Pharmaceutical Biology最新文献

Context: Rheum palmatum L. (Rhubarb) has shown potential in managing Chronic Kidney Disease (CKD) but its protective mechanisms remain unclear.

Objective: This study investigates rhubarb's therapeutic effects and underlying mechanisms in CKD.

Materials and methods: High-performance liquid chromatography (HPLC) established a rhubarb fingerprint to ensure quality control. Network pharmacology and Mendelian randomization identified primary CKD therapeutic targets. Inflammation, oxidative stress, and renal performance were assessed through ELISAs and biochemical tests. Renal structure and fibrosis were examined using hematoxylin-eosin and Masson staining. Protein expression related to fibrosis, apoptosis, and NF-κB pathway activity was measured via Western blotting. Discovery Studio 2019 (DS 2019) was used for molecular docking.

Results: HPLC fingerprinting confirmed high batch-to-batch consistency of rhubarb, identifying five key anthraquinones (aloe-emodin, rhein, emodin, chrysophanol, physcion) with similarity indices >0.91. Network pharmacology identified 19 active compounds targeting 2,597 CKD-related proteins, with 47 overlapping targets including IL6, TNF, TP53, CASP3, and IL1B as core nodes. MR analysis demonstrated a statistically significant causal association between TNF and CKD (OR = 1.02, p < 0.05), with positive trends for IL6 and CASP3. In CKD rat models, rhubarb significantly improved renal function by reducing blood urea nitrogen (BUN), serum creatinine (SCr), and uric acid (UA) levels (p < 0.05). Histopathology showed reduced glomerulosclerosis, tubular atrophy, interstitial fibrosis, and inflammatory infiltration after treatment. Rhubarb markedly decreased renal fibrosis markers including collagen I, collagen III, α-SMA, and TGF-β (p < 0.01). Pro-inflammatory cytokines IL-6, IL-1β, and TNF-α levels were significantly suppressed (p < 0.001). Antioxidant enzyme activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were restored, while lipid peroxidation (LPO) was reduced (p < 0.05). Rhubarb inhibited NF-κB pathway activation by lowering phosphorylated NF-κB and IκBα, and increasing total IκBα expression (p < 0.01). Apoptosis-related proteins showed upregulated Bcl-2 and downregulated Bax and cleaved caspase-3 expression (p < 0.05). Molecular docking confirmed strong binding affinities of rhubarb's core compounds with key targets such as TNF and IL6, supporting their therapeutic roles.

Conclusion: Rhubarb significantly reduces renal impairment, fibrosis, inflammation, and apoptosis through the NF-κB pathway, supporting its traditional use and potential as an adjunct therapy for CKD.

Context: Ultraviolet B (UVB) radiation is a key environmental contributor to skin photoaging, primarily by inducing oxidative stress, mitochondrial dysfunction, metabolic imbalance, and downregulation of tight junction (TJ) proteins. Cedrol, the major component of the essential oil from Cunninghamia lanceolata var. konishii, a tree species endemic to Taiwan, exhibits antioxidant properties. However, its restorative effects against UVB-induced skin damage have not been fully elucidated.

Objective: In this study, HaCaT keratinocytes were used to evaluate the post-treatment effects of cedrol on UVB-induced damage to skin cells.

Materials and methods: HaCaT cells were exposed to UVB irradiation followed by cedrol treatment. Cell viability, intracellular reactive oxygen species (ROS), mitochondrial membrane potential, ATP levels, mitochondrial biogenesis-related proteins (SIRT1, PGC-1α, Nrf2, TFAM), and TJ proteins (ZO-1, occludin, claudin-3) were assessed. Additionally, ¹H-NMR-based metabolomics was conducted to evaluate UVB-induced metabolic changes.

Results: Cedrol significantly improved cell viability post-UVB exposure, decreased intracellular reactive oxygen species (ROS), and restored mitochondrial membrane potential and ATP levels. It also upregulated mitochondrial biogenesis-related proteins (SIRT1, PGC-1α, Nrf2, and TFAM) and maintained TJ protein expression (ZO-1, occludin, and claudin-3), thereby preserving epithelial barrier integrity. Furthermore, ¹H-NMR-based metabolomics revealed that cedrol mitigated UVB-induced metabolic disturbances, particularly in amino acid and energy pathways.

Discussion and conclusion: Cedrol alleviates UVB-induced cellular damage by modulating mitochondrial function and metabolic homeostasis, indicating its potential as a natural agent for promoting skin recovery after UV exposure.

Context: Recent research has revealed significant advancements in the field of traditional Chinese medicine (TCM) for skin diseases. However, there is a lack of visualization analysis within this research domain.

Objective: To analyze the research directions and advancements in TCM research in skin diseases.

Materials and methods: Publications related to TCM in skin diseases from 2014 to 2024 were searched on the Web of Science Core Collection (WoSCC), VOSviewer, CiteSpace, and the R package "bibliometrix" were employed to visualize and analyze the retrieved data.

Results: The study included 527 articles published in 25 countries. The number of publications consistently increased from 2014 to 2024. The Guangzhou University of Chinese Medicine was the most noteworthy institution in this field. Among the journals in this domain, the Journal of Ethnopharmacology was the most popular, and most frequently co-cited journal. Chuanjian Lu published the most papers and Yin-Ku Lin was the most frequently co-cited author. Among keywords, "psoriasis" appeared the most frequently. Additionally, several emerging research hotspots were identified, indicating the transition from traditional Chinese therapies to investigations of the molecular interactions and network pharmacology of Chinese herbs in treatment of skin diseases over the past decade.

Discussion and conclusion: This visualization analysis summarizes the research directions and advancements in TCM research on skin diseases. It presents a comprehensive examination of the latest research frontiers and trends and serves as a valuable reference for scholars engaged in the study of TCM research.

Background: Apium graveolens L. (celery) is a dietary vegetable with anti-inflammatory properties. It has the potential to treat acute lung injury (ALI) caused by COVID-19 or other diseases.

Objective: To investigate the effects of Apium graveolens water extract (AGWE) on ALI in human lung A-549 cells induced by lipopolysaccharide (LPS).

Materials and methods: A-549 cells were treated with AGWE for 24 h and then stimulated with 10 μg/mL LPS for another 24 h. The effects of AGWE on cell viability, the inflammatory response, oxidative stress, and apoptosis and their regulatory factors, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and NLR family pyrin domain containing 3 (NLRP3) inflammasome signaling activation were analyzed.

Results: Treatment with 5-50 μg/mL AGWE reversed the decrease in cell viability caused by LPS (p < 0.05). AGWE can reduce interleukin (IL)-1β, IL-6, IL-18, and TNF-α levels; their EC₅₀ values are 61.4, 65.7, 37.8, and 79.7 μg/mL, respectively. AGWE can reduce reactive oxygen species and thiobarbituric acid reactive substances in A-549 cells induced by LPS. AGWE also reduced the levels of apoptosis (EC50 of 74.8 μg/mL) and its regulators (Bid; Caspase-9, -8, and -3; Bax) and increased the levels of the mitochondrial membrane potential in A-549 cells induced by LPS. AGWE can also decrease the protein levels of NLRP3 and Caspase-1 and the activation of NF-κB signaling in A-549 cells induced by LPS.

Conclusions: These results show that 10 and 50 μg/mL AGWE can reduce the acute inflammation induced by LPS by reducing NF-κB and NLRP3 inflammasome signaling and mitochondria-dependent apoptosis pathways.

Context: Total flavonoids from Litchi chinensis Sonn. (Sapindaceae) seeds (TFLS) effectively attenuate stem cell-like properties in breast cancer cells. However, their pharmacological effects and mechanisms in suppressing breast cancer metastasis remain unclear.

Objective: This study aimed to elucidate the inhibitory effects and underlying mechanisms of TFLS on breast cancer metastasis.

Materials and methods: The antiproliferative, migratory, and invasive activities of breast cancer cells following TFLS treatment were evaluated using CCK-8, wound-healing, and transwell assays. The epithelial-mesenchymal transition (EMT) biomarkers were evaluated via Western blot analysis. The anti-metastatic effects of TFLS were further validated in vivo using zebrafish and mouse models. Network pharmacology methodology was utilized to predict potential targets and signaling pathways, which were subsequently corroborated by Western blot. Potential active compounds were identified through molecular docking, and the chemical constituents of TFLS were analyzed and characterized using UPLC-QTOF/MS.

Results: TFLS suppressed the proliferation of MDA-MB-231 and MDA-MB-468 cells, with IC₅₀ values of 44.47 μg/mL and 37.35 μg/mL at 72 h, respectively. It effectively suppressed breast cancer metastasis in vitro, demonstrated by a marked reduction in cellular motility and invasiveness, alongside the reversal of EMT. Consistent with pathway enrichment analysis, network pharmacology revealed that TFLS reduced the phosphorylation levels of PI3K, AKT, mTOR, JNK, ERK, and p38 in breast cancer cells. Molecular docking identified seven potential active ingredients, and UPLC-MS/MS confirmed the presence of key compounds, including procyanidin A2.

Discussion and conclusion: TFLS effectively inhibits breast cancer cell proliferation, migration, and invasion in vitro by reversing the EMT phenotype, while suppressing metastasis in vivo. These effects are likely mediated via the attenuation of the PI3K/AKT/mTOR and MAPK signaling pathways.

Context: Lipid nanoparticles (LNPs) are the primary non-viral vectors for siRNA delivery. However, synthetic lipids face issues, such as low lysosomal escape efficiency and high cost.

Objective: This study aimed to use three natural lipids to construct LNPs, optimize their preparation and freeze-drying processes, and evaluate their siRNA delivery efficiency in vitro.

Materials and methods: Coix seed lipid [Coix lacryma-jobi L. var. mayuen (Roman.) Stapf (Poaceae), CSL], Brucea javanica seed lipid [Brucea javanica (L.) Merr. (Simaroubaceae), BJL], and Soybean oil [Glycine max (L.) Merr. (Fabaceae), SO] were used to construct LNPs. The Z-average size, zeta potential, Polymer Dispersity Index, and N/P ratio of the LNPs were characterized. Transmission electron microscope was used for morphology observation and the MTS assay for cytotoxicity. Confocal laser scanning microscope assessed cell uptake, lysosomal escape, and co-localization of lipid droplets. The efficiency of siRNA knockdown was evaluated in three cells using qPCR and Western blot. The freeze-drying processes were optimized.

Results: The optimal LNPs exhibited a size of 160-180 nm, zeta of 44-50 mV, and PDI of <0.2. At 200 μg/mL, the LNPs did not affect cell viability. CSL-LNPs, BJL-LNPs, and SO-LNPs reduced KRAS^G12D mRNA levels in AsPC-1 cells by 67.87 ± 3.89, 47.18 ± 7.65, and 42.52 ± 8.90%, respectively. Freeze-dried LNPs retained their basic physical properties and the three LNPs reducing KRAS^G12D mRNA levels by 58.47 ± 4.00, 51.83 ± 4.57, and 38.00 ± 4.89%, respectively.

Discussion and conclusion: Natural lipids are promising components for LNPs construction, offering new avenues for siRNA delivery in gene therapy.

Context: Matrine has antinociceptive properties, and spinal cord ionomic changes are involved in bone cancer pain.

Objective: To investigate the relationship between ionomic metabolism in cerebrospinal fluid (CSF) and spinal cord and matrine's analgesic efficacy.

Materials and methods: The antinociceptive effects of matrine were identified in rats via intraperitoneal (i.p.) injection using the tail-immersion and formalin tests. Pharmacodynamic parameters for matrine against formalin-induced pain were calculated with nonlinear regression analysis. Inductively coupled plasma mass spectrometry (ICP-MS) technology was utilized to detect contents of the ionome in CSF and spinal cord. Variations in ionomic metabolism in different treated groups were examined using Pearson's correlation coefficients and principal component analysis (PCA).

Results: In the tail-immersion test, matrine significantly prolonged tail-flick latency in rats. Matrine also dose-dependently yielded analgesia against formalin-induced biphasic pain, with an onset at around 10 min post-injection and a duration of 100 min. The ED₅₀ and E_max values were 19.01 mg/kg and 71.86% for phase I and 40.30 mg/kg and 81.51% for phase II, respectively. Pearson's correlation coefficient study and PCA revealed significant reprogramming of ionomic metabolism in the CSF and the spinal cord in the NM (normal saline + matrine), NF (normal saline + formalin), and FM (formalin + matrine) groups, compared to the NN (normal saline + normal saline) group.

Discussion and conclusions: These findings broaden the known analgesic spectrum of matrine and provide novel insights into the involvement of ionomic metabolism in its analgesic efficacy.

Context: Quinoline, isoxazole, and pyridothiazolopyrimidinone derivatives are novel compounds with significant biological activity, exhibiting anticancer properties and holding promising therapeutic applications.

Objective: This investigation synthesized new heterocyclic compounds in high yields from quinoline-2-thioxo-pyridopyrimidinone and assessed their anticancer activities. Additionally, it conducted molecular docking, ADMET analysis, and molecular dynamics simulations.

Materials and methods: A new series of quinoline-pyridothiazolopyrimidine derivatives has been synthesized using advanced techniques. The structures of the new compounds were confirmed using IR, NMR, MS and elemental analysis. All compounds were tested in vitro for their anticancer activity.

Results: Isoxazole and thiazolopyridopyrimidinones displayed the highest activity against several cancer cell lines. Docking simulations revealed that compounds 5d, 5e, 11a, and 11b exhibited favorable binding energies and effectively interacted with the active sites of the EGFR, CDK2, ERα, and VEGFR receptors. The ADMET analysis of these compounds demonstrated compliance with Pfizer's rules. Molecular dynamics simulations confirmed the stability of complexes formed by compounds 5d, 11a, and 11b with CDK2, ERα, VEGFR, and EGFR. The root mean square deviation (RMSD) values were recorded, while the root mean square fluctuation (RMSF) values ranged from 0.10 to 0.6 nm. The solvent-accessible surface area (SASA) values were measured to be between 135-145 nm², 125-130 nm², 155-165 nm², and 160-175 nm².

Discussion and conclusions: The cytotoxicity (IC₅₀) and selectivity index are presented in Tables. Molecular docking analyses showed that compounds 5d, 5e, 11a, and 11b demonstrated significant binding energies. These consistent results support the notion that both practical and theoretical studies align regarding the anticancer properties of these new compounds. Furthermore, these findings emphasize the potential of these compounds in ongoing drug development efforts.

Context: Breast and liver cancers remain significant global health challenges, necessitating the discovery of novel anticancer agents. Marine cyanobacteria, such as Moorena producens belonging to the family: Oscillatoriaceae, are rich sources of bioactive compounds with potential anticancer properties.

Objective: This study aims to identify and characterize bioactive compounds from M. producens and evaluate their anticancer activity against breast and liver cancer cell lines.

Materials and methods: M. producens was collected and authenticated using 16S rRNA sequencing. The ethanolic extract was analyzed using LC-QTOF-MS/MS to identify the potential bioactive metabolites. Network pharmacology analysis was employed to predict the potential targets of these compounds. The crude extract was fractionated, and the fractions were screened for the anticancer activity against MCF-7 and HepG2 cell lines.

Results: LC-QTOF-MS/MS analysis identified 25 metabolites, including apocarotenoids, spirovetivane alkaloids, and toxins. Network pharmacology analysis suggested that malyngamide D, isomalyngamide I, mueggelone, 11,12-didehydrospironostoic acid, and 12-hydroxy-2-oxo-11-epi-hinesol were potential bioactive compounds targeting proto-oncogene tyrosine-protein kinase (Src), mitogen-activated protein kinase 3(MAPK3), and MAPK1 kinases. Molecular docking studies further supported these findings, with 11,12-didehydrospironostoic acid exhibiting strong binding affinities to Src and MAPK1 kinases. Among the nine fractions obtained, Fraction 1 showed the most potent anticancer activity against both MCF-7 and HepG2 cell lines, with IC₅₀ values, 59.63 ± 7.1 and 149.23 ± 0.9 µg/mL, respectively.

Discussion and conclusion: The results of this study highlight the potential of M. producens as a source of novel anticancer compounds. Further investigation of the bioactive compounds in Fractions 1 and 2 may lead to the discovery of promising anticancer agents.

Context: cAMP-induced intestinal chloride secretion plays a pivotal role in the pathogenesis of secretory diarrheas.

Objective: In this study, we investigated the antisecretory effects of α,β-dehydromonacolin K, a derivative of lovastatin from Aspergillus sclerotiorum, on cAMP-induced chloride secretion in human T84 cells and fluid secretion in human colonoids.

Materials and methods: Short-circuit current analyses and swelling assays were used to investigate the effects of α,β-dehydromonacolin K on chloride transport and fluid secretion, respectively. Proteomic analyses were performed to determine the potential anti-diarrheal mechanisms of α,β-dehydromonacolin K.

Results: In T84 cells, α,β-dehydromonacolin K inhibited cAMP-induced chloride secretion with an IC₅₀ of ∼ 6.32 μM. Apical chloride current analyses demonstrated that α,β-dehydromonacolin K inhibited CFTR chloride channels stimulated by cAMP agonists with an IC₅₀ of ∼ 1 μM. Basolateral potassium current analyses indicated that α,β-dehydromonacolin K had no effect on basolateral potassium channel activities. In a three-dimensional (3D) model of human colonoids, α,β-dehydromonacolin K (20 µM) suppressed both cAMP-induced and calcium-induced fluid secretion by ∼ 70%. Proteomic analyses of human colonoids revealed that α,β-dehydromonacolin K interacted with 33 proteins, including those associated with non-sense-mediated mRNA decay (NMD). Notably, the inhibitory effects of α,β-dehydromonacolin K on cAMP-induced chloride and fluid secretion were significantly diminished in the presence of SMG1i, an inhibitor of serine/threonine-protein kinase SMG1 involved in NMD, suggesting that α,β-dehydromonacolin K inhibits cAMP-induced chloride-driven fluid secretion in human intestinal epithelial cells by mechanisms involving SMG1-dependent NMD pathways.

Discussion and conclusions: α, β-Dehydromonacolin K represents a promising class of natural compounds that exert antisecretory effects in human intestinal epithelia via a novel mechanism of action involving SMG1 in NMD pathways.