Molecular Mechanisms and Signaling Pathways of the Anti-inflammatory Effects of Functional Foods

Inhibitory Effects of Differentially Cultivated Carrots on AOM/DSS-Induced Colorectal Cancer Via Modulation of Inflammation, Apoptosis, and Gut Microbiota

2026-02-02T09:10:16+00:00

Background: Carrot (Daucus carota) is a widely consumed vegetable rich in dietary fibre, vitamins, minerals, and antioxidant compounds. Carrots contain fibre and bioactive phytochemicals that have been associated with anti-inflammatory and anti-proliferative activities in preclinical settings. Beta-carotene, a prominent carotenoid in carrots, has been reported to modulate immune-related pathways in experimental models; however, the extent to which these observations translate to reduced colorectal cancer risk in humans remains to be determined.

Objective: This study investigated whether cultivation methods influence the bioactivity of carrot extracts in an azoxymethane/dextran sodium sulfate (AOM/DSS)-induced murine model of colitis-associated colon tumorigenesis, with emphasis on apoptosis-related markers, inflammatory responses, and gut microbiota alterations.

Methods: Six-week-old male C57BL/6 mice (~20 g) were maintained under SPF conditions (12-h light/dark cycle; 23 ± 2 °C; 50 ± 5% humidity). After one week of acclimatisation with ad libitum food and water, mice were randomly allocated to six groups (n = 10/group). The normal and AOM/DSS control groups received saline, whereas the Conventional Fertiliser Carrot (CFC), Seawater Fertiliser Carrot (SFC), Trace Element Fertiliser Carrot (TFC), and Deep-sea Water Mineral fertiliser carrot (NFC) groups received the respective carrot extracts (50 mg/mL). Total phenolic and total flavonoid contents were quantified and analysed using GraphPad Prism 9.4.1.

Results: Among the cultivation conditions tested, carrots produced with deep-sea water mineral fertiliser (NFC) were associated with increased expression of apoptosis-related genes and proteins in colon tissue, including p53, p21, Bim, Bad, Bax, Bak, Caspase-9, and Caspase-3. NFC treatment was also associated with lower levels of pro-inflammatory cytokines and mediators (TNF-α, IL-1β, IL-6, IFN-γ, NF-κB, and iNOS) measured across serum, spleen cells, and liver tissues. Gut microbiota profiling indicated a compositional shift in carrot-treated mice relative to controls, including increased relative abundance of taxa previously linked to anti-inflammatory or anti-tumour-associated profiles (e.g., Lachnospiraceae and Mucispirillum schaedleri).

Conclusion: In this AOM/DSS mouse model, carrot extracts—particularly those derived from carrots cultivated with deep-sea water mineral fertiliser—were associated with reduced tumour-related endpoints alongside coordinated changes in apoptotic and inflammatory markers and gut microbiota composition. These findings support further investigation into cultivation-dependent differences in carrot bioactivity and encourage additional studies to clarify mechanisms and assess translational relevance in humans.

Bioactivity of Mineral-Enriched Organic Carrots: Anticancer Effects in HT-29 Cells and Anti-Inflammatory Activity in Mice Splenocytes

2026-02-02T09:16:29+00:00

Carrot (Daucus carota) is a globally important root vegetable with diverse biological activities. Currently, organic cultivation places a greater focus on ecosystem health by using natural organic fertilisers and organic pesticides, such as animal and plant-based fertilisers and natural insecticides, reducing reliance on chemical substances, promoting soil health and ecological balance, and providing healthier and environmentally friendly agricultural products. However, research on the anti-inflammatory properties of carrots, especially comparing traditionally cultivated and organically grown varieties, remains limited. Therefore, this study examined the anticancer and anti-inflammatory properties of carrots cultivated using Natural Dream Cultivation (NC), an organic farming method enhanced with mineral supplementation. The inhibitory effect of NC carrot extracts on HT-29 human colon cancer cells was assessed via MTT assay, while qPCR was used to evaluate mRNA expression of cell cycle- and apoptosis-related genes. Nitric oxide (NO) concentration was measured using the Griess method. In LPS-induced mouse splenocytes, levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assay, and natural killer (NK) cell activity was evaluated via LDH release assay. Results showed that NC significantly suppressed cancer cell proliferation. It up-regulated the expression of cell cycle arrest genes (p53, p21) and pro-apoptotic genes (Bim, Bad, Bax, Bak, caspase-9, caspase-3), while down-regulating anti-apoptotic genes (Bcl-2, Bcl-xL). Furthermore, NC treatment inhibited NO production and reduced the release of inflammatory cytokines (TNF-α, IL-6, IL-1β, IFN-γ, IL-12) in LPS-stimulated splenocytes. NC also promoted NK cell activation. By elucidating the inhibitory effects on cancer cells and modulation of inflammatory responses, this study reveals potential mechanisms behind the anticancer and anti-inflammatory activities of carrots. The innovative mineral-enhanced organic cultivation method presented here offers new possibilities for developing carrots as a functional food candidate supported by in vitro and ex vivo bioactivity relevant to cancer- and inflammation-associated pathways. These findings may inform the future design of dietary research hypotheses and preclinical studies, pending validation in animal models and clinical settings.

Protective Effects of Shoumei (Camellia sinensis) Polyphenols against Hepatic Injury via Antioxidant Mechanisms

2026-02-02T09:20:42+00:00

Tea is globally the second most consumed beverage after water, and its pharmacological properties are extensively documented. It has potent neuroprotective, free radical scavenging, antioxidative, antioncogenic, hepatoprotective, anti-diabetic, antiviral, and chemopreventive properties. This study investigated the hepatoprotective effects of polyphenols extracted from Shoumei, a slightly fermented white tea (Camellia sinensis). HPLC analysis identified gallic acid, catechin, hyperoside, and sulfuretin as the main phenolic constituents. In an in vitro model, Shoumei polyphenols (SPs) attenuated H₂O₂-induced oxidative damage in human normal hepatic L-02 cells. In an in vivo model of CCl₄-induced liver injury in mice, SPs effectively reduced the liver index and ameliorated histopathological damage. Biochemical analyses demonstrated that SPs significantly decreased serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), triglyceride (TG), total cholesterol (TC), blood urea nitrogen (BUN), nitric oxide (NO), malondialdehyde (MDA), interleukin-6 (IL-6), interleukin-12 (IL-12), tumour necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ). Conversely, SPs increased serum levels of albumin (ALB), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). At the molecular level, qRT-PCR and Western blot analyses revealed that SPs up-regulated the mRNA and protein expression of neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase (eNOS), manganese-SOD (Mn-SOD), copper/zinc-SOD (Cu/Zn-SOD), CAT, and inhibitor of nuclear factor kappa B alpha (IκB-α), while down-regulating the expression of inducible nitric oxide synthase (iNOS) and NF-κB p65. The preventive effect of SPs against CCl₄-induced liver injury was comparable to that of silymarin. These results indicate that the identified polyphenols, primarily through their antioxidant capacity, mediate significant hepatoprotection, suggesting Shoumei polyphenols are high-quality natural agents for liver protection.

Antioxidant and Immunomodulatory Effects of Lactobacillus plantarum ZS62 in a Mouse Model of Colitis

2026-02-02T09:33:39+00:00

Lactobacillus plantarum belongs to the genus Lactobacillus, which is the largest genus of lactic acid bacteria. Previous research revealed that L. plantarum ZS62 has a protective effect on the stomach, and the stomach and intestine are often considered to be two inseparable organs that ensure the normal function of the gut. In this study, a dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mouse model was established to investigate the preventive effect of Lactobacillus plantarum ZS62 against IBD, with a focus on its modulatory roles in oxidative stress and immune responses. The alleviative effect of this strain on IBD was evaluated by assessing colon length, histopathological alterations, serum antioxidant capacity, inflammatory cytokine levels, as well as mRNA and protein expression of relevant genes in colon tissues. The results demonstrated that L. plantarum ZS62 significantly alleviated colon shortening and reduced histological damage in IBD mice. It downregulated serum levels of malondialdehyde (MDA), myeloperoxidase (MPO), interleukin-1β (IL-1β), IL-6, IL-12, tumour necrosis factor-α (TNF-α), and interferon-γ (IFN-γ), and decreased the relative mRNA and protein expression of IL-1β, IL-12, TNF-α, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and nuclear factor-κB p65 (NF-κB p65) in colon tissues. Conversely, L. plantarum ZS62 upregulated serum levels of catalase (CAT), total superoxide dismutase (T-SOD), and IL-10, and enhanced the expression of copper/zinc superoxide dismutase (Cu/Zn SOD), manganese superoxide dismutase (Mn SOD), glutathione peroxidase (GSH-Px), CAT, IL-10, and inhibitor of κB-α (IκB-α) at both transcriptional and protein levels in the colon. In summary, Lactobacillus plantarum ZS62 exerted a notable preventive effect against DSS-induced IBD by modulating oxidative stress and inflammatory pathways, highlighting its potential as a probiotic candidate for intestinal health management.

Protective Mechanism of Lactobacillus plantarum KFY02 Preventing CCl₄-Induced Liver Injury by Transforming Geniposide into Genipin to Boost Antioxidant Defence

2026-02-02T09:37:52+00:00

Lactobacillus plantarum belongs to the genus Lactobacillus and is a lactic acid bacterium commonly found in the fermented products of milk, meat, and many vegetables. It can pass through the stomach and colonise the intestine to play a beneficial role. L. plantarum has a wide range of applications in various diseases, but there are few reports on its role in liver injury and as an antioxidant. This study investigated the mechanism by which Lactobacillus plantarum KFY02 (KFY02) alleviates CCl₄-induced liver injury in mice, focusing on its role in enhancing systemic antioxidant capacity through the bioconversion of geniposide to genipin. Experimental results confirmed that KFY02 partially converts geniposide into genipin in vitro. In the mouse model of liver injury, both KFY02 and geniposide significantly reduced the liver index and decreased serum levels of AST, ALT, TG, MDA, and the pro-inflammatory cytokines IL-6, IL-12, TNF-α, and IFN-γ, while up-regulating the activity of antioxidant enzymes SOD and GSH-Px. At the molecular level, KFY02 and geniposide up-regulated the mRNA and protein expression of Mn-SOD, Cu/Zn-SOD, CAT, GSH-Px, and IκB-α, and down-regulated the expression of COX-2, iNOS, and NF-κB in liver tissues. In summary, L. plantarum KFY02 mitigates CCl₄-induced hepatic damage by potentiating the body's antioxidant defences, an effect mediated through the microbial conversion of geniposide to its more active form, genipin.

Process Optimisation of Antioxidant Shuidouchi and Its Protective Effects against DSS-Induced Colitis in Mice

2026-02-02T09:42:35+00:00

Intestinal health plays an important role in maintaining a high quality of life. Digestive absorption, metabolism, and mucosal immunity can decline with age, and the number of probiotics in the digestive system can rapidly decrease. Shuidouchi, a traditional Chinese fermented soybean product, involves multistep production processes that have not yet been standardised. By optimising fermentation parameters and enhancing the content of bioactive components, it can be developed into high-quality products with improved health-beneficial properties. In this study, single-factor experiments were conducted to establish different processing conditions, aiming to identify those yielding the highest levels of active ingredients and the strongest in vitro antioxidant activity. Additionally, the preventive effect of Shuidouchi against dextran sulfate sodium (DSS)‑induced colitis in mice was investigated. The results demonstrated that the optimal process conditions consisted of 12 h of soaking, fermentation in a glass container at 35 °C for 48 h. Shuidouchi produced under these conditions exhibited the highest concentration of soybean isoflavones and displayed superior antioxidant capacity compared to other fermentation conditions. The extract prepared from Shuidouchi obtained via the optimised process (soaked in twice the volume of water for 12 h, placed in a glass container, and fermented at 35 °C for 48 h) prevented colitis-associated shortening of the colon and increased the colon weight‑to‑length ratio in mice.

Furthermore, Shuidouchi extract significantly reduced the disease activity index and decreased serum levels of endothelin (ET), substance P (SP), and interleukin‑10 (IL‑10), while elevating levels of somatostatin (SS), vasoactive intestinal peptide (VIP), and interleukin‑2 (IL‑2) in colitic mice. It also increased colon tissue levels of glutathione (GSH) and superoxide dismutase (SOD), whereas it reduced myeloperoxidase (MPO) and malondialdehyde (MDA) levels. Gene expression analysis in colon tissues revealed that Shuidouchi extract upregulated the mRNA expression of neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase (eNOS), c‑Kit, and stem cell factor (SCF). Moreover, Shuidouchi treatment downregulated the protein expression of interleukin‑6 (IL‑6), IL-12, and tumour necrosis factor‑α (TNF‑α) in mice with colitis. In conclusion, the optimised Shuidouchi processing method effectively alleviates experimental colitis, an effect closely associated with its elevated soybean isoflavone content. The results of this study can confirm the effect of Shuidouchi on colitis only to a certain extent. Therefore, it is necessary to further verify the effect of Shuidouchi on the human body. At the same time, the mechanism of Shuidouchi in the human body also requires comprehensive and in-depth experiments. As basic research, this study has played a specific theoretical role.

Hepatoprotective Effects of Liubao Tea Polyphenols against CCl₄-Induced Liver Injury in Mice via Antioxidant Activity

2026-02-02T09:47:11+00:00

Active oxygen free radicals cause oxidative stress, which is a common pathophysiological mechanism of liver diseases. Tea polyphenols can sequester lipid peroxidation free radicals during the peroxidation process, lower polyphenolic free-radical content, and interrupt free-radical oxidation chain reactions, thereby effectively removing free radicals. This study investigated the protective effect of Liubao tea polyphenols (PLT) against carbon tetrachloride (CCl₄)-induced liver injury in mice. Mice were pretreated with PLT prior to the administration of CCl₄ (10 mL/kg) to induce hepatic damage. Subsequently, liver and serum biochemical parameters, along with the expression levels of relevant messenger RNAs (mRNAs) and proteins in liver tissue, were assessed. The results demonstrated that PLT administration significantly ameliorated liver injury, as indicated by reduced liver indices and improved histological architecture. Specifically, PLT downregulated serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TGs), and malondialdehyde (MDA), while elevating the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Furthermore, PLT decreased the serum concentrations of pro-inflammatory cytokines, including interleukin-6 (IL-6), IL-12, tumour necrosis factor-α (TNF-α), and interferon-γ (IFN-γ). Histopathological examination revealed that PLT attenuated CCl₄-induced central venous alterations and hepatocellular damage. At the molecular level, quantitative PCR and Western blot analyses confirmed that PLT upregulated the mRNA and protein expression of antioxidant enzymes—Cu/Zn-SOD, Mn-SOD, catalase (CAT), and GSH-Px—as well as the NF-κB inhibitor IκB-α in liver tissues. Conversely, PLT downregulated the expression of cyclooxygenase-2 (COX-2) and nuclear factor kappa B (NF-κB). Additionally, PLT increased the protein levels of phosphorylated NF-κB p65 (p-NF-κB p65) and cytochrome P450 reductase in injured livers. PLT comprises several bioactive compounds, such as gallic acid, catechin, caffeine, epicatechin (EC), epigallocatechin gallate (EGCG), gallocatechin gallate (GCG), and epicatechin gallate (ECG), which may contribute to its broad biological activities. In conclusion, PLT exerts a preventive effect against CCl₄-induced liver injury, comparable to that of silymarin, likely through the modulation of oxidative stress and inflammatory pathways. In this study, toxic carbon tetrachloride was used to simulate chemical-induced liver injury, and the observed effects remained at the laboratory level. In order to better prove this study’s argument, future research on the human body is expected. The role of PLT in liver injury needs to be further studied, which will be conducive to more obvious discoveries of the link between its active components and their mechanisms. At the same time, in view of the mechanism of PLT, it is necessary to verify the mechanism more accurately for the differences across PLT components in the future.

Protective Effects of Lactobacillus fermentum CQPC08 against 4-Nitroquinoline-1-Oxide–Induced Tongue Cancer in Mice

2026-02-02T09:58:01+00:00

Tongue cancer is a malignant tumour originating in the anterior part of the tongue and is one of the most common malignant tumours in the oral and maxillofacial region. Lactic acid bacteria (LAB) have been of great importance in preserving vegetables, since the fermentation process results in lactic acid production, leading to a decrease in pH, which makes the environment unsuitable for the growth of microorganisms and pathogens that cause spoilage. This study investigated the in vivo chemopreventive efficacy and mechanistic actions of Lactobacillus fermentum CQPC08 (LF-CQPC08), a novel strain isolated from traditional Sichuan pickled vegetables, against experimental tongue cancer. A mouse model of tongue carcinogenesis was established using the carcinogen 4-nitroquinoline 1-oxide (4-NQO), with the commercial strain Lactobacillus delbrueckii subsp. bulgaricus serving as a comparative positive control. Biochemical analyses of serum and tissue samples revealed that LF-CQPC08 administration significantly mitigated the tumour-associated decline in systemic immunity. This was evidenced by the attenuation of reduced splenic and thymic indices, enhanced macrophage phagocytic function, and elevated serum levels of immunomodulatory factors, including granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), immunoglobulins (IgG, IgM), interleukin (IL)-4, IL-12, tumour necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ). Furthermore, LF-CQPC08 treatment ameliorated oxidative stress in tongue tissues, as indicated by increased activities of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), coupled with a decrease in malondialdehyde (MDA) levels. At the molecular level, quantitative PCR analysis demonstrated that LF-CQPC08 upregulated the mRNA expression of cytoprotective and pro-apoptotic genes—nuclear factor erythroid 2–related factor 2 (Nrf2), heme oxygenase-1 (HO-1), glutathione S-transferase pi (GST-π), and Bcl-2-associated X protein (Bax). Conversely, it downregulated the expression of tumour-associated genes p53, p63, p73, phosphatase and tensin homolog (PTEN), and the anti-apoptotic genes B-cell lymphoma 2 (Bcl-2) and B-cell lymphoma-extra large (Bcl-xL). Collectively, these findings suggest that LF-CQPC08 attenuates 4-NQO–associated pathological and molecular changes in a mouse model, potentially via immunomodulatory, redox, and apoptosis-related pathways. Further studies (including validation of tumour endpoints and human studies) are required to assess translational relevance.

Hepatoprotective Effects of Lotus Leaf Flavonoids in CCl₄-Induced Liver Injury in Mice

2026-02-02T10:03:24+00:00

Modern medical studies have revealed that the main active components of lotus leaves are flavonoids and alkaloids. The flavonoids in lotus leaves have been reported to reduce the blood lipid level, to have anti-allergy, anti-cancer, anti-ageing, bacteriostatic, and anti-oxidation functions, and to be helpful in treating cardiovascular diseases. This study examined the protective effects of lotus leaf flavonoids (LLF) against carbon tetrachloride (CCl₄)-induced liver injury in mice. A Kunming mouse model of hepatic injury was established via intraperitoneal injection of CCl₄. The mice were orally administered LLF, after which serum biochemical markers and hepatic mRNA expression levels were analysed. Compared with the model group, LLF treatment significantly lowered the liver index and reduced serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and total cholesterol (TC). Histopathological evaluation further demonstrated that LLF effectively attenuated morphological disruption and hepatocellular necrosis in liver tissues exposed to CCl₄. Quantitative polymerase chain reaction (qPCR) analysis revealed that LLF significantly up-regulated the mRNA expression of antioxidant enzymes, including copper/zinc superoxide dismutase (Cu/Zn-SOD), manganese superoxide dismutase (Mn-SOD), and catalase (CAT), while down-regulating the expression of pro-inflammatory mediators such as tumour necrosis factor-α (TNF-α), nuclear factor kappa B (NF-κB), and interleukin-1β (IL-1β) (p < 0.05). In conclusion, LLF exhibits notable hepatoprotective activity against CCl₄-induced liver injury and holds promise for further development as a natural therapeutic agent. Lotus and its leaves have certain biological activities, in particular, some studies showed that they had good antioxidant activity, and their antioxidant activity also played a protective role in cells, especially in liver cells and the liver. This study also showed that the polyphenols contained in lotus leaves had a good intervention in alcoholic liver injury.