Cytotechnology最新文献

Viscum coloratum (Kom.) Nakai has been widely used in traditional Chinese medicine and clinical treatment mainly for rheumatoid arthritis (RA). Our latest study demonstrated that it could alleviate clinical symptoms of RA and decrease osteoclastogenesis-related gene expression in the CIA mouse model. However, whether Viscum coloratum (Kom.) Nakai could inhibit osteoclastogenesis and its underlying mechanisms need to be further elucidated. In this study, we demonstrated that extract of Viscum coloratum (Kom.) Nakai (EVC) exhibited extraordinary activity against RANKL-induced osteoclast differentiation identified by inhibiting the formation of TRAP-positive multinucleated osteoclasts, TRAP activity, and decreasing osteoclast-specific genes NFATc1 expression. The network pharmacology-based method integrating Protein-Protein interaction analysis revealed that Viscum coloratum (Kom.) Nakai could inhibit osteoclastogenesis by regulating PI3K-AKT signaling pathways. Then, the predicted signaling pathway of EVC on osteoclast differentiation was determined by Western blot. Using TRAP activity and nitric oxide assays, Viscumneoside III, which was identified as the main ingredient from Viscum coloratum (Kom.) Nakai in our previous study, was confirmed as the primary effective ingredient. These findings reveal that EVC executed the traditional effects on strengthening muscles and bones for the treatment of RA by inhibiting osteoclastogenesis via the regulated PI3K/AKT/mTOR signaling pathway, and Viscumneoside III was its effective ingredient.

Oxidative stress, inflammation, and impaired apoptotic signaling jointly contribute to glioma progression and treatment resistance. This study compared the effects of Trolox and melatonin on redox balance, inflammatory signaling, and apoptosis in U87-MG glioma cells using a comprehensive multi-parameter phenotypic panel, evaluated their potential complementary actions, and validated key findings in T98G cells. U87-MG cells were treated with Trolox (25-200 µM), melatonin (25-200 µM), or their combination (100 µM Trolox + 100 µM melatonin) for 6, 24, and 48 h. Cell viability, oxidative stress indices (total oxidant status, total antioxidant capacity, oxidative stress index), intracellular ROS and lipid peroxidation markers (MDA, 4-HNE), inflammatory cytokines (IL-6, TNF-α), apoptosis, and expression of Nrf2-HO-1-SOD2 and Bax/Bcl-2 were analyzed. NF-κB p65 activity and caspase-3/7 activation were assessed to clarify inflammatory and apoptotic signaling, with N-acetylcysteine used as a reference thiol antioxidant in selected assays. Melatonin induced dose-dependent antioxidant, anti-inflammatory, and pro-apoptotic effects at lower concentrations than Trolox, whereas Trolox primarily suppressed lipid peroxidation at higher doses. The combination treatment consistently produced the most pronounced effects across endpoints, including maximal oxidative stress reduction, NF-κB inhibition, and apoptosis induction. Similar response patterns were confirmed in T98G cells, while no cytotoxicity was observed in normal human astrocytes. These findings support dual redox modulation as a mechanistically rational adjuvant strategy in glioma research.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00915-x.

Microplastics (MPs) pose potential risks to human health due to their persistence and biotoxicity, mainly by inducing oxidative stress, inflammation, and gut microbiota imbalance. Exocarpium Citri Grandis (ECG), a traditional Chinese medicine with both medicinal and dietary value, has shown notable anti-inflammatory and antioxidant activities. In this study, UHPLC-Q-TOF MS profiling combined with network pharmacology analysis was applied to identify the principal effective ingredients of ECG and to evaluate its protective effects on polystyrene microplastic (PS) -induced hepatic and intestinal toxicity in mice. ECG and its main parts, naringenin and naringin, greatly reduced the oxidative stress caused by PS. They also eased liver injury and improved the intestinal barrier. ECG was associated with reduced inflammation and oxidative stress, accompanied by changes in the TLR4/NF-κB/NLRP3 and Nrf2/HO-1 pathways. It also improved intestinal structural integrity by increasing the levels of tight junction proteins. Gut microbiota analysis revealed that ECG intervention significantly improved PS-associated disruptions in microbial diversity and enriched beneficial genera such as Lactobacillus, Bacteroides, and Blautia. Metabolomic profiling showed that ECG significantly altered 1,507 PS-associated metabolic features, notably inhibiting pro-inflammatory arachidonic acid metabolism and upregulating antioxidant-related ubiquinone biosynthesis. Correlation analysis further linked key microbial shifts (e.g., Clostridia) with specific metabolite changes. In summary, ECG significantly alleviates PS-induced hepatoenterotoxicity through multi-level modulation of inflammatory, antioxidant, and gut microbiota-metabolite interactions, antioxidant, and gut microbiota-metabolism effects, providing new insights into the use of natural products to prevent and control health risks associated with MPs.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00921-z.

Metabolic dysfunction-associated fatty liver disease (MAFLD) is often asymptomatic in its early stages and exhibits a higher prevalence than both viral hepatitis and alcoholic liver disease, posing a significant challenge to modern healthcare. Qing-brick tea (QBT), a traditional Chinese tea characterized by its unique processing method and brick-like form, contains bioactive compounds including epicatechin, epicatechin gallate, gallic acid, and catechin.This study aims to investigate the preventive effects and underlying the mechanisms of QBT on MAFLD in mice induced by a high-fat diet in combination with dextran sulfate sodium in vivo and a hyperlipidemic environment of HepG2 cells in vitro. The results demonstrate that QBT effectively prevents the development of MAFLD, which may be associated with the activation of the AMPK/ACC pathway and the suppression of SREBP1/FAS signaling.These findings provide an experimental foundation and theoretical basis for the further development and utilization of QBT.

Objective: To investigate the effect of Ursolic acid (UA) on liver cancer cells through cell experiments, in order to inhibit cell proliferation and induce apoptosis.

Method: By adding z-vad-fmk apoptosis inhibitor and ROS inhibitor NAC, CCK8, cell apoptosis, Western blot experiments, mitochondrial membrane potential detection, and ROS flow cytometry were conducted. Compared with the positive drug sorafenib, the inhibitory effect of UA on liver cancer cells and its promotion of apoptosis were studied. The results of CCK8 confirmed that when UA was at its optimal concentration, its inhibitory effect on liver cancer cell viability was greater than that of sorafenib; Through cell apoptosis experiments, it can be observed that under fluorescence microscopy, the fluorescence intensity in the UA+HepG2 group is significantly stronger than that in the sorafenib+HepG2 group; Western blot experiments showed that the relative expression levels of proteins in HepG2 were almost equal between the HepG2+UA and sorafenib+HepG2 groups; through ROS experiments, it was found that when the reactive oxygen species in the HepG2+UA group were lower than those in the sorafenib+HepG2 group, it indicated that the former had a stronger effect of UA on HepG2.

Conclusion: UA has a significant inhibitory effect on proliferation and promotes apoptosis in liver cancer cells.

Across multiple aspects, bamboo is intricately associated with human civilization and is well known for its numerous uses. The shoots of bamboo are consumed all over the world, and for its taste, fragrance, and nutritional value, it has been treated as a delicacy. Phyllostachys pubescens is a major bamboo species in Japan. Although bamboo shoots have been studied for various biological activities, research evaluating their skin whitening and anti-aging properties is limited. This study evaluated bamboo shoots for anti-melanogenesis properties. Using a bio-guided isolation approach, one active compound was identified: L-(+)-Lactic acid (2-hydroxypropanoic acid; 2-HPA). The structure of the isolated compound has been elucidated using 1D-Nuclear magnetic resonance (NMR) as well as mass spectrometry (MS) analysis and comparison with previously published literatures. 2-HPA exhibited dose-dependent downregulation in melanin production and tyrosinase inhibition. Comparative data of the stereoisomers showed, chirality influences anti-melanogenesis activity, while the L stereoisomer showed superior activity. 2-HPA could also downregulate the expression of the tyrosinase gene dose dependently. 2-HPA has showed anti-aging potential via upregulation of the production of collagen and hyaluronic acid in normal human dermal fibroblast (NHDF-Ad) cells and modest downregulation of elastase activity. 2-HPA was found to provide supportive protection of fibroblast cells against H2O2-induced oxidative stress. These preliminary data suggest bamboo shoot to be a potent tyrosinase inhibtor to downregulate the melanin production, therefore to be utilized in the cosmeceutical and other health related industries.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00907-x.

Lentiviral vectors (LVs) have revolutionized gene therapy by enabling stable gene integration into dividing and non-dividing cells, addressing critical challenges in treating genetic disorders. The transition from second to third-generation LVs has increased biosafety by minimizing the risk of replication-competent lentiviruses and expanded their clinical applicability. Despite significant advancements, producing high-titer functional LVs, particularly at an industrial scale, remains a considerable challenge due to the need for enhanced scalability, cost-efficiency, and effectiveness. This Review delves into cutting-edge innovations in LV production, from optimized transient transfection in various cell lines to the development of stable producer cell lines. Stable producer cell lines offer unparalleled scalability but face challenges related to viral protein cytotoxicity. Inducible systems have emerged as pivotal tools for addressing these problems, allowing for precise gene expression and controlled production. Additionally, advancements in bioprocess engineering, ranging from optimized culture conditions, including pH and media composition, to novel bioreactor technologies like structured fixed-bed systems, continue to redefine industrial-scale LV production. These breakthroughs, coupled with the analysis of costs and efficiencies of various methodologies, can further illustrate the potential for large-scale LV production and facilitate widespread therapeutic applications.

Background: Capparis spinosa L., rich in flavonoids and polyphenols, has limited mechanistic evidence regarding its anticancer effects. This study evaluated the methanol extract of C. spinosa in PC3 prostate cancer cells using combined in vitro and in silico analyses.

Methods: Cytotoxicity in PC3 and HUVEC cells was assessed by MTT, followed by qRT-PCR and Western blot analysis of Bcl-2/Bax. Colony formation and wound-healing assays evaluated functional effects. Kaempferol and quercetin were analyzed via SwissADME, admetSAR, PASS Online and docking against apoptosis-related proteins.

Results: The extract showed selective cytotoxicity toward PC3 cells (IC₅₀: 12.4 mg/mL at 24 h; 10.6 mg/mL at 48 h) with minimal HUVEC toxicity. Treatment suppressed Bcl-2 and increased Bax, reduced colony formation (~ 70%), and inhibited migration. Docking showed strong binding of kaempferol and quercetin to Bcl-2 (≈ - 8.1 kcal/mol).

Conclusion: C. spinosa extract induces mitochondrial apoptosis and inhibits PC3 proliferation, supporting its potential as a phytotherapeutic candidate.

Breast cancer stem cells (BCSCs) are one of the causes of drug resistance and disease recurrence due to their capacity for self-renewal and heterogeneity induction. A new BCSC-targeting agent has become a prospective approach to overcome resistance. Baicalein (5,6,7-trihydroxyflavone), a flavonoid extracted from Scutellaria baicalensis, has demonstrated anticancer activities in several models, including breast cancer. However, further elucidation of its effects on BCSCs is required. This study utilizes integrative bioinformatic approaches to identify the potential targets of baicalein in overcoming BCSC. In vitro experiments confirmed the top ten target genes recognized during a previous bioinformatic analysis of MCF-7 as a mammosphere for cytotoxicity and gene expression assays. We identified the potential baicalein target genes in BCSCs, which include CTNNB1, STAT3, BCL2, HIF1A, ESR1, TNF, CCND1, IL6, JUN, and MAPK3. Gene annotation and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway analysis revealed a possible attenuation of the estrogen signaling pathway by baicalein and its involvement in cell cycle regulation. We successfully constructed a three-dimensional (3D) mammosphere culture and characterized it to possess higher levels of "stemness-associated" factors (OCT4 and SOX2) compared to two-dimensional (2D) cultures. Baicalein did not demonstrate any significant effects on cell viability in both 2D and 3D cultures, although a decline was observed in 2D cultures. qRT-PCR revealed that baicalein suppressed all hub genes. Furthermore, molecular docking confirmed the gene expression patterns and that baicalein had better binding affinity to CTNNB1, STAT3, TNF, JNK1, and mitogen-activated protein kinase (MAPK) than the respective native ligands. In addition, other proteins also interacted with baicalein, as reflected by the docking scores. Baicalein was identified to interact with ten potential targets through a bioinformatics study, although it did not exhibit cytotoxicity in 2D and 3D MCF-7 cultures. However, a downward trajectory was observed in the expression levels of hub genes related to kinase pathways, like Wnt/β catenin, PI3K/Akt, and MAPK, as well as inflammation-associated genes that correlate with BCSC survivability. One of the most prominent was the estrogen signaling pathway, which was supported by the molecular docking results. Future research directions included confirmation of baicalein's efficacy and toxicity through in vivo approaches, as well as to understand its efficacy as a combination chemotherapeutic agent.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00917-9.

Trans- ferulic acid (TFA) is a byproduct of hydroxyl cinnamic acid and it has been reported that TFA possesses different biological activities such as an anti-inflammatory, anti-cancer, anti-antioxidant, and antibacterial. Due to its weak solubility and insufficient absorption in gastrointestinal tract, use of trans- ferulic acid in treating a variety of disorders is constrained. Thus, to address these issues, chitosan and tragacanth bicomplex were used to encapsulate TFA for proper delivery. Additionally, the anticarcinogenicity and in-vivo anti-inflammatory potential of TFA-loaded tragacanth-chitosan nanoparticles were investigated. Anticarcinogenic potential of the synthesized nanoparticles were evaluated by using vero cell line was as non-cancerous cells and Hela as well as MBMB cell line as cancerous cell and it was observed that trans-ferulic acid loaded nanoparticles are non-cytotoxic under the tested conditions, as over 80% of vero cells remained alive after 24 h at the lowest concentration. Cytotoxicity of these nanoparticles on HeLa and MDA-MB-231 cells showed concentration-dependent cytotoxicity after 48 h, with 86.68% cell death observed in Hela cells and 94.54% in MDA-MB-231 cells at 60µM concentration. Carrageenan-induced rat paw edema was used to evaluate the in-vivo anti-inflammatory activity. It was observed that TFA loaded nanoparticles decreased rat paw edema by 40.67% and increased cell viability by 84.03 ± 0.62% in vero cell lines. A statistically significant data was considered if p-value less than 0.05. This work suggest that encapsulated trans-ferulic acid be more suitable for use in various clinical applications.