ETHNOPHARMACOLOGICAL RELEVANCE:Paris polyphylla, a prominent traditional Chinese medicinal herb known for its heat-clearing and detoxifying properties, is widely utilized to relieve pain and treat sores, snake bites, and traumatic injuries. Recent pharmacological investigations have revealed its marked anti-tumor activity. However, as research continues to advance, its potential organ toxicity has been identified, presenting certain limitations for related studies. PURPOSE:This study aimed to compare the toxic components, toxic doses, and organ-specific toxic responses of the cultivated and wild varieties of two P. polyphylla cultivars, and to elucidate potential mechanisms underlying variety-specific hepatotoxicity. METHODS:Using ultra-performance liquid chromatography-tandem mass spectrometry, we identified the major active components in cultivated and wild varieties of P. polyphylla cultivars: P. polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz, referred to as PY, and P. polyphylla Smith var. chinensis (Franch.) Hara, referred to as PC. A comparative analysis was conducted to assess the differences in their contents. Additionally, we evaluated both the acute and subchronic toxicity of these varieties in mice to determine the differences in organ damage caused by the cultivated and wild varieties of PY and PC across various doses. This evaluation was based on the changes in mouse body weight, organ coefficients, and serum biochemical indicator levels, as well as histopathological alterations. Focusing on cultivated PY, we investigated its potential hepatotoxicity targets and the underlying mechanisms using a combination of network toxicology and transcriptomic techniques. The findings were validated through western blotting, terminal deoxynucleotidyl transferase dUTP nick end labeling, and transmission electron microscopy. RESULTS:Quantitative analysis revealed that PY contained higher levels of active components than PC, with wild varieties exceeding cultivated ones. Acute toxicity test revealed that the maximum tolerated doses of the extracts from both cultivated and wild PY and PC varieties exceeded 80 g/kg body weight in mice. In the subchronic toxicity test, when the doses of PY and PC exceeded 1.95 g/kg body weight, significant increases were observed in the levels of liver and heart function biomarkers. Pathological examinations primarily revealed inflammatory changes in the liver and heart in several cases. Notably, wild varieties elicited more pronounced organ damage than cultivated ones. Comprehensive network toxicology and transcriptomic analyzes revealed that P. polyphylla intervention disrupted hepatic metabolic homeostasis through pathways that impair mitochondrial structure and function, while simultaneously activating inflammatory response pathways and promoting hepatocyte apoptosis. Enrichment analysis revealed several key targets, including STAT3. Further experimental results demonstrated that cultivated PY significantly elevated inflammatory factor levels in mice. In liver tissue, there was an aberrant activation of apoptosis and mitochondrial autophagy, accompanied by abnormal expression of related molecules, such as STAT3, Bcl-2, and BNIP3. These research findings indicate that c-PY extract stimulates abnormal mitochondrial autophagy activation, induces apoptosis and inflammatory responses in liver tissue, and ultimately leads to liver damage, potentially associated with the activation of the STAT3/BNIP3 axis. Our study also highlights the efficacy and potential of integrating network toxicology and transcriptomic analyzes in toxicological studies. CONCLUSION:In this study, we compared the composition and toxic effects of cultivated and wild PY and PC varieties. Using transcriptomics and network toxicology, we revealed potential mechanisms of cultivated PY-induced hepatotoxicity. This research lays the foundation for determining the quality standards of P. polyphylla, evaluating the safety of its medicinal use, and elucidating the mechanisms underlying its toxicity.

BACKGROUND:Distant metastasis of colorectal cancer (CRC) is strongly driven by metabolic reprogramming and epithelial-mesenchymal transition (EMT). Increasing evidence suggests that these two processes form a reinforcing positive feedback loop; however, the integrated regulatory mechanism and its potential for pharmacological intervention remain insufficiently understood. OBJECTIVE:This study aimed to elucidate the mechanistic coupling between autophagy, metabolic reprogramming, and EMT, and to develop a targeted pharmacological strategy capable of disrupting this positive feedback loop. STUDY DESIGN:We systematically constructed and validated an autophagy-metabolism-phenotypic transformation regulatory axis centered on ATG4B and PKM2, and evaluated the therapeutic efficacy of Curcumol as a pathway-specific natural compound intervention. METHODS:Biochemical assays, protein-protein interaction analyses, and functional experiments were performed to determine how ATG4B regulates PKM2 Tyr105 phosphorylation, nuclear translocation, and glycolytic activity. Curcumol was applied to assess its ability to activate ATG4B-dependent autophagy and inhibit PKM2 activation. Anti-tumor efficacy was validated using colorectal cancer organoids, orthotopic implantation, and liver metastasis mouse models. RESULTS:ATG4B was identified as a core autophagy enzyme that directly binds to and shields the PKM2 Tyr105 site, preventing FGFR1-mediated phosphorylation and nuclear translocation. This blockade suppressed the Warburg effect, reduced lactate production, and synergistically inhibited EMT progression. Curcumol activated ATG4B-dependent autophagy, inhibited PKM2 activation, and effectively disrupted the metabolism-EMT positive feedback loop. In multiple CRC models, Curcumol markedly suppressed tumor growth and metastasis, supporting its therapeutic potential. CONCLUSION:This study reveals the ATG4B-PKM2 axis as a critical regulatory node linking autophagy, metabolic reprogramming, and EMT. Targeting this axis with Curcumol provides a precise strategy to interrupt metabolism-phenotype coupling, offering a mechanistically grounded and translationally promising approach for inhibiting CRC progression and metastasis.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) promotes the degradation of MHC-I molecules, thereby weakening CD8⁺ T cell–mediated immune surveillance. However, whether PCSK9 additionally regulates immune infiltration through the tumor vascular barrier remains unknown. This study employed a colorectal cancer (CRC) organoid–T cell coculture system, human umbilical vein endothelial cell (HUVEC) barrier assays, and a patient-derived orthotopic xenograft (PDOX) liver metastasis model. PCSK9 was knocked down to assess its effects on antigen presentation, T cell activation, endothelial integrity, and tumor growth. PCSK9 knockdown significantly upregulated MHC-I expression, enhanced CD8⁺ T cell activation and cytotoxicity, and induced high production of IFN-γ and TNF-α. These cytokines disrupted VE-cadherin–mediated endothelial junctions, reduced transendothelial electrical resistance (TEER), increased vascular permeability, and promoted deep T cell infiltration, ultimately suppressing tumor growth. Consistent in vitro and in vivo findings provided closed-loop validation from molecular to histological levels. For the first time, this study reveals that PCSK9 establishes a multilevel immunoregulatory axis spanning antigen presentation, T cell activation, and the vascular barrier, driving immune evasion and microenvironmental remodeling in CRC. Inhibition of PCSK9 simultaneously activates antitumor immunity and optimizes the vascular microenvironment, presenting a promising combinatorial therapeutic target against CRC metastasis.

OBJECTIVE:To evaluate the effects of Huangqi (Radix Astragali Mongolici)-Ezhu (Rhizoma Curcumae Phaeocaulis) (HQEZ) on colorectal cancer therapies and to elucidate the potential mechanisms of HQEZ, especially in combination with 5-Fluorouracil (5-FU). METHODS:The anti-tumor effects of HQEZ were evaluated in colorectal cancer models both in vivo and in vitro. The network pharmacological assay was used to investigate potential mechanisms of HQEZ. Potential target genes were selected by Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, protein-protein interaction network (PPI) and molecular docking. Within key targets, potential targets related to drug sensitivity, especially the sensitivity to 5-FU, were evaluated in HCT116 in vitro by immunofluorescence, quantitative real-time polymerase chain reaction (qPCR) and Western-blot. Then, changes in potential targets were assessed in tumors from tumor-bearing mice and the expression of these targets was also evaluated in colorectal cancer (COAD) patients from the Cancer Genome Atlas Program (TCGA) database. RESULTS:HQEZ significantly enhanced the anti-tumor activity of 5-FU in vivo and inhibit the growth of HCT116 in vitro. By network pharmacological analysis, key targets, such as protein kinase B (AKT1), epidermal growth factor receptor (EGFR), adenosine triphosphate (ATP) binding cassette subfamily B member 1 (ABCB1, also named multidrug resistance protein 1, MDR1), ATP binding cassette subfamily G member 2 (ABCG2), thymidylate synthetase (TYMS, also named TS), prostaglandin-endoperoxide synthase 2 (PTGS2), matrix metallopeptidase 2 (MMP2), MMP9, toll like receptor 4 (TLR4), TLR9 and dihydropyrimidine dehydrogenase (DPYD), were identified. Additionally, 4 potential core active ingredients (Folate, Curcumin, quercetin and kaempferol) were identified to be important for the treatment of colorectal cancer with HQEZ. In key targets, chemoresistance related targets were validated to be affected by HQEZ. Furthermore, 5-FU sensitivity related targets, including MDR1, TS, EGFR, ribonucleotide reductase catalytic subunit M1, Breast and Ovarian Cancer Susceptibility Protein 1 (BRCA1) and mutl homolog 1 were also significantly reduced by HQEZ both in vitro and in vivo. Finally, these validated key targets and 5-FU sensitivity related targets were demonstrated to be up-regulated in COAD patients based on TCGA database. CONCLUSION:HQEZ has synergistic effects on the anti-tumor activity of 5-FU in the treatment of colorectal cancer both in vivo and in vitro. The beneficial effect of HQEZ results from the inhibition of the drug sensitivity targets associated with 5-FU. The combination therapy of HQEZ with 5-FU or other chemotherapeutic drugs will also improve the anti-tumor efficacy of chemotherapy.

BACKGROUND:Colorectal cancer is one of the common malignant tumors in clinical practice, and traditional Chinese medicine, as an important adjuvant treatment method, plays important roles in the treatment of malignant tumors. OBJECTIVE:This study aims to explore the mechanism of action of the Qizhu anti-cancer recipe on colorectal cancer through transcriptome sequencing. METHODS:The control group and Qizhu anti-cancer recipe group were established separately, and sequencing of the cells of the two groups was performed using the Illumina sequencing platform. Two sets of Differentially Expressed Genes (DEGs) were screened using the DESeq2 algorithm, and Principal Component Analysis (PCA), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome, Disease Ontology (DO), and Protein-Protein Interaction (PPI) were used to comprehensively analyze the molecular functions and signaling pathways enriched by DEGs. RESULT:A total of 122 DEGs were identified through differential analysis, including 24 upregulated genes and 98 downregulated genes. GO analysis showed that DEGs were mainly enriched in functions such as alkaline phase activity, ion transport, cell differentiation, etc.; KEGG analysis showed that DEGs were mainly enriched in pathways such as Thiamine metabolism, apoptosis, signaling pathways regulating pluripotency of stem cells, cellular senescence and so on. Reactom analysis showed that DEGs were mainly enriched in response pathways such as EGR1,2,3 bind to the NAB2 promoter, EGR binds ARC gene, EGR-dependent NAB2 gene expression, etc.; DO analysis showed that differentially expressed genes were mainly enriched in diseases such as disease of cellular proliferation, disease of anatomical entity, organ system cancer, etc.; PPI analysis identified key differentially expressed genes, including DDIT3, CHAC1, TRIB3, and ASNS. CONCLUSION:Based on transcriptome sequencing and bioinformatics analysis, it was found that the Qizhu anti-cancer recipe may involve DEGs and signaling pathways in the treatment of colorectal cancer. Our study may provide potential drug targets for developing new treatment strategies for colorectal cancer.

Background: Owing to its remarkable efficacy, cisplatin (CDDP) is widely used as a chemotherapeutic drug in clinical cancer treatment; however, its severe nephrotoxicity often leads to acute kidney injury (AKI), in turn adversely affecting patient treatment and quality of life. Smilax glabra Roxb. (TFL), a Chinese herbal medicine, has various pharmacological effects, including antitumour, anti-inflammatory, and antioxidant activities, with the antioxidant activity being of useful in the detoxification of heavy metal toxicity. Aim: This study aimed to investigate, for the first time, the nephroprotective effects of TFL in alleviating CDDPinduced AKI and to elucidate its underlying mechanisms. Methods: In vitro and in vivo models of AKI were established using CDDP induction. For the in vivo model, CDDP (20 mg/kg) was intraperitoneally injected on day 7 to induce AKI. TFL treatment was administered daily at doses of 1.95 and 3.9 g/kg starting from the day 1 and continuing for 10 consecutive days. Blood samples were collected on day 10 after 72-h of CDDP injection for analysis. Kidney pathology was observed using haematoxylin and eosin (HE) staining, and mitochondrial ultrastructure was assessed using transmission electron microscopy. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), HO-1, NQO1, caspase-3, and cytochrome C (CYT-C) were determined using western blotting, PCR, and immunofluorescence (IF). Adenosine triphosphate (ATP) levels, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) were measured using the corresponding kits. Lastly, reverse validation of the Nrf2/HO-1 pathway was performed using the Nrf2-specific inhibitor, ML385. Results: After induction with 40 mu M CDDP, HK2 cells showed obvious mitochondrial damage, and the protein and mRNA expressions of Nrf2, HO-1, and NQO1 were inhibited, but gradually increased with TFL treatment. Furthermore, CDDP-induced AKI in mice was similar to the observations in the in vitro model using HK2 cells. The protective effects of TFL were reversed with ML385 therapy. Conclusion: In both in vivo and in vitro experiments, TFL activated the Nrf2/HO-1 signalling pathway, promoting the expression of antioxidant enzymes and thereby ameliorating CDDP-induced oxidative stress, mitochondrial dysfunction and renal cell apoptosis.

BACKGROUND:In colorectal cancer (CRC), aberrant tumor vasculature and epithelial-mesenchymal transition (EMT) synergistically drive distant metastasis, profoundly influencing patient prognosis. However, the underlying molecular mechanism and effective systemic therapeutic strategies remain unclear. PURPOSE:This study aimed to identify key molecular drivers of the vascular anomaly-EMT-metastasis axis in CRC and to evaluate the therapeutic potential of Curdione, a bioactive natural compound, in disrupting this pathological loop. STUDY DESIGN:We employed integrated in vitro and in vivo experimental systems, supported by multi-omics and single-cell analyses, to investigate the role of phospholipase C gamma 1 (PLCγ1) in CRC progression and its modulation by Curdione. METHODS:CRC patient-derived organoid and animal metastasis models were established, and functional assays were conducted to assess tumor invasiveness, vascular integrity, and EMT phenotype. Multi-omics profiling and mechanistic studies identified molecular targets and signaling pathways. Combination therapy with Curdione and Bevacizumab was further evaluated to determine synergistic effects. RESULTS:Multi-omics analyses identified PLCγ1 as a central target of Curdione. Mechanistic analyses suggest that PLCγ1 is involved in the regulation of vascular abnormality, endothelial barrier dysfunction, and EMT, at least in part through the PKC-PI3K signaling pathway, which is associated with features of a prometastatic microenvironment. Curdione treatment was associated with enhanced PLCγ1 degradation and attenuation of downstream signaling, accompanied by improved vascular integrity, reinforced epithelial stability, and a reduction in metastatic phenotypes Moreover, combined treatment with Curdione and Bevacizumab was associated with enhanced anti-angiogenic effects and reduced EMT-associated features. CONCLUSION:This study identifies the PLCγ1-PKC-PI3K signaling axis as a key pathway associated with the coordination of vascular abnormality, EMT, and metastatic phenotypes in CRC. Our findings suggest that Curdione functions as a natural multi-target modulator of the tumor microenvironment and, when combined with Bevacizumab, may enhance anti-metastatic efficacy in CRC models.

Background: Abnormal tumor blood vessels can significantly promote the malignant progression of tumors, prompting researchers to focus on drugs that normalize these vessels for clinical treatment. The combination of the Qi-tonifying drug Astragali Radix and the blood-activating drug Curcumae Rhizoma, referred to as AC, exhibited significant anti-tumor metastasis effects. However, the association between the anti-tumor metastasis effect of AC and its potential role in regulating tumor vascular remodeling warrants further exploration. Purpose: This study aimed to elucidate the mechanism through which AC induces tumor blood vessel normalization in colon cancer (CC). Methods: The potential active components of AC were identified through UPLC-MS/MS. An orthotopic transplantation model of CC was established in BALB/c mice using the CT26-Lucifer cell line, and the effects of AC were evaluated using IVIS imaging, hematoxylin and eosin (H&E) staining, and immunohistochemistry. Network pharmacology and molecular biology analyses were employed to identify the potential direct targets of AC. Subsequently, RT-PCR and Western blotting techniques were utilized to validate the findings obtained from network pharmacology. Furthermore, ELISA and other methodologies were used to investigate glycolysis-related indicators, along with immunofluorescence technology to demonstrate changes in vascular leakage and perfusion characteristics associated with blood vessel normalization. Results: We identified HIF-1 alpha as a potential direct target of AC. This interaction influences the glycolytic processes in both tumor cells and tumor-associated endothelial cells (TECs) by directly binding to HIF-1 alpha and modulating its nuclear translocation, thereby determining the integrity of TEC junctions. Mechanistically, AC directly regulates the key enzyme PFKFB3 in glycolysis by modulating HIF-1 alpha expression and inhibiting its nuclear translocation. This action reduces tumor glycolytic flux, decreases the internalization of VE-cad, and influences the expression of downstream matrix metalloproteinases (MMPs), thereby strengthening the adherens and tight junctions between TECs and restoring vascular integrity. Conclusion: This study presents novel findings that AC can regulate glycolysis through the inhibition of HIF-1 alpha nuclear translocation, thereby promoting the normalization of tumor blood vessels and effectively inhibiting tumor metastasis. These results suggested that AC may serve as an effective therapeutic agent for normalizing tumor blood vessels.

OBJECTIVE: To evaluate the effects of Huangqi(Radix Astragali Mongolici)-Ezhu(Rhizoma Curcumae Phaeocaulis)(HQEZ) on colorectal cancer therapies and to elucidate the potential mechanisms of HQEZ, especially in combination with 5-Fluorouracil(5-FU). METHODS: The anti-tumor effects of HQEZ were evaluated in colorectal cancer models both in vivo and in vitro. The network pharmacological assay was used to investigate potential mechanisms of HQEZ. Potential target genes were selected by Gene Ontology(GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis, protein-protein interaction network(PPI) and molecular docking. Within key targets, potential targets related to drug sensitivity, especially the sensitivity to 5-FU, were evaluated in HCT116 in vitro by immunofluorescence, quantitative real-time polymerase chain reaction(qPCR) and Western-blot. Then, changes in potential targets were assessed in tumors from tumor-bearing mice and the expression of these targets was also evaluated in colorectal cancer(COAD) patients from the Cancer Genome Atlas Program(TCGA) database. RESULTS: HQEZ significantly enhanced the anti-tumor activity of 5-FU in vivo and inhibit the growth of HCT116 in vitro. By network pharmacological analysis, key targets, such as protein kinase B(AKT1), epidermal growth factor receptor(EGFR), adenosine triphosphate(ATP) binding cassette subfamily B member 1(ABCB1, also named multidrug resistance protein 1, MDR1), ATP binding cassette subfamily G member 2(ABCG2), thymidylate synthetase(TYMS, also named TS), prostaglandinendoperoxide synthase 2(PTGS2), matrix metallopeptidase 2(MMP2), MMP9, toll like receptor 4(TLR4), TLR9 and dihydropyrimidine dehydrogenase(DPYD), were identified. Additionally, 4 potential core active ingredients(Folate, Curcumin, quercetin and kaempferol) were identified to be important for the treatment of colorectal cancer with HQEZ. In key targets, chemoresistance related targets were validated to be affected by HQEZ. Furthermore, 5-FU sensitivity related targets, including MDR1, TS, EGFR, ribonucleotide reductase catalytic subunit M1, Breast and Ovarian Cancer Susceptibility Protein 1(BRCA1) and mutl homolog 1 were also significantly reduced by HQEZ both in vitro and in vivo. Finally, these validated key targets and 5-FU sensitivity related targets were demonstrated to be up-regulated in COAD patients based on TCGA database. CONCLUSION: HQEZ has synergistic effects on the antitumor activity of 5-FU in the treatment of colorectal cancer both in vivo and in vitro. The beneficial effect of HQEZ results from the inhibition of the drug sensitivity targets associated with 5-FU. The combination therapy of HQEZ with 5-FU or other chemotherapeutic drugs will also improve the anti-tumor efficacy of chemotherapy.

Background: Hypoxia-induced glycolysis represents a hallmark of colorectal cancer (CRC) progression and contributes significantly to therapeutic resistance. Curcumol, a natural sesquiterpenoid derived from Curcumae Rhizoma, has demonstrated promising anti-tumor properties. However, its impact on metabolic reprogramming under hypoxic conditions remains largely undefined. Objective: The objective of this study was to elucidate the potential of Curcumol in inhibiting glycolytic reprogramming and impede CRC progression via regulation of the VHL/HIF-1α signaling pathway. Methods: CRC cells and orthotopic mouse models were treated with Curcumol under chemically induced hypoxic conditions. Metabolic alterations were evaluated using Seahorse extracellular flux analysis, Western blot analysis, quantitative real-time PCR (qRT-PCR), immunohistochemistry (IHC) and co-immunoprecipitation (Co-IP). Functional validation of glycolysis and epithelial–mesenchymal transition (EMT) phenotypes was conducted through in vitro and in vivo assays. Results: Curcumol inhibited HIF-1α-mediated metabolic reprogramming by upregulating VHL expression, thereby promoting HIF-1α degradation. This effect led to the downregulation of key glycolytic genes (HK2, LDHA, and GLUT1), decreased glycolytic flux, and lactate production, ultimately suppressing CRC cell proliferation and invasion. The anti-tumor efficacy of Curcumol was validated in both in vitro and in vivo models. Moreover, Curcumol effectively reversed the hypoxia-induced epithelial–mesenchymal transition (EMT) phenotype, suggesting that its metabolic regulatory effects may contribute to reduced metastatic potential. Conclusions: Curcumol suppresses glycolysis and CRC progression by activating the VHL/HIF-1α signaling axis. These findings underscore the potential of Curcumol as a natural metabolic regulator capable of reversing tumor metabolic reprogramming, offering a promising therapeutic strategy for CRC treatment.

Ethnopharmacological relevance: Bupleurum chinense DC.-Scutellaria baicalensis Georgi (BS) is a classic drug pair that has good clinical effects on depression and many tumors. However, the concurrent targeting mechanism of how the aforementioned drug pair is valid in the two distinct diseases, has not been clarified yet. Aim of the study: The components of BS were detected by LC-MS, combined with network pharmacology to explore the active ingredients and common targeting mechanism of its multi-pathway regulation of BS in treating depression and CRC, and to validate the dual effects of BS using the CUMS mice model and orthotopic transplantation tumor mice model of CRC. Results: Twenty-nine components were screened, 84 common gene targets were obteined, and the top 5 key targets including STAT3, PIK3R1, PIK3CA, AKT1, IL-6 were identified by PPI network. GO and KEGG analyses revealed that PI3K/AKT and JAK/STAT signaling pathways might play a crucial role of BS in regulating depression and CRC. BS significantly modulated CUMS-induced depressive-like behavior, attenuated neuronal damage, and reduced serum EPI and NE levels in CUMS model mice. BS improved the pathological histological changes of solid tumors and liver tissues and inhibited solid tumors and liver metastases in tumor-bearing mice. BS significantly decreased the proteins' expression of IL-6, p-JAK2, p-STAT3, p-PI3K, p-AKT1 in hippocampal tissues and solid tumors, and regulated the levels of IL-2, IL-6 and IL-10 in serum of two models of mice. Conclusion: BS can exert dual antidepressant and anti-CRC effects by inhibiting the expression of IL-6/JAK2/ STAT3 and PI3K/AKT pathway proteins and regulating the release of inflammatory cytokines.

Pyroptosis is a type of programmed cell death mediated by gasdermines (GSDMs). The N-terminal domain of GSDMs forms pores in the plasma membrane, causing cell membrane rupture and the release of cell contents, leading to an inflammatory response and mediating pyrodeath. Pyroptosis plays an important role in inflammatory diseases and malignant tumors. With the further study of pyroptosis, an increasing number of studies have shown that the pyroptosis pathway can regulate the tumor microenvironment and antitumor immunity of colorectal cancer and is closely related to the occurrence, development, treatment and prognosis of colorectal cancer. This review aimed to explore the molecular mechanism of pyroptosis and the role of pyroptosis in the occurrence, development, treatment and prognosis of colorectal cancer (CRC) and to provide ideas for the clinical diagnosis and treatment of CRC.

BackgroundColorectal cancer (CRC) is one of the most common causes of cancer-related mortality and significantly impairs quality of life. Astragali Radix-Curcumae Rhizoma (AC) is widely employed in the treatment of CRC in Chinese medicine, but the precise mechanisms remain unclear.PurposeThis study aimed to elucidate the mechanisms by which AC inhibits CRC progression.MethodsThe active components of AC were identified using UPLC-MS/MS analysis. An orthotopic transplantation colorectal tumor model was established in BALB/c mice using the CT26-Lucifer cell line to evaluate the effects of AC. Tumor volumes were monitored using IVIS imaging technology. Histological examination of tumor morphology was performed with hematoxylin and eosin (H&E) staining. Transcriptomic sequencing of mouse tumor samples was conducted to identify critical pathways and molecular targets. The impact of AC on cell viability and migration was assessed using CCK-8 and wound healing assays, respectively. To investigate the effects of AC on CRC cells, an in vitro hypoxic model was established using cobalt chloride (CoCl2), a hypoxia inducer. HIF-2α overexpression was achieved by constructing stable lentiviral vectors. Key targets identified from RNA-seq, such as c-Myc, Ki-67, β-catenin, cleaved caspase 3, CD133, and CD44, were evaluated using western blotting, qRT-PCR, and immunofluorescence assays. Epithelial-Mesenchymal Transition (EMT) and spheroid cloning assays were employed to evaluate phenotypic changes in cancer stem cells.ResultsTwelve components of AC were identified. AC effectively inhibited CRC progression in vivo. Transcriptomic analysis highlighted hypoxic signaling as a significantly enriched pathway, implicating its role in suppressing CRC progression by AC. In the hypoxic model, AC inhibited the proliferation and migration of CRC cells in vitro. Furthermore, AC reduced cancer stemness by downregulating stemness markers, inhibiting EMT, and decreasing tumor sphere formation. The downregulation of hypoxic responses and the shift in stemness by AC involved attenuation of HIF-2α and WNT/β-catenin signaling.ConclusionThis study provides the first evidence that AC reduces the stemness of CRC and the inhibition of the transition of CRC to stem-like cells by AC is closely related to the downregulation of the HIF-2α/β-catenin pathway, especially under hypoxic conditions.

The animal and cell models were used in this study to investigate the mechanism of Astragali Radix-Curcumae Rhizoma(HQEZ) in inhibiting colon cancer progression and enhancing the efficacy of 5-fluorouracil(5-FU) by regulating hypoxia-inducible factors and tumor stem cells. The animal model was established by subcutaneous transplantation of colon cancer HCT116 cells in nude mice, and 24 successfully modeled mice were randomized into model, 5-FU, HQEZ, and 5-FU+HQEZ groups. The tumor volume was measured every two days. Western blot was employed to measure the protein levels of epidermal growth factor receptor(EGFR), dihydropyrimidine dehydrogenase(DPYD), and thymidylate synthase(TYMS), the key targets of the hypoxic core region, as well as the hypoxia-inducible factors HIF-1α and HIF-2α and the cancer stem cell surface marker CD133 and SRY-box transcription factor 2(SOX2). The results of animal experiments showed that HQEZ slowed down the tumor growth and significantly increased the tumor inhibition rate of 5-FU. Compared with the model group, HQEZ significantly down-regulated the protein levels of EGFR and DPYD, and 5-FU+HQEZ significantly down-regulated the protein levels of EGFR and TYMS in tumors. Compared with the model group, HQEZ significantly down-regulated the protein levels of HIF-1α, HIF-2α, SOX2, and CD133 in the hypoxic core region. Compared with the 5-FU group, 5-FU+HQEZ lowered the protein levels of HIF-1α, HIF-2α, and SOX2. The cell experiments showed that the protein le-vels of HIF-1α and HIF-2α in HCT116 cells elevated significantly after low oxygen treatment. Compared with 5-FU(1.38 μmol·L~(-1)) alone, HQEZ(40 mg·mL~(-1)) and 5-FU+HQEZ significantly down-regulated the protein levels of HIF-1α, HIF-2α, and TYMS. In conclusion, HQEZ can inhibit the expression of hypoxia-responsive molecules in colon cancer cells and reduce the properties of cancer stem cells, thereby enhancing the therapeutic effect of 5-FU on colon cancer.

Background Colorectal cancer (CRC) progression is strongly influenced by metabolic reprogramming, particularly under hypoxic conditions, where hypoxia-inducible factor 1 alpha (HIF-1α) acts as a critical driver. HIF-1α upregulates glycolytic enzymes, promoting both tumor cell survival and invasive potential. Excessive lactate production, resulting from heightened glycolysis, not only facilitates tumor adaptation to hypoxia but also acidifies the tumor microenvironment, leading to immune suppression. Targeting HIF-1α and lactate metabolism has therefore emerged as a promising therapeutic approach in CRC. However, most existing therapies focus on inhibiting HIF-1α expression without addressing its degradation pathways. Curcumol, a sesquiterpenoid derived from Curcuma species, exhibits a unique mechanism by promoting HIF-1α degradation via the von Hippel-Lindau (VHL) pathway. This dual action of curcumol—restoring VHL activity while suppressing HIF-1α stability—offers a novel approach for interfering with CRC metabolic reprogramming. Methods This study employed two approaches to validate the therapeutic effect of curcumol, including an in vivo Balb/c mouse orthotopic tumor transplantation model and an in vitro hypoxia model of colorectal cancer cell lines. Hematoxylin and eosin (HE) staining was performed to examine pathological changes in tumor tissues, and immunohistochemistry was used to verify the positive expression rate of HIF-1α in tumor tissues. The XF 96 Seahorse metabolic analyzer was utilized to measure the glycolytic metabolism pathway in colorectal cancer cell lines. In addition, Western blotting, immunofluorescence, and q-PCR were applied to assess the expression levels of HIF-1α and glycolysis-related markers in tumor tissues and colorectal cancer cells. Glycolytic enzyme activity assay kits were also used to detect the activity of key glycolytic enzymes, while co-immunoprecipitation (Co-IP) was conducted to evaluate the interaction between HIF-1α and VHL. Furthermore, a VHL gene knockout mouse model was established to investigate curcumol's effect on the VHL-HIF-1α pathway. Results Curcumol significantly restored VHL expression, reduced HIF-1α stability, and downregulated glycolytic genes (HK2, LDHA), leading to reduction in lactate production. Moreover, curcumol improved the acidic conditions of the tumor microenvironment by lowering lactate levels, and suppressing CRC cell proliferation and invasion. Conclusion Curcumol inhibits HIF-1α-mediated metabolic reprogramming, significantly improving the CRC tumor microenvironment. Future studies should validate curcumol’s efficacy in animal models and explore its potential for combination with metabolic inhibitors or chemotherapy, offering a promising therapeutic strategy for advanced CRC.

IntroductionAlterations in the gut microbiome and bile acid metabolism are known to play a role in the development and progression of colon cancer. Medicinal plants like Astragalus mongholicus Bunge and Curcuma aromatica Salisb. (AC) have shown preferable therapeutic effect on cancer therapy, especially digestive tract tumors like colon cancer. However, the precise mechanisms of AC inhibiting colon cancer, particularly in relation to the gut microbiome and bile acid dynamics, are not fully understood.MethodsOur research aimed to investigate the anti-tumor properties of AC in mice with CT26 colon cancer and further investigate its underlying mechanism via intestinal microbiota. The size and pathological changes of solid tumors in colon cancer are used to evaluate the inhibitory effect of AC on colon cancer. Metagenomics and 16s rRNA gene sequencing were employed to clarify the dysbiosis in the gut microbiome of colon cancer and its impact on colon cancer. The levels of bile acids (BAs) in the feces of mice from each group were measured using UPLC-Qtrap-MS/MS.ResultsAC effectively suppressed the growth of colon cancer and reduced histological damage. Notably, AC treatment led to changes in the gut microbiome composition, with a decrease in pathogenic species like Citrobacter and Candidatus_Arthromitus, and an increase in beneficial microbial populations including Adlercreutzia, Lachnospiraceae_UCG-001, and Parvibacter. Additionally, AC altered bile acid profiles, resulting in a significant decrease in pro-carcinogenic bile acids such as deoxycholic acid (DCA) and lithocholic acid (LCA), while increasing the concentration of the cancer-inhibitory bile acid, ursodeoxycholic acid (UDCA). Tracking and analyzing the data, AC may mainly upregulate FabG and baiA genes by increasing the relative abundance of Adlercreutzia and Parvibacter bacteria, which promoting the metabolism of pro-carcinogenic LCA.DiscussionThese findings provide strong evidence supporting the role of AC in regulating gut microbiome-mediated bile acid metabolism, which is crucial in impeding the progression of colon cancer.

Objective: To evaluate the effect of Sancao Lichang decoction as traditional Chinese medicine on diarrhea-predominant irritable bowel syndrome (IBS-D) and TLR4/MyD88/NF-κB pathway. Background: Traditional Chinese medicine has made significant progress in preventing and treating irritable bowel syndrome, and its efficacy has been validated by clinical practice. Sancao Lichang decoction is an empirical prescription developed by professor Tang Decai that has been used for many years to treat chronic diarrhoea with good curative effec. Still, its mechanism of action on IBS-D is unknown. Methods: The study sample of Fifty SD rats was randomly divided into a blank group, model group, low-dose group, medium-dose group, and high-dose group (n = 10). The IBS-D rat models were established by restraining stress method and acetic acid enema. After different treatments, defecation frequency, fecal water content (FWC), serum IL-6 and TNF-α contents, and protein level of TLR4/MyD88/NF-κB in colon tissues were detected separately. Results: The indexes of rats in each group were significantly different. The increase in body weight in the medium-dose and high-dose groups was significantly higher than that in the model group (p < 0.05). Compared with the model group, the medium and high dose groups had lower diarrhea frequency, FWC, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) (p < 0.05). The expression levels of TLR4, MyD88, and NF-κB protein in the colon of the three groups treated with Sancao-Lichan decoction were significantly lower than those in the model group (p < 0.01). After different treatments, the colonic mucosa of rats in each group was stained with HE, which proved that the structural damage of colonic mucosa was improved after treatment with Sancao Lichang decoction, and the improvement effect was dose-dependent. Conclusion: Sancao Lichang decoction may reduce IBS-D by inhibiting TLR4/MyD88/NF-κB pathway, inhibiting the inflammatory response, and improving intestinal mucosal barrier function.

Traditional Chinese drug effects are a highly generalized function of drug treatment and health care under the guidance of the theory of Chinese medicine. The identification of each drug effect has experienced a long historical evolution. Throughout the records and applications in ancient and modern herbal literature， there are different degrees of expression， which may be attributed to the differences in geographical space， the understanding of doctors， the inheritance of academic schools， and the selection of compatible environments. The redefinition of traditional Chinese drug effects merits an in-depth discussion. Clarifying the "source" and "flow" of the drug effects， considering the combination of basic effects and prescription， and evaluating the dosage of drugs should be the main principles of the research. On the basis of inheritance and innovation， it is proposed that we attach importance to the collation of traditional Chinese drug effects in ancient and modern literature， strengthen the inheritance of the clinical medication experiences of famous historical Chinese medicines， introduce modern clinical and pharmacological research， and use the phylogenetic theory of medicinal plants as the main approaches to study traditional Chinese drug effects. This has far-reaching significance for unifying and standardizing traditional Chinese drug effects， improving clinical efficacy， and broadening the clinical application scope of traditional Chinese drugs.

This study aims to investigate the effect of Astragali Radix-Curcumae Rhizoma(AC) combination on the proliferation, migration, and invasion of colon cancer HT-29 cells based on epithelial-mesenchymal transition(EMT). HT-29 cells were respectively treated with 0, 3, 6 and 12 g·kg~(-1) AC-containing serum for 48 h. The survival and growth of cells were measured by thiazole blue(MTT) colorimetry, and the proliferation, migration, and invasion of cells were detected by 5-ethynyl-2'-deoxyuridine(EdU) test and Transwell assay. Cell apoptosis was examined by flow cytometry. The BALB/c nude mouse model of subcutaneous colon cancer xenograft was established, and then model mice were classified into blank control group, 6 g·kg~(-1) AC group, and 12 g·kg~(-1) AC group. The tumor weight and volume of mice were recorded, and the histopathological morphology of the tumor was observed based on hematoxylin-eosin(HE) staining. The expression of apoptosis-associated proteins B-cell lymphoma-2-associated X protein(Bax), cysteine-aspartic acid protease-3(caspase-3), and cleaved caspase-3, and EMT-associated proteins E-cadherin, MMP9, MMP2 and vimentin in HT-29 cells and mouse tumor tissues after the treatment of AC was determined by Western blot. The results showed that cell survival rate and the number of cells at proliferation stage decreased compared with those in the blank control group. The number of migrating and invading cells reduced and the number of apoptotic cells increased in the administration groups compared with those in the blank control group. As for the in vivo experiment, compared with the blank control group, the administration groups had small tumors with low mass and shrinkage of cells and karyopycnosis in the tumor tissue, indicating that the AC combination may improve EMT. In addition, the expression of Bcl2 and E-cadherin increased and the expression of Bax, caspase-3, cleaved caspase-3, MMP9, MMP2, and vimentin decreased in HT-29 cells and tumor tissues in each administration group. In summary, the AC combination can significantly inhibit the proliferation, invasion, migration, and EMT of HT-29 cells in vivo and in vitro and promote the apoptosis of colon cancer cells.