免疫相关基因靶点,构建四君子汤活性成分潜在靶标调控网络,通过通路富集和蛋白互作网络分析相关通路和核心靶点,探究四君子汤抑制肠道疾病的潜在作用靶点。通过对DSS诱导的慢性结直肠炎小鼠模型进行四君子汤干预,观察各组小鼠肠道表型及病理染色的变化,证实四君子汤对慢性结直肠炎小鼠的保护作用。最后,通过qRT-PCR及Western blot验证了基于网络药理学预测的与四君子汤相关的核心靶点的转录水平和蛋白的表达水平,进一步验证四君子汤抑制慢性结直肠炎小鼠腺瘤发生的作用机制。结果网络药理学分析表明,四君子汤与炎症、免疫和肿瘤相关通路有关,并筛选预测出15个相关的核心靶点。四君子汤干预DSS慢性结直肠炎模型小鼠后,减缓了结直肠的炎症反应,对肠道肿瘤的数量和大小均有显著改善。qRT-PCR和Western blot则进一步验证了四君子汤的作用机制与网络药理学预测的核心靶点EGFR、MAPK8和CASP3有关。结论四君子汤能够抑制EGFR、MAPK8和CASP3的表达水平,抑制慢性结直肠炎腺瘤发生,为进一步深入探究四君子汤抑制慢性结直肠炎腺瘤发生的作用机制提供了新思路。

Objective To predict and validate the core targets of Sijunzi Decoction through network pharmacology and animal experiments,and to explore the mechanism of action of Sijunzi Decoction in inhibiting the occurrence of adenomas in chronic colorectal inflammation.Methods Based on TCMSP,GeneCards and OMIM databases,screen potential targets for active components of Sijunzi Decoction and gene targets related to disease and immunity,construct a potential target regulatory network for active components of Sijunzi Decoction,analyze relevant pathways and core targets through pathway enrichment and protein interaction networks,and explore potential targets of Sijunzi Decoction in inhibiting intestinal diseases.By intervening in the DSS induced chronic colorectal inflammation mouse model with Sijunzi Decoction,the protective effect of Sijunzi Decoction on chronic colorectal inflammation mice was confirmed through changes in intestinal phenotype and pathological staining in each group of mice.Finally,the transcriptional levels and protein expression levels of the core targets related to Sijunzi Decoction predicted based on network pharmacology were verified through qRT PCR and Western Blot,further validating the mechanism of Sijunzi Decoction′s inhibitory effect on the occurrence of adenomas in mice with chronic colorectal inflammation.Results Network pharmacology analysis shows that Sijunzi Decoction is related to inflammatory,immune,and tumor related pathways,and 15 related core targets have been screened and predicted.After intervention with Sijunzi Decoction in DSS chronic colorectal inflammation model mice,it slowed down the inflammatory response of the colon and significantly improved the number and size of intestinal tumors.QRT PCR and Western Blot further confirmed that the mechanism of action of Sijunzi Decoction is related to the core targets of network pharmacology prediction,EGFR,MAPK8,and CASP3.Conclusion Sijunzi Decoction can inhibit the expression levels of EGFR,MAPK8,and CASP3,exerting the effect of inhibiting the occurrence of adenomas in chronic colorectal inflammation.This provides a new idea for further exploring the mechanism of Sijunzi Decoction′s inhibition of the occurrence of adenomas in chronic colitis.