This study investigated the toxicological mechanisms of different processed products of Strychnos nux-vomica seeds using serum metabolomics analysis. Seventy-two female KM mice were randomly divided into six groups (n = 12 per group): control, raw seeds, vinegar-processed seeds, urine-soaked seeds, sand-roasted seeds, and oil-fried seeds. Each processed preparation group received an aqueous decoction of the original herb at the same concentration for 7 days. Hepatic and renal coefficients were measured in each group, and the levels of hepatic and renal injury markers, including aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN), and creatinine (CRE) were determined using a fully automated biochemical analyzer. Hematoxylin-eosin (HE) staining was used for histopathological examination of the liver and kidney. Liquid chromatography-mass spectrometry (LC-MS/MS) combined with multivariate statistical analysis was employed to screen for differential serum metabolites across groups. Metabolic pathway enrichment analysis of the selected differential markers was performed using the Human Metabolome Database (HMDB) and MetaboAnalyst 6.0. All processed S. nux-vomica seed products increased hepatic and renal coefficients and elevated hepatic and renal injury markers (AST, ALT, BUN, CRE), although these effects were reduced to varying degrees compared to the raw product. Overall, vinegar processing exhibited the most significant detoxification effect, followed by oil-fried>sand-roasted≈urine-soaked > raw seeds. Metabolomic analysis identified 14 differential metabolites across the groups, primarily including sphingolipids, glycerophospholipids, and vitamin-related compounds. Five significantly perturbed metabolic pathways were identified: sphingolipid metabolism, glycerophospholipid metabolism, α-linolenic acid metabolism, lipoic acid metabolism, and linoleic acid metabolism, with sphingolipid and glycerophospholipid metabolism common to all groups. In conclusion, processing of S. nux-vomica seeds reduced hepatorenal toxicity to varying degrees, and its toxic mechanisms may primarily involve disturbances in sphingolipid and glycerophospholipid metabolism.

The stem bark of Aquilaria yunnanensis (Thymelaeaceae) is traditionally used to treat cough and asthma in folk medicine. However, its active components have not been reported. In the present study, the chemical constituents of A. yunnanensis stem bark were isolated and identified, and their inhibitory activities against nitric oxide (NO) production in RAW264.7 cells induced by lipopolysaccharide (LPS) were evaluated to clarify the anti-inflammatory components in the stem bark of A. yunnanensis. A total of 22 compounds, including a new norlignan [(±)-1], were isolated from the ethanolic extract of A. yunnanensis stem bark by column chromatography, high-performance liquid chromatography (HPLC), recrystallization, etc. Through chiral separation of (±)-1, (-)-1 and (+)-1 were obtained. Based on electronic circular dichroism (ECD) calculations, the absolute configurations of (-)-1 and (+)-1 were determined to be (-)-(7R, 8S, 8′S)-4-O- (1, 3-dihydroxypropan-2-yl) zhebeiresinol and (+)-(7S, 8R, 8′R)-4-O- (1, 3-dihydroxypropan-2-yl) zhebeiresinol. The known compounds included (±)-zhebeiresinol (2), (+)-syringaresinol (3), (+)-episyringaresinol (4), orcinol (5), orsellinic acid (6), methyl orsellinate (7), 3, 4-dihydroxybenzoic acid (8), lecanoric acid (9), barbatic acid (10), 7β-hydroxy-β-sitosterol (11), β-sitosterol (12), stigmasterol (13), ergosterol peroxide (14), genkwanin (15), pilloin (16), 5-hydroxy-3′, 4′, 7-trimethoxyflavone (17), (-)-epicatechin (18), (-)-epicatechin gallate (19), tectorigenin 2-O-α-L-rhamnopyranoside (20), 2, 3-dihydroxypropyl docosanoate (21), and 1-O-linolenoylglycerol (22). According to the bioassay testing results, compounds 4 and 22 which inhibited NO production with IC₅₀ values of (32.82±1.04) and (18.50±0.95) μmol·L^-1, respectively, exhibited anti-inflammatory activities.

In this study, the total alkaloids of Corydalis delicatula were isolated and purified by a combination of silica gel, aluminum oxide basic column chromatography and other chromatographic techniques. The obtained compounds were structurally identified with the help of modern spectral resolution techniques such as nuclear magnetic resonance (1D/2D NMR), high-resolution mass spectrometry (HR-ESI-MS), and single crystal X-ray diffraction. Nine alkaloids were obtained from the total alkaloids of C. delicatula, encompassing two new compounds, corlicatine (1) and N-methyl-13-oxoprotopine (2), and seven known ones: (+)-ochotensine (3), rupestrine A (4), (+)-ochotensimine (5), oxohydrastinine (6), α-magnoflorine (7), 6, 7-methylenedioxy-N-methylisoquinoline (8), and N-methyl-6, 7-dimethoxyisoquinoline (9). The inhibitory activities of the compounds against acetylcholinesterase were evaluated, with compound 4 exhibiting moderate inhibition. Molecular docking was further performed to elucidate the binding mode of this compound. The results provide a basis for elucidating the medicinal substance basis of C. delicatula.

The chemical constituents from Sanguisorba officinalis were isolated and purified through macroporous resin, silica gel, polyamide, ODS, Sephadex LH-20 column chromatography, and preparative HPLC. Their structures were determined by extensive analysis of data from MS, NMR, OR, and calculated NMR and ECD. Twenty-six compounds (1-26) were isolated from the 95% ethanol extract of S. officinalis and identified as (1S, 4S, 5R, 7S, 10R, 11R)-1α, 11, 12-trihydroxy-eudesmane (1), vomifolilol (2), (3S, 5R, 6R, 7E)-3, 5, 6-trihydroxy-7-megastigmen-9-one (3), threo-1, 2-bis- (4-hydroxy-3-methoxyphenyl)-propane-1, 3-diol (4), erythro-1, 2-bis- (4-hydroxy-3-methoxyphenyl)-propane-1, 3-diol (5), phenylformic acid (6), vanillic acid (7), veratric acid (8), 3, 4-dimethoxy-5-hydroxybenzoic acid (9), 4- (1-hydroxy-1-methylethyl)-benzoic acid (10), p-hydrocoumaric acid (11), 4-hydroxyphenethyl alcohol (12), ω-hydroxypropioguaiacone (13), cytosporone V (14), indazole (15), 1H-indole-3-carboxylic acid (16), 7, 8-dimethyl-iso-alloxazine (17), (-)-jasmonic acid (18), 12-hydroxyjasmonic acid (19), (-)-methyl 12-hydroxyjasmonate (20), 5-hydroxyhexan-4-olide (21), (8R^*, 9R^*, 10S^*, 6Z)-trihydroxyoctadec-6-enoic acid (22), (9S, 10R, 11E, 13R)-9, 10, 13-trihydroxyoctadec-11-enoic acid (23), methyl (9S, 10R, 11E, 13R)-9, 10, 13-trihydroxyoctadec-11-enoate (24), pinellic acid (25), and (9E)-8, 11, 12-trihydroxyoctadecenoic acid methyl ester (26). Compound 1 was a new eudesmane-type sesquiterpenoid and named sanguisorbaol M. Compounds 3-5, 8, 10, 14-17, 19-23, 25, and 26 were isolated from Rosaceae for the first time. Compounds 2, 11, 12, 18, and 24 were firstly isolated from the genus Sanguisorba and compound 6 was isolated from S. officinalis for the first time. The inhibitory effects of the isolated compounds on melanin synthesis were evaluated via the 3-isobutyl-1-methylxanthine (IBMX)-induced B16F10 melanoma cell model. The results showed that compounds 1-3 at a concentration of 50 μmol·L^-1 had a significant anti-melanogenic activity.

This study established a hyperlipidemia model by feeding Sprague-Dawley rats a high-fat diet for 8 weeks. The rats were randomly assigned to the following groups: model group, atorvastatin calcium group (4.8 mg·kg^-1), low-, medium-, and high-dose Tanyu Tongzhi Optimization Decoction (TYTZD) groups (3.6, 7.2, and 14.4 g·kg^-1), and a normal diet control group. After 4 weeks of continuous administration, hematoxylin-eosin (HE) and oil red O staining were used to observe liver pathological changes and lipid infiltration. Automatic biochemical analyzer were performed to assess blood lipid profiles, coagulation function, and liver function. Transcriptomic and proteomic analyses were employed to identify differentially expressed genes (DEGs) and proteins (DEPs), followed by enrichment analysis. The MCODE algorithm was applied to classify DEGs and DEPs into modules, and network separation index (S_AB) was calculated to assess module separation, enabling construction of a gene-protein co-expression network for core target screening. The diagnostic accuracy of core targets was evaluated by area under the receiver operating characteristic (ROC) curve (AUC), and ELISA was used to measure core target expression. Western blot detected the expression of core pathway-related proteins in liver tissue. Results demonstrated that TYTZD significantly improved dyslipidemia, coagulation dysfunction, liver injury, hepatic pathology, and lipid infiltration in hyperlipidemic rats. Transcriptomic analysis identified 571 DEGs significantly reversed by TYTZD, mainly enriched in inflammatory signaling pathways such as Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB). Proteomic analysis identified 102 reversed DEPs, mainly involved in cholesterol metabolism pathways. Integrated analysis identified core targets including TLR4, tumor necrosis factor-α (TNF-α), integrin subunit alpha M (ITGAM), Toll-like receptor 2 (TLR2), matrix metalloproteinase 9 (MMP9), interleukin-1β (IL-1β), apolipoprotein E (APOE), and apolipoprotein C2 (APOC2), all with AUC values greater than 0.70. ELISA showed that TYTZD intervention significantly downregulated MMP9, TNF-α, IL-1β, TLR2, ITGAM, and TLR4, and upregulated APOC2 and APOE. Western blot indicated that TYTZD reduced TLR4, p-NF-κB, and IL-1β protein expression in liver tissue. In conclusion, TYTZD may exert anti-hyperlipidemic effects through regulation of core targets such as ITGAM, TLR4, and APOC2, and by modulating the TLR4/NF-κB signaling pathway to intervene in inflammatory responses and cholesterol metabolism, thereby achieving multi-target, multi-pathway therapeutic effects against hyperlipidemia.

This study investigated the asthma-alleviating effects of Poria cocos from the perspectives of immuno-inflammation and gut microbiota (GM), providing a theoretical basis for its rational clinical use. Male Balb/c mice were randomly divided into normal, model, positive control (montelukast sodium, 1.5 mg·kg^-1), low-dose P. cocos (0.057 g·kg^-1), medium-dose P. cocos (0.114 g·kg^-1), and high-dose P. cocos (0.227 g·kg^-1) group. Except for the normal group, the other groups were sensitized with ovalbumin (OVA) by intraperitoneal injection combined with nebulization to establish an asthma model, and treated with the corresponding drugs. After modeling, cough and wheezing indices, lung function, and asthma characteristic indicators including immunoglobulin E (IgE), interferon-γ (IFN-γ), interleukin 4 (IL4), and interleukin 10 (IL10) levels were measured. Lung tissue pathology was observed by hematoxylin-eosin (HE) and Masson staining. Quantitative real-time PCR (qRT-PCR) was used to detect airway inflammation-and remodeling-related mRNA levels in lung tissue. Levels of 12 inflammatory factors in serum and bronchoalveolar lavage fluid were detected by multiplex assay. Furthermore, the asthma-alleviating mechanism of P. cocos was analyzed by network pharmacology and GM 16S rRNA sequencing. Results showed that P. cocos reduced cough frequency in asthmatic mice, improved lung function, alleviated pathological damage in lung tissue, decreased serum IgE and IL 4 levels, increased IL 10 and IFN-γ levels, and multi-target regulated related inflammatory factors in serum and alveolar lavage fluid, maintaining immune homeostasis. Network pharmacology predicted that P. cocos exerts its asthma-alleviating effects via inflammation regulation pathways. GM sequencing showed that P. cocos effectively alleviated dysbiosis induced by OVA sensitization, significantly downregulated Muribaculum intestinale and Alistipes inops, and correlation analysis revealed both were positively correlated with pro-inflammatory factors. Therefore, it is speculated that P. cocos restores intestinal microecological homeostasis by reducing levels of M. intestinale and A. inops, decreases pro-inflammatory factor production, blocks the inflammatory cascade, and thereby exerts its asthma-alleviating effects.

This study investigates the inducing effects of jatrorrhizine (JAT), a bioactive compound extracted from Coptidis Rhizoma, on the p53-mediated ferroptosis in human colorectal cancer (CRC) cell lines HT29 and HCT116, specifically through the modulation of spermidine/spermine N1-acetyltransferase 1 (SAT1). Cell viability was assessed via the cell counting kit-8 (CCK-8) assay, while the colony formation assay was employed to determine the proliferative capacity. Cell cycle distribution was analyzed by flow cytometry, and mitochondrial membrane potential was evaluated via JC-1 dye. High-throughput RNA sequencing (RNA-seq) was performed to analyze transcriptomic changes in JAT-treated HT29 and HCT116 cells. Lentiviral transduction was utilized to generate SAT1-overexpressing HCT116 and HT29 cell lines, with transfection efficiency confirmed by qPCR. The levels of intracellular Fe²⁺, reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) were quantified via specific commercial assay kits. The protein and mRNA levels of key ferroptosis-related markers: solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4), tumor protein p53, and SAT1 were determined by Western blot and qPCR, respectively. The results demonstrated that JAT treatment significantly inhibited both cell viability and proliferation, induced cell cycle arrest, and reduced mitochondrial membrane potential in HCT116 and HT29 cells. RNA-seq results revealed that the KEGG pathways most significantly enriched in JAT-treated cells were protein digestion and absorption, arginine and proline metabolism, glycine, serine and threonine metabolism, and ferroptosis. Notably, the mRNA level of SAT1 in the arginine metabolism pathway was significantly upregulated following JAT treatment. Furthermore, JAT treatment increased the levels of Fe²⁺, ROS, and MDA, while markedly decreasing GSH content in a dose-dependent manner. In addition, the treatment downregulated the protein levels of the ferroptosis inhibitors SLC7A11 and GPX4, and upregulated the expression of the pro-ferroptotic proteins ACSL4, SAT1, and p53. SAT1 overexpression inhibited cell proliferation and activated the p53-mediated ferroptosis signaling pathway. The combination of SAT1 overexpression and JAT treatment (oveSAT1+JAT group) exhibited a synergistic effect, showing superior efficacy over SAT1 overexpression alone (oveSAT1 group). In conclusion, JAT activates the p53-dependent ferroptosis signaling pathway by upregulating the key metabolic enzyme SAT1, which is involved in proline metabolism, thereby inhibiting colorectal cancer cell proliferation.

This study investigated the effects and molecular mechanisms of sarsasapogenin (SSG) on colorectal cancer (CRC). After HRT-18 and HCT116 cells were treated with varying concentrations of SSG, the effect of SSG on CRC cell proliferation, migration, and glycolysis was assessed by cell counting kit-8 (CCK-8), real-time cellular analysis (RTCA), colony formation assay, high-content imaging analysis, and Seahorse glycolysis stress test. Changes in intracellular lactate release and adenosine triphosphate (ATP) levels under SSG treatment were measured by lactate and ATP detection kits, respectively. The effects of SSG on the transcription level and protein expression of the key glycolytic gene hypoxia-inducible factor 1α (HIF1α) were evaluated via quantitative polymerase chain reaction (qPCR) and Western blot. Direct binding between SSG and HIF1α was identified by drug affinity responsive target stability (DARTS). The UbiBrowser database, microscale thermophoresis (MST), and molecular docking simulations were employed to screen and validate the E3 ubiquitin ligase mediating HIF1α degradation. The results showed that SSG significantly inhibited CRC cell proliferation and colony formation in a dose-dependent manner. The 24-hour half-maximal inhibitory concentrations (IC₅₀) of SSG for HRT-18 and HCT116 cells were 6.275 μmol·L^-1 and 7.029 μmol·L^-1, respectively. After 24 hours of treatment with SSG at concentrations of 2.0, 4.0, and 6.0 μmol·L^-1, the HRT-18 and HCT116 clone formation rate was significantly reduced. High-content imaging results showed that SSG significantly inhibited the migration ability of CRC cells. Seahorse glycolysis stress test, along with lactate and ATP measurements, demonstrated that SSG suppressed basal glycolysis, glycolytic capacity, glycolytic reserve, lactate release, and ATP content in CRC cells. The qPCR and Western blot results showed that SSG had no significant effect on HIF1α mRNA expression, and the HIF1α protein expression was downregulated. DARTS confirmed the direct binding of SSG to HIF1α. UbiBrowser analysis identified five E3 ubiquitin ligases potentially targeting HIF1α. Molecular docking and MST results indicated that the F-box/WD repeat-containing protein 7 (FBXW7) exhibited strong binding affinity to HIF1α in the presence of SSG. In summary, SSG exerts its anti-cancer effects in CRC by targeting HIF1α to promote its FBXW7-mediated proteasomal degradation and consequently inhibiting glycolytic reprogramming.

This study explored the mechanism by which Suanzaoren Decoction (SZRD) ameliorates hippocampal neuronal damage in APP/PS1 mice by inhibiting Bax/Bcl-2/caspase-3-mediated apoptosis through the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Differentially expressed genes (DEGs) in Alzheimer's disease (AD) were screened using the GEO database, and KEGG and GO enrichment analyses were performed using R software. Thirty 6-month-old APP/PS1 transgenic mice were randomly assigned to three groups: APP/PS1 group, low-dose SZRD group (12.96 g·kg^-1), and high-dose SZRD group (25.92 g·kg^-1), while ten age-matched male C57BL/6J mice served as the blank control group. The treatment groups received SZRD by gavage for 28 consecutive days. Following treatment, spatial learning and memory were assessed using the Morris water maze test. Neuronal numbers were evaluated by Nissl staining. Immunohistochemistry (IHC) was used to detect neuron-specific nuclear protein (NeuN) and assess amyloid-β (Aβ) deposition in hippocampal neurons. Western blot was performed to measure the expression levels of PI3K/Akt pathway proteins and apoptosis-related markers, including Bcl-2, Bax, caspase-3, and cleaved-caspase-3 in hippocampal tissues. Bioinformatics analysis identified 686 DEGs, with GO enrichment linked to synaptic function and neurotransmitter transport, and KEGG pathways including neuroactive ligand-receptor interaction and PI3K/Akt signaling. In vivo experiments showed that, compared with APP/PS1 mice, both low-and high-dose SZRD groups exhibited improved learning and memory, increased neuronal numbers, and decreased Aβ deposition. Furthermore, hippocampal levels of p-PI3K/PI3K, p-Akt/Akt, and Bcl-2 were upregulated, while Bax and cleaved-caspase-3/caspase-3 levels were downregulated. In conclusion, SZRD may improve hippocampal neuronal damage and cognitive function in APP/PS1 mice by regulating Bax/Bcl-2/caspase-3-mediated apoptosis through the PI3K/Akt signaling pathway.

This article aims to investigate the inhibitory effect and molecular mechanism of sculponeatin A (STA) on triple negative breast cancer (TNBC). MDA-MB-436 and MDA-MB-468 were selected as cell models. The MTT assay, real-time cell analysis (RTCA), and colony formation assay were used to evaluate the effects of different concentrations of STA on the proliferation of TNBC cells. JC-1 staining and Annexin V-FITC/PI double staining combined with flow cytometry were used to measure the effect of STA on the apoptosis of TNBC cells. Western blot was employed to determine the expression changes of apoptosis-related proteins [cysteinyl aspartate-specific proteinase (caspase)-9, caspase-3, and poly-ADP-ribose polymerase (PARP)] and cancerous inhibitor of protein phosphatase 2A (CIP2A)/protein kinase B (AKT) signaling pathway proteins [CIP2A, AKT, phosphorylated (p)-AKT] in TNBC cells after STA treatment. To clarify the function of CIP2A in the action of STA, a CIP2A overexpression or CIP2A knockdown plasmid was transfected into cells, which were then treated with STA. Cell proliferation, apoptosis, and protein expression changes were evaluated by CCK-8, flow cytometry, and Western blot. Further, RT-qPCR and Western blot both showed that STA significantly downregulated the mRNA and protein levels of CIP2A and the protein level of c-Myc. The overexpression of c-Myc antagonized the downregulating effects of STA on the protein and mRNA levels of CIP2A, while the knockdown of c-Myc enhanced this effect. The drug affinity-responsive target stability (DARTS) assay and microscale thermophoresis (MST) assay confirmed the existence of direct binding between STA and c-Myc protein. The results indicated that STA significantly inhibited the proliferation and colony formation and induced the apoptosis of TNBC cells, manifested by an increased apoptosis rate, downregulated precursor protein expression of caspase-9 and caspase-3, and increased PARP cleavage. STA treatment reduced the p-AKT level but did not affect total AKT. Functionally, the knockdown of CIP2A enhanced the STA effects of inhibiting proliferation and inducing apoptosis, while the overexpression of CIP2A produced antagonistic effects. From a mechanism perspective, STA directly targets and binds to c-Myc to downregulate its expression, thereby inhibiting the transcription and translation of CIP2A and ultimately blocking the c-Myc/CIP2A signaling pathway. In conclusion, STA inhibits the malignant progression of TNBC by targeting the c-Myc/CIP2A signaling axis.