Pasarapa Towiwat

Mitragynine, the principal alkaloid of Mitragyna speciosa (kratom), exhibits opioid-like analgesic effects but is associated with tolerance following prolonged exposure. Schwann cells are particularly vulnerable to chemically induced toxicity, and disruption of their homeostatic functions has been implicated in neurotoxic injury. The cellular mechanisms underlying this adaptive response remain poorly understood. In this study, an integrated quantitative proteomics and systems biology approach was performed to investigate mitragynine-induced molecular remodeling in RSC96 Schwann cells. Cells were exposed to 20 µM mitragynine, the highest non-cytotoxic concentration, for 72 h and analyzed using liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based proteomic profiling. Differentially expressed proteins were characterized through Gene Ontology enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Reactome pathway mapping, InterPro domain annotation, and protein–protein interaction network reconstruction using STRING and BioGRID. Proteomic profiling identified 91 significantly altered proteins, comprising 60 downregulated and 31 upregulated proteins. Functional enrichment revealed coordinated suppression of translational machinery, cytoskeletal organization, and metabolic pathways associated with Schwann cell homeostasis. Network reconstruction highlighted AMP-activated protein kinase (AMPK) as a high-centrality node within the downregulated interaction network. Upregulated proteins were enriched in xenobiotic stress responses, aminoacyl-tRNA biosynthesis, and chromatin remodeling pathways. Structural similarity analysis revealed limited overlap between the mitragynine scaffold and morphine despite their shared receptor target. These findings suggest that chronic mitragynine exposure induces coordinated proteomic and network-level remodeling in Schwann cells, identifying regulatory pathways consistent with tolerance-related cellular adaptation and peripheral neurotoxic risk.

Thunbergia laurifolia Lindl. is a plant known for its promising biological activity, including antioxidant and anti-inflammatory activities, and a rich source of rosmarinic acid (RA). The extraction of T. laurifolia for cosmetic and skincare products using conventional solvents has encountered difficulties, including safety concerns, skin irritation, undesirable odors, and inefficient extraction. In this work, 14 types of natural deep eutectic solvents (NaDESs) with varying compositions and ratios were investigated to compare their efficiency in extracting RA from T. laurifolia by HPLC analysis. The NaDES with the highest extraction efficiency was further utilized in ultrasonic-assisted extraction (UAE), and the extraction parameters were optimized using response surface methodology. The optimized RA content and DPPH scavenging activity were predicted by response surfaces methodology to be 7.52 mg/g DW and 37.6 mg TE/g DW, respectively. The optimal extraction condition was achieved using a propylene glycol-lactic acid NaDES (at an 8:1 molar ratio) with 37% (w/w) H₂O, a 30 mL/g liquid-to-solid ratio, an 80 °C extraction temperature, and a 32 min extraction time. The optimized extract was proved to suppress ROS in H₂O₂-induced keratinocytes. The extract demonstrated robust stability against basic, oxidative, and photolytic stresses, and maintained long-term chemical stability up to 90 days. This study introduces a new green solvent for the effective extraction of T. laurifolia, thereby improving the safety and quality of the extracts for skincare and cosmetic products.

Selective α7 nicotinic acetylcholine receptors (α7 nAChRs) have emerged as therapeutic targets for managing various pain conditions, including inflammatory pain. Activation of α7 nAChRs by selective agonists has been shown to exert both antinociceptive and anti-inflammatory effects. In this study, we investigate the pharmacological effects of 3-(4-Hydroxyphenyl-1,2,3-triazol-1-yl)-quinuclidine (QND8), a selective and potent α7 nAChR agonist, in a murine model of carrageenan-induced inflammatory pain and evaluated its central nervous system (CNS) safety profile. QND8 significantly alleviated thermal and mechanical hyperalgesia at doses of 1, 3, and 10 mg/kg, and reduced carrageenan-induced paw edema at doses of 10 mg/kg. Importantly, QND8 exhibited these effects without impairing motor coordination and general behaviors, as demonstrated by the rotarod test and automated home cage behavioral analysis. These findings highlight QND8 as a promising preclinical analgesic candidate with anti-inflammatory efficacy and a favorable CNS safety profile. Computational modeling further revealed a stable and high-affinity binding of QND8 within the orthosteric site of α7 nAChR, consistent with its selective agonist activity. The results support further development of QND8 as a selective α7 nAChR-targeting therapeutic for inflammatory pain.

Sickness behaviors are a natural response to inflammation and pathogenic shocks. However, current treatments have limitations, highlighting the need for novel anti-inflammatory agents. Aloifol I, which is a dihydrostilbenoid, was investigated as a potential candidate. Its anti-inflammatory properties were first evaluated in vitro using lipopolysaccharide (LPS)-induced RAW 264.7 macrophage and BV-2 microglial cells, followed by proteomic analysis to elucidate underlying mechanisms. In vivo efficacy was then evaluated in a mouse model of LPS-induced sickness behaviors at 12.5, 25, and 50 mg/kg doses. Central nervous system (CNS) safety was evaluated at 50 mg/kg by assessing the general behavior and motor coordination of mice. Results demonstrated that aloifol I significantly suppressed the LPS-induced IL-6 and TNF-α release in both macrophage and microglia. Proteomic analysis revealed that aloifol I downregulated proteins involved in translation, glycolysis, and cytoskeletal organization while upregulating proteins related to mitochondrial function, stress response, and inflammation resolution, suggesting its multifaceted anti-inflammatory mechanism. In vivo, aloifol I attenuated LPS-induced fever from 38.3 °C to the basal temperature of 36 °C, confirming its antipyretic effect. It also improved LPS-induced locomotor impairments in a dose-dependent manner, reflecting its ability to alleviate inflammation-associated behavioral impairments. Additionally, aloifol I significantly reduced LPS-induced pro-inflammatory cytokine levels of IL-6 and TNF-α in both plasma and brain tissues, suggesting peripheral and central anti-inflammatory effects. Importantly, no adverse effects on motor coordination or general behaviors were observed, supporting a favorable CNS safety profile. These findings collectively highlight the promising therapeutic potential of aloifol I as a potential anti-inflammatory agent for the treatment of inflammation-related sickness conditions.

The prevalence of alcohol consumption among the younger generation remains alarmingly high. A hangover is a common short-term consequence observed after consuming alcohol. To effectively study alcohol-induced hangovers, reliable and translational animal models, along with appropriate testing methods, are required. While several testing approaches have been used in hangover-induced mice, they often fail to assess innate behaviors comprehensively and are limited by short observation periods. Although existing studies have developed methods to assess hangover-related behaviors in rodents, few have focused on innate behaviors. This study aimed to establish a model for assessing the innate behaviors of hangover-induced mice using automated home-cage-like behavioral monitoring. Mice were intraperitoneally injected with ethanol at doses of 3, 2, or 1 g/kg, followed by behavioral assessments, including exploratory actions and long-term home-cage-like behaviors during both day and night phases. Results showed a significant reduction in mobile behaviors (climbing, locomotion, rearing), speed, and distance traveled, along with increased immobility in both exploratory and long-term home-cage-like assessments. Furthermore, there was a significant decrease in exploratory behaviors and long-term home-cage-like activities, which were linked to hangover symptoms. This study provides a preliminary approach for assessing hangover behaviors in mice using automated behavioral monitoring, ensuring improved animal welfare, optimised timing, and extended assessment durations. Hence, we propose automated home-cage-like behavioral assessment as an exploratory model for evaluating hangover behaviors in mice, which may serve as a useful tool for future research on the therapeutic efficacy of anti-hangover compounds.

Folk medicine in Thailand has long made use of Pterocarpus indicus Willd. for treating inflammation-related disorders. However, scientific exploration of isolated compounds from P. indicus for improving inflammation-associated sickness conditions and their impact on central nervous system (CNS) safety remain unexplored. The present study initially screened the anti-neuroinflammatory effects of angolensin, a compound isolated from P. indicus heartwood in vitro. Following substantial findings, the efficacy of angolensin was further evaluated in a mouse model of lipopolysaccharide (LPS)-induced sickness behaviors, alongside an assessment of its CNS safety profiles. The anti-neuroinflammatory effects of angolensin were evaluated in LPS-induced BV-2 microglial cells. The effects of angolensin on sickness behaviors were examined in LPS-induced mice using the Laboratory Animal Behaviors Observation, Registration and Analysis System (LABORAS). Proinflammatory cytokine expression in plasma samples of mice was also determined. LABORAS and rotarod tests were conducted to investigate its impact on the CNS. In vitro assessment of the anti-inflammatory activity of angolensin on BV-2 microglial cells revealed a concentration-dependent reduction in the release of LPS-induced nitric oxide (NO) and proinflammatory cytokines (TNF-α and IL-6). At a concentration of 20 µM, angolensin showed comparable results to the positive control, 20 µM minocycline. In mice, angolensin significantly improved LPS-induced sickness behaviors, as indicated by improved home-cage behaviors. Consistent with the in vitro findings, angolensin attenuated the release of proinflammatory cytokines in the plasma of LPS-induced mice. Importantly, angolensin did not induce any adverse effects on locomotion, motor coordination, or general well-being, indicating a favorable CNS safety profile. Overall, these results highlight the anti-inflammatory potential of angolensin in mitigating sickness behaviors in mice, while demonstrating its CNS safety.

Glioblastoma, a fatal brain cancer with limited treatments and poor prognosis, could benefit from targeting the L-type amino acid transporter I (LAT1). LAT1 is essential for cancer cells to acquire necessary amino acids. Tetrahydrocurcumin (THC), a key curcumin derivative, shows potential for glioblastoma treatment. However, its effectiveness is hindered by poor physicochemical and pharmacokinetic properties. Therefore, this study aims to improve the therapeutic efficacy of THC against glioblastoma by chemically modifying it to target LAT1. A novel series of THC-amino acid conjugates were synthesized by conjugating five amino acids: glycine, leucine, isoleucine, and phenylalanine to THC via carbamate bonds. The therapeutic efficacy of THC-amino acid conjugates was further examined in C6 glioma cells, including the role of LAT1 in their therapeutic effects. Among the conjugates tested, THC conjugated with two phenylalanines (THC-di-Phe) showed remarkably higher cytotoxicity against C6 glioma cells (35.8 μM) compared to THC alone (110.7 μM). THC-di-Phe induced cellular death via necrosis and apoptosis, outperforming THC. Additionally, THC-di-Phe inhibited C6 cell proliferation and migration more effectively than THC. Co-incubation of THC-di-Phe with the LAT1 inhibitor 2-Aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH) further increased cellular death. THC-di-Phe also significantly inhibited the P70SK/S6 pathway, regulated by LAT1 inhibitors, more effectively than THC and displayed a similar binding mode with both JX-075 and BCH to the active site of LAT1. Findings suggest the potential role of THC-di-Phe as a LAT1 inhibitor and provide novel insight into its use as a potent antitumor agent in glioma with increased therapeutic efficacy.

The roots of Cymbidium ensifolium yielded a total of 17 compounds, comprising two new compounds (1–2), one new natural product (3), and 14 known compounds (4–17).

Alkaloid analgesics have been associated with adverse effects on the central nervous system (CNS). Therefore, it is crucial to characterize the effects of alkaloid analgesics. Plants rich in lycorine, an alkaloid, have shown promise as analgesics. However, the exploration of their CNS side effects, and analgesic effectiveness remains incomplete. The aim of the present study was to investigate the CNS safety profiles of lycorine and its potential analgesic efficacy. Lycorine (3, 10, and 30 mg/kg, intraperitoneal) did not affect motor coordination, and doses of 3 and 10 mg/kg of lycorine did not lead to any impairment in spontaneous locomotor activity. However, the highest dose (30 mg/kg) demonstrated a significant impairment in rearing behavior and an increase in immobility. The safety doses were subsequently used to assess the analgesic efficacy of lycorine in a mouse model of inflammatory pain. Lycorine (1, 3, and 10 mg/kg, intraperitoneal) demonstrated a dose-dependent reduction in pain-like behaviors in formalin-induced mice. In the in vitro study, lycorine regulated immune cells, suggesting its involvement as a cellular mechanism underlying the suppression of pain-like behaviors observed in the formalin model. Overall, our findings delineate the CNS safety range of lycorine in mice and suggest its potential use as an analgesic.