Objective To explore the active components,therapeutic targets,and molecular mechanisms of Shaoyao Decoction in the treatment of ulcerative colitis(UC)using network pharmacology techniques,bioinformatics methods,and experimental approaches.Methods Screening active components of Shaoyao Decoction and predicting their targets using databases such as PubChem,screening HCC-related disease targets through the NCBI database,constructing a PPI network,conducting GO functional enrichment and KEGG pathway analysis to identify potential biological processes and signaling pathways involved,and validating with molecular docking using AutoDock Tools.Forty male C57BL/6J mice were randomly divided into normal,model,Mesalazine,and Shaoyao Tang groups based on body weight.Except for the normal group,all other groups were induced with ulcerative colitis(UC)by providing 2.5%dextran sulfate sodium(DSS)in drinking water for 5 days.After continuous intragastric administration for 7 days,the mice were sacrificed.The levels of cytokines such as IL-2,IL-1β,IL-6,and TNF-α in colon tissues were measured by ELISA,and pathological sections of colon tissue samples were observed.Results The study identified 20 active components and 945 targets of Shaoyao Tang,among which 609 were related to UC.Through PPI network analysis,22 key targets including VEGFA,AKT1,PTGS2,and STAT3 were determined.GO analysis revealed 409 enriched terms,involving negative regulation of inflammatory response to antigenic stimulus,positive regulation of inflammatory response,etc.KEGG analysis discovered 136 significantly enriched pathways,including the NF-κB signaling pathway,Toll-like receptor signaling pathway(related to inflammation and immunity),VEGF signaling pathway,and ErbB signaling pathway(related to cell proliferation and apoptosis).Molecular docking revealed that the active ingredients exhibited strong affinity with target proteins such as IL-6,TNF,TLR4,IL-2,IL-1B,and PTGS2,forming stable conformations.The final ELISA results demonstrated that the levels of multiple inflammatory cytokines in the colon tissue of DSS-induced ulcerative colitis(UC)mouse models were significantly elevated,with notable upregulation of IL-2,IL-1β,IL-6,and TNF-α compared to the normal group(P<0.05).Following drug intervention,both the Mesalazine group and the Shaoyao Decoction group exhibited significant anti-inflammatory effects,effectively reducing the expression levels of the aforementioned inflammatory cytokines in the colon tissue(P<0.05).Notably,compared to the Mesalazine group,Shaoyao Decoction demonstrated a more pronounced regulatory effect on inflammatory cytokines(P<0.05).Conclusion In this study,the innovative integration of network pharmacology prediction,molecular docking validation and key inflammatory factor assay systematically elucidated the"multi-component-multi-target-multi-pathway"anti-inflammatory mode of Paeonia lactiflora broth in the treatment of UC.The experiments demonstrated that Paeonia lactiflora broth could regulate the release of pro-inflammatory cytokines by regulating the levels of IL-2,IL-1β,IL-6,TNF-α cytokines and other related cytokines to reduce the inflammation of the colon and improve the damage of colon tissues in mice with UC.

Xiao Chaihutang， originating from the Treatise on Typhoid and Miscellaneous Diseases， is a classic formula for harmonizing the Shaoyang. It excels in regulating the pivotal mechanism and unblocking the triple energizer， corresponding to the pathogenesis of digestive system tumors characterized by the interlocking of deficiency， stasis， phlegm， and toxicity， as well as disharmony between Yin and Yang. This paper systematically reviews research findings from China and abroad over the past decade， exploring the anti-tumor effects of Xiao Chaihutang on digestive system tumors from three dimensions： theoretical rationale， clinical efficacy， and molecular mechanisms. At the level of principle and method， Xiao Chaihutang takes "harmonization" as its core therapeutic guideline. By reconciling the exterior and interior to restore the Shaoyang pivot， harmonizing Yin and Yang to improve the tumor microenvironment， and regulating the liver and spleen to consolidate and protect the foundation of postnatal essence， it promotes the restoration of the body's dynamic balance of Yin and Yang. Clinical studies have demonstrated that Xiao Chaihutang， used alone or in combination with modern medical therapies， shows definite efficacy against digestive system tumors such as hepatocellular carcinoma， pancreatic carcinoma， and gastrointestinal carcinoma. It can significantly improve patients' quality of life， inhibit tumor progression， effectively relieve concomitant symptoms such a s cancer-related fever， anxiety， depression， and insomnia， and alleviate postoperative embolic syndromes as well as adverse reactions to radiotherapy and chemotherapy. Experimental studies have revealed that Xiao Chaihutang can inhibit tumor cell proliferation， induce apoptosis， arrest the cell cycle， suppress tumor cell invasion and metastasis， and improve the tumor microenvironment. Through the above analysis， this study elucidates the current clinical and experimental research status of Xiao Chaihutang in the treatment of digestive system tumors， aiming to provide theoretical support for its precise clinical application. On this basis， it further explores key issues in the identification of pharmacodynamic substances and the accumulation of evidence in evidence-based medicine， thereby offering a new perspective for the innovative development of integrative Chinese and Western medicine in synergistic cancer therapy.

Objective To explore the active components,therapeutic targets,and molecular mechanisms of Shaoyao Decoction in the treatment of ulcerative colitis(UC)using network pharmacology techniques,bioinformatics methods,and experimental approaches.Methods Screening active components of Shaoyao Decoction and predicting their targets using databases such as PubChem,screening HCC-related disease targets through the NCBI database,constructing a PPI network,conducting GO functional enrichment and KEGG pathway analysis to identify potential biological processes and signaling pathways involved,and validating with molecular docking using AutoDock Tools.Forty male C57BL/6J mice were randomly divided into normal,model,Mesalazine,and Shaoyao Tang groups based on body weight.Except for the normal group,all other groups were induced with ulcerative colitis(UC)by providing 2.5%dextran sulfate sodium(DSS)in drinking water for 5 days.After continuous intragastric administration for 7 days,the mice were sacrificed.The levels of cytokines such as IL-2,IL-1β,IL-6,and TNF-α in colon tissues were measured by ELISA,and pathological sections of colon tissue samples were observed.Results The study identified 20 active components and 945 targets of Shaoyao Tang,among which 609 were related to UC.Through PPI network analysis,22 key targets including VEGFA,AKT1,PTGS2,and STAT3 were determined.GO analysis revealed 409 enriched terms,involving negative regulation of inflammatory response to antigenic stimulus,positive regulation of inflammatory response,etc.KEGG analysis discovered 136 significantly enriched pathways,including the NF-κB signaling pathway,Toll-like receptor signaling pathway(related to inflammation and immunity),VEGF signaling pathway,and ErbB signaling pathway(related to cell proliferation and apoptosis).Molecular docking revealed that the active ingredients exhibited strong affinity with target proteins such as IL-6,TNF,TLR4,IL-2,IL-1B,and PTGS2,forming stable conformations.The final ELISA results demonstrated that the levels of multiple inflammatory cytokines in the colon tissue of DSS-induced ulcerative colitis(UC)mouse models were significantly elevated,with notable upregulation of IL-2,IL-1β,IL-6,and TNF-α compared to the normal group(P<0.05).Following drug intervention,both the Mesalazine group and the Shaoyao Decoction group exhibited significant anti-inflammatory effects,effectively reducing the expression levels of the aforementioned inflammatory cytokines in the colon tissue(P<0.05).Notably,compared to the Mesalazine group,Shaoyao Decoction demonstrated a more pronounced regulatory effect on inflammatory cytokines(P<0.05).Conclusion In this study,the innovative integration of network pharmacology prediction,molecular docking validation and key inflammatory factor assay systematically elucidated the"multi-component-multi-target-multi-pathway"anti-inflammatory mode of Paeonia lactiflora broth in the treatment of UC.The experiments demonstrated that Paeonia lactiflora broth could regulate the release of pro-inflammatory cytokines by regulating the levels of IL-2,IL-1β,IL-6,TNF-α cytokines and other related cytokines to reduce the inflammation of the colon and improve the damage of colon tissues in mice with UC.

[摘 要] 目的：探究miR-323a-3p、四次穿膜蛋白超家族成员1（TM4SF1）在NSCLC组织和细胞中的表达及两者间的靶向调控关系，观察两者表达对A549细胞增殖、迁移、侵袭和裸鼠移植瘤生长的影响。方法：收集2014年1月至12月间青海省人民医院手术切除的20例NSCLC组织及其相应的癌旁组织，qPCR和WB法检测癌组织中miR-323a-3p、TM4SF1 mRNA 和TM4SF1蛋白的表达。向A549细胞转染miR-323a-3p mimic，采用MTT法、Transwell法、WB法检测miR-323a-3p过表达对细胞的增殖、迁移和侵袭以及TM4SF1、细胞周期蛋白D1（cyclin D1）、p21、MMP-2、MMP-9蛋白表达的影响。采用生物信息学预测工具StarBase和双荧光素酶报告基因实验分析miR-323a-3p与TM4SF1靶向关系。将si-TM4SF1转染至A549细胞，以及分别将miR-323a-3p mimic与pcDNA或pcDNA-TM4SF1共转染A549细胞，评估细胞增殖、迁移和侵袭能力的变化；同时建立各组细胞的BALB/c裸鼠移植瘤模型，在14、21和28 d时测量并计算移植瘤体积。结果：与癌旁组织相比，NSCLC组织中miR-323a-3p表达水平明显下调，TM4SF1 mRNA和蛋白表达水平显著上调（均P<0.01）。miR-323a-3p过表达或抑制TM4SF1表达都会降低A549 细胞的增殖、迁移、侵袭能力及cyclin D1、MMP-2、MMP-9蛋白表达而促进p21蛋白表达，并且抑制A549细胞裸鼠移植瘤的生长（均P<0.01）。生物信息学StarBase工具预测和双荧光素酶基因报告实验结果显示miR-323a-3p能够靶向结合TM4SF1基因并调控 TM4SF1的表达。上调TM4SF1表达后，miR-323a-3p过表达对A549细胞恶性生物学行为及cyclin D1、MMP-2、MMP-9蛋白表达、移植瘤生长的抑制作用均被部分逆转（均P<0.01），对p21蛋白表达的促进作用也被逆转（P<0.01）。结论：miR-323a-3p 通过靶向下调肺癌A549细胞中TM4SF1的表达抑制细胞的增殖、迁移、侵袭和裸鼠移植瘤生长。