Published online Nov 15, 2026. doi: 10.4251/wjgo.121010
Revised: June 19, 2026
Accepted: August 14, 2026
Published online: November 15, 2026
Processing time: 194 Days and 17.4 Hours
Chronic constipation-associated inflammation is a global public health threat as end-stage colorectal malignancy continues to rise worldwide. Aberrant innate immune signaling, especially via NOD-like receptor protein 3 (NLRP3) inflammasome–caspase-1-interleukin-1 beta (IL-1β) cascade, has emerged as a molecular driver of chronic colonic epithelial inflammation in response to prolonged ex
To investigate whether ASP inhibits the NLRP3/caspase-1/IL-1β inflammasome signaling pathway to attenuate deoxycholic acid (DCA)-induced chronic colonic epithelial inflammation and early neoplastic transformation, thereby establishing a mechanistic basis for ASP as a chemopreventive candidate against constipation-associated colorectal cancer.
NCM460 cells – a well-characterized normal human colon mucosal epithelial cell line, were exposed separately to vehicle control, or used as osmotic control and treated with vehicle plus equimolar sodium chloride, or 200 μM DCA and co-treated with ASP at 2 increasing concentrations (ASP-L and ASP-H). Nigericin, a classical NLRP3 activator that was used to reinvite inflammasome activity, and a series of selective pharmacological probes – the NLRP3-specific inhibitors MCC950 and cycplase 09, the potassium ionophore nigericin as well as the caspase-1 selective inhibitor VX-765 – were utilized for mechanistic dissection. Immunofluorescence was used to determine the localization and abundance of NLRP3, ASC, and cleaved caspase-1. Reverse-transcription PCR and ELISA were used to measure messenger RNA and secreted protein levels of IL-1β, IL-18, tumor necrosis factor-alpha, IL-6, COX-2 and matrix metalloproteinase-9. Markers of epithelial-mesenchymal transition and cellular proliferation were further examined to determine early neoplastic transformation.
DCA challenge remarkably induced the expression of NLRP3, ASC and cleaved caspase-1 as well as all six downstream inflammatory mediators at both transcript and protein levels in NCM460 cells, along with up-regulating epithelial-mesenchymal transition marker genes and promoting cell proliferation. The changes to these parameters were dose-dependently reversed by ASP; in the case of the third exposure (higher concentration), all parameters were close to baseline levels. The activation of caspase-1 and IL-1beta by Nigericin was restored in the presence of ASP, but co-treating with either MCC950 or VX-765 completely inhibited this rebound proven that NLRP3/caspase-1 pathwayspecifiselectivity. Cycplase 09 and VX-765 significantly suppressed all six inflammatory mediators in ASP + nigericin cultures, as confirmed by reverse-transcription PCR and ELISA analyses.
ASP inhibits the NLRP3/caspase-1/IL-1β pathway and protects DCA-induced inflammatory injury of colonic epithelial cells and early neoplastic transformation in vitro. These mechanistic insights warrant additional preclinical and clinical testing of ASP as a chemopreventive agents for constipation-related colorectal cancer risk.
Core Tip: Chronic constipation has lately been considered a possible risk factor for colorectal cancer due to prolonged colonic transit time and retention of secondary bile acids, e.g., deoxycholic acid, which can lead to persistent epithelial inflammation. Here, we present that a bioactive component of the traditional Chinese medicine Yang-Gui [Angelica sinensis polysaccharide (ASP)] can effectively suppress deoxycholic acid-induced inflammatory processes in human colonic epithelial cells. Mechanistically, ASP is able to inhibit activation of the NOD-like receptor protein 3 inflammasome and its downstream cascades caspase-1/interleukin-1β signaling pathway, thereby reducing the production of several pro-inflammatory mediators in early colorectal carcinogenesis. Our findings give mechanistic support for ASP as a potential natural candidate for preventing inflammation-driven colonic carcinogenesis under chronic constipated condition.
- Citation: Huang X, Liu WCN, Li DX. Angelica sinensis polysaccharide attenuates constipation-associated colorectal carcinogenesis by inhibiting the NLRP3/Caspase-1/IL-1β inflammasome axis. World J Gastrointest Oncol 2026; 18(11): 121010
- URL: https://www.wjgnet.com/1948-5204/full/v18/i11/121010.htm
- DOI: https://dx.doi.org/10.4251/wjgo.121010
Over the last 30 years, the global prevalence of chronic functional constipation has increased significantly in recent decades and currently affects about 10%-15% of adults worldwide[1], along with a mounting body of epidemiological evidence indicating that prolonged colonic transit time increases the risk for colorectal cancer (CRC)[2]. CRC is one of the deadliest malignancies globally and follows a well-established adenoma–carcinoma sequence that unfolds over years to decades, providing an opportunity for early chemopreventive intervention[3,4]. Older estimates indicate that chronic constipation approx. doubles the risk of CRC among the general population[2], implying that colonisation-associated chronic inflammation represents an underappreciated epidemiological concern in terms of colorectal carcinogenesis. While pharmacologic approaches focused on laxative therapy and diet-based fiber supplementation have led to partial symptom control, a large demographic of patients sustain subclinical colonic inflammation with significant aspects remaining an unmet need for mechanistically distinct chemoprevention.
In the last decade, a paradigm-shift has redefined chronic sterile inflammation, as an integral pathophysiological mechanism of constipation-derived CRC inter-relation, augmenting traditional genetic and diet-driven paradigms[5]. The underlying pathophysiological mechanism revolves around the extended dwelling of luminal contents within the colon as a result of constipation, favoring bacterial metabolism of primary to secondary bile acids, particularly deoxycholic acid (DCA)[6]. Because colonic epithelial cells occupy this unique anatomical niche, where they are exposed to elevated, luminal concentrations of DCA during constipation[7], they represent some of the first cellular targets subjected to bile acid-mediated cytotoxicity. When overwhelmed with DCA, these cells liberate a wide range of proinflammatory cytokines, matrix-degrading enzymes and cyclooxygenase metabolites that together orchestrate immune cell recruitment, mucosal barrier disruption, oxidative DNA damage and the progressive epithelial-mesenchymal transition that is characteristic of early colorectal neoplasia.
NOD-like receptor protein 3 (NLRP3) inflammasome at the molecular level is of great research interest as a central integrator of danger signals in DCA-induced colonic inflammation. NLRP3 is a multiprotein cytoplasmic complex composed of the sensor NLRP3, the adaptor apoptosis-associated speck-like protein containing a CARD (ASC; also known as PYCARD), and the effector protease pro-caspase-1[8-10]. In the canonical pathway, diverse activators – such as reactive oxygen species (ROS), potassium efflux, lysosomal destabilization, and mitochondrial damage[8] – induce NLRP3 oligomerization and recruitment of ASC that polymerizes into large perinuclear “specks”, functioning to promote pro-caspase-1 proximity-induced autocleavage. The resulting active caspase-1 (p20/p10 tetramer) then proteolytically cleaves the inactive precursors pro-interleukin (IL)-1β and pro-IL-18 to yield their mature, bioactive forms, and at the same time cleaves gasdermin D to release N-terminal pore-forming fragments that enable pyroptotic cell death and unconventional cytokine secretion[11,12]. Once released to the extracellular milieu, IL-1β and IL-18 propagate localized inflammation in part through activating their cognate receptors expressed on neighboring epithelial cells, resident macrophages and inflammatory leukocyte infiltrates to create a self-perpetuating loop of inflammation driving pro
Natural products have long provided a fertile ground to identify lead compounds in the management of many inflammatory diseases and for cancer chemoprevention. Angelica sinensis (Oliv.) Diels, called Danggui in Chinese pharmacopoeia and one of the most commonly prescribed herbs for traditional Chinese medicine, is valued for its blood tonifying, analgesic and intestinal regulating properties[15]. Angelica sinensis polysaccharide (ASP), the water-soluble macromolecular fraction of this herb, exhibits a variety of immunomodulatory, anti-inflammatory, antioxidant and antitumor bioactivities in various experimental disease models[16-18]. Moreover, accumulated clinical experience has acknowledged that formulae based on Danggui have been used to relieve constipation and enhance the motility of intestines for a long time[19,20]. Although these advances have been made, the ability of ASP to intercept NLRP3/caspase-1/IL-1β-dependent inflammatory signaling in the particular context of DCA-induced colonic epithelial damage and early neoplastic transformation has not been rigorously explored.
We hypothesized that ASP suppresses DCA-induced colonic epithelial inflammation and early neoplastic trans
ASP [high performance liquid chromatography purity ≥ 98% (supplier)] DCA, sodium chloride, LPS from Escherichia coli serotype O111:B4, nigericin sodium salt and adenosine 5′-triphosphate disodium salt (ATP) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Four inflammasome modulating compounds were obtained from InvivoGen (San Diego, CA, United States) and Selleck Chemicals (Houston, TX, United States): (1) The selective NLRP3 inhibitor MCC950 (also referred to as CP-456773); (2) The direct NLRP3 ATPase-site inhibitor cycplase 09 (CY-09); (3) The potassium ionophore nigericin (to serve as a canonical NLRP3 activation signal); and (4) Caspase-1 selective inhibitor VX-765 (Belnacasan). Rabbit polyclonal antibodies against NLRP3, ASC/TMS1, cleaved caspase-1 (p20), CK20 and IL-1β were purchased from Abcam (Cambridge, United Kingdom) and Cell Signaling Technology (Danvers, MA, United States). Alexa Fluor 488-conjugated and 594-conjugated goat anti-rabbit IgG secondary antibodies (Thermo Fisher Scientific, Waltham, MA, United States), and 4′,6-diamidino-2-phenylindole (DAPI) was used as a nuclear counterstain. The Quantikine ELISA for IL-1β, IL-18, TNF-α, IL-6, COX-2 and matrix metalloproteinase-9 (MMP-9) were sourced from R&D Systems (Minneapolis, MN, United States). RNA isolation reagent (TRIzol), the PrimeScript reverse transcription kit, and TB Green Premix Ex Taq qPCR master mix were obtained from Takara Bio (Shiga, Japan).
The normal human colon mucosal epithelial cell line NCM460 was purchased from the INCELL Corporation (San Antonio, TX, United States). Cells were maintained and propagated routinely in RPMI-1640 medium supplemented with 10% heat-inactivated FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin in standard incubator conditions (37 °C, 5% CO2 saturated humidity). Downstream assays were only performed with passages 5-15.
Components of the dose-response arm were characterized by six different treatment arms: (1) Control: Culture in vehicle (0.1% DMSO); (2) OC: Culture in vehicle adding equimolar sodium chloride to control for osmolality; (3) DCA: Culture with DCA at 200 μM; (4) ASP-L: 200 μM DCA together with 100 μg/mL ASP; (5) ASP-H: Treatment with 200 μM DCA with (ASP) = 200 μg/mL; and (6) Nigericin: 200 μM DCA co-treated with 200 μg/mL ASP (ASP-H dose) plus nigericin (10 μM) administered for the final 1 hour of incubation following LPS priming (100 ng/mL, 4 hours), to pharmacologically reactivate NLRP3 inflammasome activity and confirm the pathway specificity of ASP-mediated inhibition. In the priming-activation protocol, cells were primed 4 hours with 100 ng/mL LPS before nigericin treatment to ensure sufficient pro-IL-1β and NLRP3 expression. All treatments were for 48 hours before harvest, unless otherwise indicated.
The second experiment was set up as a pathway validation and included ASP with individual or combined pharmacological agents: ASP + nigercin, ASP + MCC950, ASP + ATP, ASP + VX-765 and also the latter three agents in combination with either CY-09 or nigericin for stimulation of ASC. Preliminary optimization experiments had identified working concentrations for each compound.
NCM460 cells were seeded on sterile glass coverslips placed in 24-well culture plates and treated with the above-mentioned experiments. At the end of treatment, monolayers were washed with phosphate-buffered saline (PBS) and fixed in 4% paraformaldehyde (15 minutes, room temperature). Membrane permeabilization was then performed using 0.3% Triton X-100 (10 minutes) followed by a step of blocking for 1 hour with 5% bovine serum albumin. The coverslips were then incubated overnight at 4 °C with primary antibodies targeting NLRP3, ASC, cleaved caspase-1 (p20) or IL-1β (each at a dilution of 1:200), and co-stained for anti-CK20 (1:100) to mark colonic epithelial lineage. After three washes in phosphate-buffered saline, species-appropriate Alexa Fluor-conjugated secondary antibodies were added for 1 hour at room temperature, and DAPI was used to stain nuclei. Digital micrographs were obtained on an Olympus fluorescence microscope (Tokyo, Japan). Fluorescence signal intensity was subjected to semi-quantitative analysis in ImageJ (NIH, Bethesda, MD, United States) and results were reported as fold change normalized to the control group.
Total cellular RNA was isolated using TRIzol according to the manufacturer’s instructions, and concentration together with the A260/A280 purity ratio were monitored on a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific). Qualified RNA (1 μg) was used as template for first-strand cDNA synthesis with the PrimeScript RT reagent kit with genomic DNA elimination steps. Quantitative amplification was performed on a Roche LightCycler 480 platform using TB Green Premix Ex Taq chemistry. Cycling parameters included a 30-second hot start at 95 °C, followed by 40 cycles of amplification (5 seconds at 95 °C and 30 seconds at 60 °C) with transcript abundance for IL-1β, IL-18, TNF-α, IL-6, and COX-2 being calculated utilizing the comparative 2-ΔΔCt algorithm using glyceraldehyde-3-phosphate dehydrogenase as an endogenous normalizer. Table 1 shows the oligonucleotide primer sequences.
| Gene | Forward primer (5′-3′) | Reverse primer (5′-3′) | Product size (bp) |
| IL-1β | 5′-ATGATGGCTTATTACAGTGGCAA-3′ | 5′-GTCGGAGATTCGTAGCTGGA-3′ | 132 |
| IL-18 | 5′-TCTTCATTGACCAAGGAAATCGG-3′ | 5′-TCCGGGGTGCATTATCTCTAC-3′ | 145 |
| TNF-α | 5′-CCTCTCTCTAATCAGCCCTCTG-3′ | 5′-GAGGACCTGGGAGTAGATGAG-3′ | 220 |
| IL-6 | 5′-ACTCACCTCTTCAGAACGAATTG-3′ | 5′-CCATCTTTGGAAGGTTCAGGTTG-3′ | 149 |
| COX-2 | 5′-CTGGCGCTCAGCCATACAG-3′ | 5′-CGCACTTATACTGGTCAAATCCC-3′ | 183 |
| MMP-9 | 5′-TGTACCGCTATGGTTACACTCG-3′ | 5′-GGCAGGGACAGTTGCTTCT-3′ | 97 |
| GAPDH | 5′-GGAGCGAGATCCCTCCAAAAT-3′ | 5′-GGCTGTTGTCATACTTCTCATGG-3′ | 197 |
Each experimental condition was assayed in triplicate, and each experiment was independently repeated a minimum of three times. Quantitative data are shown as mean ± SEM. Differences between groups were assessed using one-way analysis of variance adjusted for multiple comparisons with Tukey’s honestly significant difference. Statistical sig
The expression of NLRP3 was stable and low in both control and OC groups. DCA stimulation remarkably enhanced the NLRP3 fluorescence intensity, reaching to about 4.8-fold baseline (P < 0.01), which were partly or strongly eliminated by ASP-L and ASP-H treatment, decreased to around 3.3-fold and 1.7-fold compared with DCA implication respectively (P < 0.05). Nigericin (3.8-fold) partially reversed the inhibitory action of ASP-H. All conditions were confirmed as colonic epithelial identity through CK20 co-expression (Figure 1).
DCA treatment resulted in an increase of ASC fluorescence intensity to approximately 5.1-fold with perinuclear speck pattern. ASP-L and ASP-H decreased ASC expression by about 3.5-fold and 2.0-fold, respectively (P < 0.05 vs DCA). Nigericin partially restored ASC speck formation (3.4-fold), suggesting that ASP downregulates ASC oligomerization was due to a dose-dependent manner (Figure 2).
DCA caused a strong induction of cleaved caspase-1 to about 5.6-fold (P < 0.05). ASP-L and ASP-H decreased this to approximately 3.9-fold and 1.8-fold respectively. Nigericin markedly restored caspase-1 activation, confirming that ASP inhibits the entire NLRP3/ASC/caspase-1 inflammasome pathway in a dose-dependent manner (Figure 3).
MCC950 co-treatment significantly reduced cleaved caspase-1 to approximately 1.9-fold in ASP-treated cells. Nigericin increased cleaved caspase-1 to approximately 3.4-fold in ASP-treated cells (P < 0.05), which lent direct pharmacological evidence that the specific mechanism of action of anti-inflammatory effects caused by ASP was NLRP3 inflammasome (Figure 4).
NLRP3 activation by ATP (5 mmol/L) through P2X7 receptor-mediated potassium efflux caused a massive increase of IL-1β, to about 3.6-fold of control (P < 0.01). Treatment with the caspase-1 inhibitor VX-765 led to a nearly 1.3-fold reduction in IL-1β (P < 0.05), establishing caspase-1 as an essential actory downstream of this inflammatory signaling cascade (Figure 5).
Combined NLRP3 stimulation with nigericin and ATP synergistically augmented cleaved caspase-1 to nearly 3.6-fold (P < 0.01). VX-765 was also found to significantly decrease caspase-1 to basal levels (approximately 1.2-fold; P < 0.05) even under dual-infection stimulation conditions, definitively demonstrating that the inflammatory inhibitory mechanism of ASP converges onto NLRP3-dependent signaling pathways leading to decreased processing of caspase-1 (Figure 6).
DCA markedly upregulated all six inflammatory mediators, while ASP decreased them effectively with reversal by nigericin and re-attenuation (CX-09 and VX-765) of effects (Figure 7). IL-6 and MMP-9 were also susceptible to inhibition by CY-09, but their reduced expression was less responsive to VX-765, suggesting that there are partial caspase-1-independent NLRP3 effector mechanisms for these two targets, while IL-1β, TNF-α, and COX-2 exhibit predominant regulation in a caspase-1-dependent manner.
Collectively, these cumulative findings provide evidence that ASP inhibits NLRP3 assembly at or upstream of this event to downregulate four (i.e. IL-1β, IL-18, TNF-α) or all six (TNF-α + IL-6) downstream inflammatory mediators regulated by ASC specks and caspase-1 activation. Nevertheless, the exact molecular target of ASP, that is whether transcriptional inhibition of NLRP3, signaling constituent involved in nuclear factor kappa-B (NF-κB) priming or interfering with canonical triggering of activation such as potassium efflux or mitochondrial ROS, cannot be deduced from the current data set and will require investigation in subsequent studies that use models capable to harvest untangling information like NLRP3-KO mice assessment; ROS scavenging assays and potassium flux assessment (Figure 8).
For the first time, we reveal strong evidence that ASP, a major bioactive macromolecular component derived from Angelica sinensis, protects against DCA-induced inflammatory damage and early neoplastic transformation in human colonic epithelial cells via specific suppression of the NLRP3/caspase-1/IL-1β signaling axis. This well-coordinated approach employing immunofluorescence, reverse-transcription PCR (RT-qPCR), ELISA, and pharmacological pathways’ modulations highlights the molecular mechanism underlying ASP’s chemopreventive properties could spur advancing our understanding of its potential therapeutic capacity in constipative colorectal carcinogenesis.
In this transition from constipation to CRC, the NLRP3 inflammasome has been increasingly recognized as a principal contributor of chronic inflammation. Figures 1-3 show that DCA strongly activates the entire NLRP3/ASC/caspase-1 cascade (immunofluorescence), with 4.8-fold to 5.6-fold increases in intensity (above basal) in NCM460 cells with DCA treatment, whereby ASC is a marker of both NLRP3 activation and caspase-1 assembbly within this protein complex in resting epithelial cells. This result is in line with earlier reports demonstrating that secondary bile acids induce NLRP3 inflammasome signaling in colonic epithelial cells via mtROS generation, potassium efflux and lysosome destabilization-dependent cathepsin B release[6,21,22]. Notably, we demonstrate that ASP not only reduced levels of NLRP3 and ASC but also suppressed cleaved caspase-1 expression in a dose-dependent manner with high doses returning levels to near baseline. The corresponding inhibition of the three inflammasome components indicates that ASP inhibits the pathway upstream (NLRP3 sensor), leading to downstream effects on ASC speck formation and caspase-1 autocleavage.
One of the main conclusions derived from this study is a strong pharmacological validation of the signaling pathway (Figures 4-6). In addition, the finding that nigericin, a prototypical NLRP3 agonist that promotes potassium efflux, counteracted ASP-mediated inhibition of caspase-1 and inflammatory mediators represents compelling evidence that ASP targets NLRP3-mediated signaling directly. The NLRP3-specific inhibitor MCC950[23] further inhibited the caspase-1 activation induced by nigericin in ASP-treated cells (Figure 4), supporting that this is a specific pathway. The caspase-1 specific inhibitor VX-765 amply inhibited activation of caspases-1 and maturation of IL-1β, whereas the addition of ATP (which stimulates NLRP3 by means of P2X7 receptor-dependent potassium efflux) enriched inflammatory signaling (Figure 5). These interdependent effects constitute compelling evidence that the anti-inflammatory mechanism of ASP is driven by NLRP3-caspase-1 axis rather than NLRP3-independent inflammasome pathways (e.g., NLRC4 or AIM2)[24,25]. The redundant modulation experiment (Figure 6) additionally confirmed that even the dual NLRP3 stimulus paradigm (nigericin + ATP) was completely abrogated by caspase-1 blockade, establishing caspase-1 as the crucial degradation bottleneck of the pathway.
In our RT-qPCR and ELISA analyses (Figures 7 and 8), we demonstrate that ASP inhibits a wide array of inflammatory mediators produced downstream or as secondary targets by NLRP3 inflammasome activation including IL-1β, IL-18, TNF-α, IL-6, COX-2, and MMP-9 which were previously shown to play roles in contributing to the constipation-to-CRC continuum. Caspase-1 directly cleaves pro-phagocyte IL-1β and IL-18 from the innate immune response, which serves as important regulators of that process in the intestinal mucosa[13]. TNF-α is induced subsequently by IL-1β driven NF-κB activation and enhances the local inflammatory environment through positive feedback loops in colonic epithelial cells that promote genomic instability[14]. IL-6 initiates STAT3 signaling, a key regulator of CRC cell survival and proliferation[26,27]. COX-2 is the key enzyme in prostaglandin E2 biosynthesis that promotes tumor angiogenesis, immune escape and anti-apoptosis; importantly based on limited evidence, chemopreventive effect of aspirin and other nonsteroidal anti-inflammatory drugs for CRC particularly due to COX-2 inhibition[28,29]. MMP-9 breaks down the extracellular matrix and basement membrane, therefore promoting tumor invasion and metastasis[30]. Through simultaneous inhibition of all six mediators, ASP may globally dampen the chronic inflammatory environment that promotes the further progression from constipation to CRC.
Our data show, interestingly, that different mediators contribute through caspase-1-dependent or -independent mechanisms. Although IL-1β, IL-18, TNF-α and COX-2 were strongly reduced by both CY-09 and VX-765 treatment, greater inhibition of IL 6 and MMP-9 secretion was observed with CY-09 compared to VX-765. This implies that regulation of both IL-6 and MMP-9 may in part depend on caspase-1-independent effector mechanisms of NLRP3, such as pore formation by gasdermin D or the resultant ionic flux[12,31], or through NLRP3-associated kinase signaling that is separate from canonical inflammasome function. In contrast, COX-2 was more potently induced by VX-765 than CY-09 at both mRNA and protein levels, suggesting that its transcriptional upregulation is predominantly dependent on caspase-1–dependent release of IL-1β consistent with the known role of IL-1β/IL-1R1 signaling in mediating COX-2 expression through downstream activation of NF-κB[28,32]. These nuances bear important influence on the concept and design of chemopreventive strategies specific to constipation-associated CRC; rather, targeting the common NLRP3 sensor may yield a broader effect than strategies directed toward caspase-1 effector alone.
The agreement between mRNA and protein data for the majority of mediators reported further supports the validity of our results. However, some quantitative differences for RT-qPCR and ELISA results were observed, notably for IL-18, where proportionally DCA induced a greater protein than mRNA response would indicate. This may be indicative of post-transcriptional regulatory mechanisms, as con-stitutive pro-IL-18 expression by colonic epithelial cells is well established, and caspase-1activa-tion is a rate-limiting step for mature IL-18 release that can proceed without the synthesis of new mRNA[9,25]. These differences are to be expected and highlight the need to integrate transcriptional and translational data, especially for inflammasome-processed cytokines.
It is informative to compare the inhibitory activity of ASP to other naturally occurring compounds with reported activity as suppressors of NLRP3 in either colorectal or inflammatory models. Among this, curcumin primarily inhibits NLRP3 activation via NF-κB inhibition and direct ROS scavenging, while resveratrol inactivates NLRP3 ATPase activity as well as ASC oligomerization. In contrast to those lipophilic small molecules, the water-soluble macromolecular polysaccharide ASP has a unique pharmacological profile; its safety is favorable, it can be orally administered in classical Danggui formulae, and it simultaneously inhibits NLRP3 expression, ASC assembly and caspase-1 cleavage indicating involvement at multiple structural levels of inflammasome action. These combinations require further standardized in vivo validation via direct head-to-head comparisons, but the mechanistic breadth demonstrated here indicates that ASP is a strong contender for competing with existing natural product NLRP3 inhibitors for utility in CRC associated with constipation.
There are several limitations to the current study that should be recognized. First, our experiments were performed using an in vitro cell culture model exclusively with NCM460 cells, limiting the recapitulation of the complex multi
In summary, the results of this study show for the first time that ASP is a potent inhibitor of NLRP3/caspase-1/IL-1β signaling in colonic epithelial cells that attenuates several pro-inflammatory and pro-tumorigenic mediators under DCA-induced stress. These mechanistic findings raise the possibility of ASP being a good candidate chemopreventive agent for CRC associated with constipation. The next priority steps include (1) Validation in a murine model of constipation associated CRC such as the azoxymethane/dextran sodium sulfate protocol plus loperamide-induced constipation; and (2) Pharmacokinetic profiling after oral administration of ASP to determine intestinal bioavailability and colonic tissue exposure for clinical translation.
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