A fluorescent aptamer sensor for kanamycin was constructed based on G-quadruplex DNAzyme and Exo III-assisted signal amplification. Target binding triggered cDNA release, activating Exo III to cleave HP and release G-quadruplex, which formed a DNAzyme with hemin to catalyze OPD oxidation and generate fluorescence. The method achieved a detection limit as low as 0.063 nM.

Objective To compare 26-h glycemic responses to uninterrupted sitting versus hourly brisk walking breaks and an alternating walking–resistance break strategy in sedentary young adults, with exploratory sensitivity analyses adjusting for energy expenditure (EE). Methods Eighteen sedentary, healthy young adults (11 female; age, mean (SD)=23.7 (2.5) years; BMI, 21.2 (2.0) kg/m2) completed three 26-h laboratory protocols in a randomized crossover design. Each protocol included a 22-h stay in a metabolic chamber and one 9-h intervention: uninterrupted sitting (SIT), 8-min bouts of brisk walking at 60% VO2max every 60 min (WALK), or alternating 8-min bouts of simple resistance activities and brisk walking every 60 min (RESWALK). Interstitial glucose was recorded every 5 min by continuous glucose monitoring across the all three 26-h trials; the primary outcome was 26-h glucose incremental area under the curve (iAUC). Results The primary outcome, 26-h glucose iAUC, showed a significant condition effect (overall p=0.039). Compared with SIT, RESWALK reduced 26-h iAUC by 17.3% (Cohen’s d=−0.61, 95% CI −1.09 to −0.12; p=0.043), while the reduction in WALK was not significant; these effects were attenuated after adjusting for EE (p=0.085). For secondary outcomes, both WALK and RESWALK reduced glucose iAUC during the 9-h intervention window (p≤0.027), with RESWALK being more effective than WALK (p=0.021). However, during the 5-h evening period, iAUC was higher in WALK compared with SIT (p=0.011), while no differences were observed during the 8-h sleep period. Regarding glucose variability, only CONGA_1 was significantly lower in RESWALK versus SIT (p≤0.041). Conclusions Compared with prolonged sitting, interrupting sedentary behavior with hourly breaks of combined walking and resistance exercises improves 26-h glycemic control in healthy young adults, EE-adjusted sensitivity analyses attenuated the 26-h effect. Walking-only breaks provided daytime benefits but did not significantly improve 26-h glucose exposure. Trial registration ChiCTR1800019120.

Gallic acid (GA), also known as 3,4,5-trihydroxybenzoic acid, is a polyphenolic bioactive component in pomegranates, grapes, tea leaves, and gallnuts. This study employed LC-MS/MS to systematically analyze the pharmacokinetic and tissue distribution of GA in rats. Our findings demonstrated that GA accumulated most prominently in kidney and prostate tissues. Based on these targeted distribution characteristics, chronic bacterial prostatitis (CBP) was chosen as a target tissue inflammatory model to explore the anti-inflammatory effects of GA. The results showed that GA exerted a direct anti-inflammatory effect by inhibiting the activation of the NF-κB inflammatory signaling pathway and reducing the release of pro-inflammatory cytokines such as IL-6, IL-1β, and TNF-α in both lipopolysaccharide (LPS)-induced RWPE-1 cell model and CBP rats. Untargeted prostate tissue metabolomics identified 43 metabolites in the prostate as metabolic markers associated with CBP rats. Additionally, mass spectrometry imaging (MSI) was used to visualize the heterogeneous distribution of these metabolites in prostate tissue and reveal the anti-inflammatory metabolic mechanism of GA, which was related to arginine metabolism, linoleic acid metabolism, and the downregulation of arachidonic acid metabolism. In conclusion, GA improves the physiological disorders associated with CBP by exerting anti-inflammatory effects and regulating metabolic homeostasis. These findings may shed new light on our understanding of the application value of natural polyphenols in food function and nutrition.

The integration of nanotechnology into oncology has profoundly reshaped cancer treatment, enabling drug delivery systems with remarkable precision, enhancing antitumor efficacy, and simultaneously addressing major challenges such as cardiotoxicity, one of the most prevalent and serious adverse effects of conventional chemotherapy. This review systematically examines the dual role of nanotechnology, highlighting its capacity to enhance the therapeutic effectiveness of anticancer treatments while concurrently mitigating cardiotoxic side effects. The discussion centers on a broad spectrum of nanocarrier platforms, such as liposome-based, polymeric nanocarriers, and inorganic nanocarriers organized according to their structural features and therapeutic benefits, thereby enabling a systematic comparison with conventional drug delivery strategies. By improving drug bioavailability, enabling controlled release, and achieving precise tumor-specific targeting, these nanocarrier systems enhance antitumor efficacy while concurrently reducing collateral damage to healthy tissues. Moreover, recent preclinical and clinical studies were summarized to demonstrate substantial advances in this interdisciplinary field, while also identifying persistent challenges that remain to be addressed. Finally, the review explores future directions, with particular emphasis on the integration of artificial intelligence to optimize nanocarrier design and the promise of personalized nanomedicine in transforming cancer care. Overall, this work provides a critical foundation for advancing next-generation, patient-tailored cancer therapies.

Introduction: Delayed sleep-wake phase disorder(DSWPD)is a serious threat to the physical and mental health. There are some problems with current clinical treatment methods, and exercise is an alternative to chronotherapy. Therefore, we aimed to study the effects of two different exercise methods, aerobic and resistance, on sleep, melatonin, inflammatory factors and mood in college students with DSWPD. Methods: Male college students aged 18-28 years with DSWPD and no regular exercise habits were recruited to participate in a randomized crossover trial. Three-day moderate-intensity aerobic and resistance exercises were conducted. Sleep quality, urine melatonin concentration, blood inflammatory factors, and mood changes were evaluated. Results: Resistance exercise (RE) improved five sleep indicators (P < 0.05),including sleep onset time, sleep onset latency, total sleep time, wake after sleep onset and sleep efficiency, whereas aerobic exercise (AE) only improved sleep onset time and sleep efficiency (P < 0.05). In addition, RE and AE increased urine aMT6s, IL-10 and decreased IL-6. But RE was more effective in improving sleep onset time, sleep efficiency and urine aMT6s, IL-6. In terms of mood indicators, aerobic and resistance reduced self-rating anxiety scale, but AE also had an improvement effect on self-rating depression scale. Conclusion: Aerobic and resistance exercises can cause male college students with DSWPD to fall asleep earlier, improve sleep quality, increase melatonin concentration, reduce body inflammation, promote the synthesis of anti-inflammatory cytokines, and improve mood. Of the two exercise intervention methods, the effect of RE was more significant.

Galectin-1 (Gal-1), a member of the galectin family, has emerged as a regulator of tumor progression. Several studies have reported the upregulation of Gal-1 expression in multiple cancer cells and its promotion on tumor proliferation. However, the mechanism by which Gal-1 promotes tumor growth remains to be thoroughly understood. In this study, it was discovered that high expression of Gal-1 in various cancers was inversely correlated with the overall survival of patients. Through constructing Gal-1-overexpressing cell lines, it was uncovered that cell proliferation and colony formation were significantly improved. The results of transcriptomic and proximity-labeling-based proteomic analyses indicated that Gal-1 interacted with poly [ADP-ribose] polymerase 1 (PARP1) and histone H1.2 in lung cancer cells. In the case of etoposide treatment leading to DNA double-strand break, Gal-1 accelerated the degradation of H1.2 by enhancing its interaction with PARP1 and promoting its PARylation. It caused the activation of downstream DNA repair pathways such as the serine-protein kinase ATM and nibrin (NBS1) signaling pathways, thus reducing apoptosis, and the Gal-1 inhibitor thiodigalactoside (TDG) could restore cell sensitivity to etoposide. Upon knockdown of Gal-1, DNA damage led to impaired activation of ATM and NBS1 phosphorylation, thereby increasing the sensitivity of the A549 cell line to etoposide. Finally, using a tumor-bearing mouse model, we observed that, in tumors with high Gal-1 expression, the combination treatment of TDG and etoposide significantly inhibited tumor growth. This study provides new clues for the role of Gal-1 in the development of tumors and renders suggestions for the medication of patients with high Gal-1 expression in the clinic.

Background:The aim of this study was to assess the effects of short-term high-intensity interval training (HIIT) and moderate-intensity continuous aerobic training (MICT) with matched energy expenditure on metabolic flexibility and other metabolic parameters in adult male individuals with obesity. Methods:Twenty male individuals with obesity (age: 21.4 ± 1.5 years; body mass index: 31.0 ± 3.6 kg/m2) were enrolled in this crossover design study. Participants were randomly allocated to an intervention sequence of 1) 3 consecutive days of HIIT (30 min, 6 × 2.5 min bouts at 90 % Peak Oxygen Uptake (VO2peak), alternated with 2.5 min active recovery periods [25 % VO2peak]), or 2) three consecutive days of MICT (60 min at 50 % VO2peak), with a washout period of 1 week. Respiratory quotient (RQ), glucose, C-peptide, insulin, and non-esterified fatty acid (NEFA) levels were measured both during fasting and throughout a 180 min oral glucose tolerance test (OGTT), conducted before and after the intervention. The incremental area under the curve (iAUC) and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) were derived. ΔRQ was defined as the average RQ during the OGTT minus fasting RQ. Results:Short-term exercise intervention significantly reduced HOMA-IR in male individuals with obesity (P < 0.001) and concurrently increased the Matsuda index (P = 0.001). Both exercise interventions led to a similar decrease in fasting RQ (P = 0.001), C-peptide, insulin, and HOMA-IR (P < 0.001). Additionally, they reduced the iAUC for glucose and insulin during the OGTT (P < 0.001). The ΔRQ and the RQ iAUC in the HIIT group were significantly higher than those in the MICT group, and different exercise intensities exhibited interactive effects within groups for RQ iAUC and ΔRQ (P = 0.013 and P = 0.012). Conclusions:Two short-term exercise interventions similarly improved glucose tolerance, with HIIT demonstrating a more advantageous effect on metabolic flexibility than MICT in male individuals with obesity. Trial registration:The trial was officially registered at www.chictr.org.cn (ChiCTR2300072884).

BACKGROUND:Tumor-associated macrophages (TAMs) are crucial in hepatocellular carcinoma (HCC) progression and prognosis, making them promising immunotherapy targets. In traditional Chinese medicine (TCM), qi stagnation and blood stasis are linked to the HCC tumor microenvironment (TME), but few studies explore the effects of related TCM herbs on the TME. Calceolarioside B, a key phenylethanoid glycoside in Akebiae Fructus, has not been well studied for its pharmacological activities or molecular targets, and its role in HCC remains unclear. PURPOSE:This study aimed to investigate the effects of Calceolarioside B on TAMs in HCC and clarify its potential targets and regulatory mechanisms. METHODS:Murine intrahepatic transplantation HCC models and macrophage-HCC cell co-culture systems were used to investigate the effects of Calceolarioside B on M2-like TAMs polarization and infiltration, and tumor growth. Cellular thermal shift assay, small molecular pull-down assay and surface plasmon resonance were utilized to identify the potential targets regulating M2-like TAMs. Single-cell RNA sequencing and TCGA dataset analyses clarified the differential expression, prognosis, and TAMs association of the potential targets in HCC. RESULTS:Calceolarioside B reduces M2-like TAMs polarization and infiltration in the TME by binding to and inhibiting matrix metallopeptidase-12 (MMP12) form both macrophages and HCC cells, thereby preventing immunosuppressive effects. Public database analysis revealed that MMP12 overexpression promoted macrophage infiltration, with MMP12+ macrophages preferentially aggregating in primary and metastatic HCC tumors. CONCLUSION:Calceolarioside B is identified as a novel MMP12 inhibitor modulating TAMs in the TME, offering a potential TAM-targeting strategy for HCC therapy.

Vitamin D deficiency is a global public health problem associated with intramyocellular lipid (IMCL) accumulation, leading to insulin resistance. Aerobic exercise improves lipid metabolism, insulin sensitivity, and vitamin D levels. However, the mechanism by which aerobic exercise regulates IMCL remains unclear. C57BL/6J male mice were randomly divided into four groups: control (CON), vitamin D-deficient (VDD), control exercise, and vitamin D-deficient exercise (VDDE). Moreover, we generated skeletal muscle-specific vitamin D receptor (VDR)-knockout (mVDR-/-) mice and classified them into four groups: VDRflox/flox control (FC); mVDR-/-; exercise and VDRflox/flox control; and exercise and mVDR-/- (emVDR-/-). All exercise mice underwent a 12-week aerobic exercise program on a treadmill at speeds progressively increasing from 10 to 17 m/min. The VDD group mice exhibited decreased VDR expression, lipolysis factors (ATGL and Hormone-sensitive lipase (HSL)), and fatty acid oxidation (SIRT1 and PGC1α), and increased expression of lipid synthesis factors (DGAT1 and FATP1) compared with the CON group. Conversely, the VDDE group mice showed a significant increase in VDR, ATGL, HSL, SIRT1 and PGC1α expression, alongside a decrease in DGAT1, DGAT2, FATP1 and CD36 expression compared with the VDD group. Moreover, mVDR-/- mice exhibited impaired lipid metabolism (FATP1, CD36, SREBP1C, DGAT1, DGAT2 and ATGL) and fatty acid oxidation (SIRT1 and PGC1α) compared with the FC group mice. However, emVDR-/- mice did not show improved lipid metabolism or fatty acid oxidation related factors compared with mVDR-/- mice. Therefore, aerobic exercise attenuates IMCL accumulation may by upregulating VDR associated with restored SIRT1/PGC1α signaling mediated lipid metabolism in skeletal muscle.

Herein, a new heterogeneous CoSe2-x@N-C material with abundant selenium vacancies is synthesized via an in-situ carbonization-selenization process from cobaltic metal organic framework (Co-MOF). The obtained CoSe2-x@N-C has a unique electronic structure and rich active sites, which can activate peroxymonosulfate (PMS) to degrade carbamazepine (CBZ) with superior catalytic performance and stability. The quenching experiments and EPR test show that SO4•− is the dominant reactive oxidation species (ROSs) for CBZ degradation. Significantly, systemic electrochemical tests and theoretical calculations illustrated that the dominant role of SO4•− is attributed to the existence of abundant selenium vacancies in CoSe2-x@N-C, which can adjust the density of electron cloud of the Co atoms in CoSe2-x@N-C to improve the PMS adsorption and promoting the conversion of transition metallic redox pairs (Co3+/Co2+). This work provides a facile way to improve the activity and stability of CoSe2 by defect engineering in the PMS based advanced oxidation process (AOPs).

Background Systemic lupus erythematosus is a clinically heterogeneous autoimmune disease that lacks reliable diagnostic biomarkers. In our study, we aimed to identify a novel biomarker for the diagnosis and disease activity monitoring of SLE. Methods Bulk RNA and scRNA-seq datasets were obtained from the Gene Expression Omnibus database. In this study, differential analysis, cell-cell communication algorithm, functional enrichment analysis, human protein map database analysis, protein-protein interaction analysis and immune cell infiltration analysis were utilized to identify the hub genes between SLE and healthy groups. Furthermore, clinical data from 68 SLE patients and 31 healthy controls were collected for verification. Changes in IFITM3 levels were confirmed through quantitative real-time polymerase chain reaction, western blotting, and flow cytometry analyses. Result Bioinformatic analyses showed that IFITM3 expression was significantly upregulated in peripheral monocytes from patients with SLE. IFITM3 mRNA levels showed a significant diagnostic value for SLE, with an AUC value of 87.14%. IFITM3 expression was associated with the systemic lupus erythematosus disease activity index, as well as C3, C4, and IgG levels. The results of Chi-square test showed that those in the IFITM3-positive group had a higher percentage of several clinical manifestations such as thrombocytopenia, leukopenia, low complement, and fever. Conclusions These findings indicated an obviously increased expression of IFITM3 in peripheral blood monocytes of patients with SLE and verified IFITM3 as a promising diagnostic marker for SLE and associated with disease activity.

As a traditional Chinese medicine (TCM), Cortex Periplocae (CP) has a wide range of pharmacological effects, as well as toxic side effects. The main toxic components of it are cardiac glycosides, which tend to cause cardiotoxicity. Currently, it has also been reported in studies to cause hepatotoxicity, but it is not clear whether the hepatotoxicity is related to the toxicity caused by the reactive metabolites. This study aims to investigate the target components of CP that generate reactive metabolic toxicity. The fluorescent probe method was used to detect glutathione (GSH)-trapped reactive metabolites in a co-incubation system of CP extract with rat liver microsomes. Identification of GSH conjugates was performed by LC–MS/MS and that of the possible precursor components that produce reactive metabolites was conducted by UPLC–Q-TOF/MS. Cell viability assays were performed on HepG2 and L02 cells to determine the cytotoxicity of the target components. The findings of our study demonstrate that the extract derived from CP has the ability to generate metabolites that exhaust the intracellular GSH levels, resulting in the formation of GSH conjugates and subsequent cytotoxic effects. Through the utilization of the UPLC–Q-TOF/MS technique, we were able to accurately determine the molecular weight of the precursor compound in CP to be 355.1023. The primary evidence to determining the GSH conjugetes relies on the appearance of characteristic product ions resulting from central neutral loss (CNL) scanning of 129 Da and product scanning of m/z 660 in the positive MS/MS spectrum. Through analysis, it was ultimately ascertained that the presence of chlorogenic acid (CGA) and its isomers, namely neochlorogenic acid (NCGA) and cryptochlorogenic acid (CCGA), could lead to the production of GSH conjugates, resulting in cytotoxicity at elevated levels. Taking these findings into consideration, the underlying cause for the potential hepatotoxicity of CP was initially determined.

Therapeutic agents can be transformed into reactive metabolites under the action of various metabolic enzymes in vivo and then covalently combine with biological macromolecules (such as protein or DNA), resulting in increasing toxicity. The screening of reactive metabolites in drug discovery and development stages and monitoring of biotransformation in post-market drugs has become an important research field. Generally, reactive metabolites are electrophilic and can be captured by small nucleophiles. Glutathione (GSH) is a small peptide composed of three amino acids (i.e., glutamic acid, cysteine, and glycine). It has a thiol group which can react with electrophilic groups of reactive metabolic intermediates (such as benzoquinone, N-acetyl-p-benzoquinoneimine, and Michael acceptor) to form a stable binding conjugate. This paper aims to provide a review on structure-based reactivity profiles of reactive metabolites with GSH. Furthermore, this review also reveals the relationship between drugs' molecular structures and reactive metabolic toxicity from the perspective of metabolism, giving a reference for drug design and development.

The emergence of chemoresistance in cervical cancer is extremely challenging in chemotherapy. Oxidative stress has emerged as the regulatory factor in drug resistance, but the detailed mechanism is still unknown. Stress granules, are membrane-less ribonucleoprotein-based condensates, could enhance chemoresistance by sequestering proapoptotic proteins inhibition of cell death upon exposure to drug-induced oxidative stress. Galectin-7, a member of galectin family, exerts varied roles in tumor repression or progression in different cancers. However, its role in cervical cancer has not been sufficiently studied. Here, we found that galectin-7 promotes cisplatin (CDDP) induced apoptosis and associates with stress granule-nucleating protein G3BP1 degradation. With the treatment of cisplatin, galectin-7 could enhance apoptosis by upregulating cleaved-PARP1 and the generation of reactive oxygen species (ROS), promoting mitochondrial fission, and reducing mitochondrial membrane potential (MMP). Furthermore, galectin-7 also reduces resistance by facilitating cisplatin-induced stress granules clearance through galectin-7/RACK1/G3BP1 axis. All these data suggested that galectin-7 promotes cisplatin sensitivity, and it would be potential target for potentiating efficacy in cervical cancer chemotherapy.

The toxicity and side effects of chemotherapeutic drugs remain a crucial obstacle to the clinical treatment of hepatocellular carcinoma (HCC). Identifying combination therapy from Chinese herbs to enhance the sensitivity of tumors to chemotherapeutic drugs is of particular interest. Astragalus polysaccharide (APS), one of the natural active components in Astragalus membranaceus, has been reported to exhibit anti-tumor properties in diverse cancer cell lines. The aim of this study was to determine the effect of APS on Doxorubicin (Dox)-induced apoptosis in HCC and the underlying mechanism. The results showed that APS dose-dependently promoted Dox-induced apoptosis and enhanced endoplasmic reticulum (ER) stress. Additionally, APS decreased the mRNA level and protein stability of O-GlcNAc transferase (OGT), and increased the O-GlcNAcase (OGA) expression. Furthermore, OGT lentiviral transfection or PugNAc (OGA inhibitor) treatment reversed the ER stress and apoptosis induced by the combination of Dox and APS. A xenograft tumor mouse model confirmed that the combination of APS and Dox showed an advantage in inhibiting tumor growth in vivo. These findings suggested that APS promoted Dox-induced apoptosis in HCC cells through reducing the O-GlcNAcylation, which led to the exacerbation of ER stress and activation of apoptotic pathways.

Terminal N-acetylglucosamine (GlcNAc) N-linked glycosylation is a truncated N-glycosylated modification that has been reported to be involved in various diseases, such as autoimmune diseases, cancers, and neurodegenerative diseases. New and simple tools will be always valuable for further characterization of the functions of this kind of glycosylation. Our previous paper proved that an optimized lectin created from Agrocybe aegerita GlcNAc selective lectin (AANL) named AANL6, can effectively identify O-GlcNAcylation, which is terminal GlcNAc Olinked glycosylation. We speculated that AANL6 could also be used to identify terminal GlcNAc N-linked glycosylation. Using therapeutic monoclonal antibodies as a model of terminal GlcNAc N-glycosylated proteins, we proved that AANL6 could selectively identify terminal GlcNAc N-linked glycosylation. The ratio of terminal GlcNAc N-linked glycosylation was increased by enrichment with AANL6 in human serum. Using cell membrane proteins as a complex sample, we found that AANL6 bound to the sperm surface, which expresses abundant terminal GlcNAc N-glycans, but did not bind to some tumor cell surfaces such A549 and MCF-7 cells, which is rich in high mannose glycoforms. In conclusion, AANL6 was identified as a powerful tool to probe terminal GlcNAc N-linked glycosylation and would be valuable for uncovering the function of this glycosylation.

Ethnopharmacological relevance: Cimicifuga foetida L. is a well-established traditional Chinese medicine with heat-clearing and detoxifying effects and has good therapeutic effect on oral mucosal ulcer and pharyngitis. The rhizome of this herb is rich in triterpenoid glycosides, including 23-O-acetylshengmanol-3-o-alpha-L-arabinoside (DA).Aim of the study: Whether and how DA attenuates acute lung injury (ALI) are unclear. Accordingly, we focused on its anti-inflammatory effects and underlying molecular mechanisms in lipopolysaccharide (LPS)-stimulated ALI mice and RAW264.7 cells.Materials and methods: The model of ALI mice was established by exposed intratracheal instillation of LPS. Lung pathological changes were evaluated by hematoxylin and eosin staining. Pulmonary function was assessed by whole-body plethysmography. Total protein content in bronchoalveolar lavage fluid (BALF) was detected by bicinchoninic acid method. Wet/dry lung ratio was used to evaluate the degree of pulmonary edema in mice. The levels of pro-inflammatory mediators were measured using enzyme-linked immunosorbent assay. The relative expression of pro-inflammatory gene mRNA was examined by RT-qPCR. The expression of inflammatory-related proteins was detected by Western blot. RAW264.7 cells were used to test the anti-inflammatory effects of DA in vitro. Cytotoxicity was assessed using a MTT assay. Nitric oxide production was measured by Griess assay. The production and expression of inflammatory mediators and the protein levels of inflammatory signaling molecules in the NF-Kappa B and MAPK pathways were measured. Furthermore, immunofluorescence staining was used to analyze the expression of p-I Kappa B alpha, p-ERK, and p-p38 in lung macrophages and the nuclear translocation of NF-Kappa B p65 and AP-1 in cells.Results: DA evidently alleviated histopathological changes and ameliorated pulmonary edema. Moreover, DA could reduce excessive inflammatory reaction in lung tissue as manifested by the reduction of proinflammatory mediators (IL-1 beta, IL-6, TNF-alpha, MCP-1, iNOS, and COX-2) in BALF, serum, and lung tissues. Further, DA inhibited the activation of the NLRP3/caspase-1 pathway in the lung. DA reduced the production and expression of the proinflammatory mediators above in RAW264.7 cells. Mechanistically, DA remarkably blocked the nuclear translocation of NF-Kappa B p65, suppressed I Kappa B alpha phosphorylation, and markedly reduced the nuclear translocation of AP-1 and the phosphorylation of ERK and p38.Conclusions: The findings demonstrated that DA exerts anti-inflammatory effects in LPS-stimulated ALI mice and macrophages by downregulating the NLRP3/caspase-1 signaling pathway in lung tissue and the I Kappa B/NF-Kappa B and MAPKs/AP-1 pathways in macrophages, suggesting that DA may be promising in ALI treatment.

BACKGROUND:Berberine has been shown in clinical studies to have many health benefits, including anti-inflammatory and antioxidant properties, along with gut-flora balancing properties. However, its clinical efficacy is hindered by its low oral bioavailability and rapid metabolism.PURPOSE:This study aims to identify the berberine metabolites' forms and characterize their biodistribution patterns in and out of HepG2 cells.METHODS:The qualitative analysis of metabolites of berberine in HepG2 cells was performed using the LC/MSn-IT-TOF method. Subsequent cellular pharmacokinetics characterization of intracellular and extracellular berberine and its metabolites was performed by LC-MS/MS analysis.RESULTS:Berberine's metabolites of phase I metabolism were demethyleneberberine, jatrorrhizine, columbamine, berberrubine, etc., while its phase II metabolites were sulfate and glucuronide conjugates of phase I metabolites. Among the phase I metabolites of berberine, jatrorrhizine+columbamine accounted for over two-thirds of the total, followed by demethyleneberberine, which accounted for about a quarter. The intracellular demethyleneberberine is 25.14 times more enriched than extracellular demethyleneberberine. On the other hand, jatrorrhizine+columbamine and berberrubine were primarily distributed extracellularly, and their extracellular concentrations were 7.13 times and 15.61 times of their intracellular concentrations, respectively. Berberine metabolites produced in phase II metabolism are predominantly sulfate conjugates.CONCLUSION:Our results show that demethyleneberberine is highly concentrated intracellularly in HepG2, possibly because it is an essential metabolite of berberine that likely contributes to berberine's efficacy. In light of our findings, berberine's poor plasma concentration-effectiveness characteristics have been partially explained.