Dry eye disease (DED) can lead to severe eye discomfort, which manifests as dryness, pain and decreased vision, and significantly affects daily life. This study elucidated the effects of electroacupuncture (EA) intervention on DED and clarified its potential mechanisms through changes in lacrimal gland lipid metabolism and inflammation. A DED mouse model was established by repeated subcutaneous injections of scopolamine hydrobromide, followed by EA intervention. The therapeutic effects of EA intervention on DED were determined based on tear secretion volume (n = 10), corneal fluorescein staining scores (n = 10), corneal stromal (CS) damage (n = 6), and changes in corneal and lacrimal gland morphology (n = 10). Mechanistic endpoints included lacrimal gland lipidomic profiling by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) (n = 6) and assessment of IL-1β and IL-18 expression by immunohistochemistry (n = 6), western blotting (n = 3) and qRT-PCR (n = 6). EA increased tear secretion, reduced corneal fluorescein staining and ameliorated CS and lacrimal gland pathological alterations. Lipidomic analysis identified 81 metabolites, and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis highlighted arachidonic acid metabolism and inflammation-related pathways. EA reduced arachidonic acid and several downstream pro-inflammatory metabolites while partially restoring omega-3 polyunsaturated fatty acids (PUFAs) and their derivatives. EA also reduced lacrimal gland IL-1β and IL-18 expression at the protein and mRNA levels. These findings suggest that EA may ameliorate scopolamine-induced DED, at least in part, through modulation of lacrimal gland lipid homeostasis and suppression of inflammatory responses.

Cognitive impairment and memory decline represent major health concerns affecting older adults worldwide, and systemic metabolic dysfunction plays a critical role in their progression. Black lychee is produced from Litchi chinensis Sonn. through postharvest thermal processing. It contains polyphenolic compounds known for their neuroprotective properties. This study investigated the cognitive-enhancing effects of black lychee in a rat model of scopolamine-induced cognitive impairment. Male rats (n = 8 per group) received scopolamine (2 mg/kg, i.p.) alone or in combination with black lychee at 100, 200, or 400 mg/kg/day for 20 days. Untargeted blood metabolites and the phytochemical composition of black lychee were identified using [1]H-NMR spectroscopy. Cognitive performance was assessed using the passive avoidance test. Phytochemical analysis confirmed that black lychee is rich in polyphenols. All black lychee-treated groups showed a significant increase in step-through latency compared with the scopolamine group. Blood metabolomic analysis revealed alterations in 26 metabolic pathways associated with oxidative stress and energy metabolism. These findings suggest that polyphenols in black lychee attenuate scopolamine-induced cognitive impairment by reducing oxidative stress and restoring key metabolites involved in energy metabolism. These results support the potential of black lychee as a functional food for mitigating age-related cognitive decline.

In Alzheimer's disease (AD), amyloid beta (Aβ) plaques and hyperphosphorylated tau tangles drive neurodegeneration and cognitive decline. Disrupted calcium homeostasis-particularly via transient receptor potential canonical-1 (TRPC1) channels-contributes to tauopathy and disease progression. This study investigated the therapeutic potential of 2-aminoethoxydiphenyl borate (2-APB), a modulator of TRP channels, in a scopolamine-induced rat model of AD. Adult Wistar rats were assigned to six groups: normal control, disease control (scopolamine 2.5 mg/kg), three 2-APB treatment groups (2.5, 5.0 and 10.0 mg/kg) and a donepezil group (5.0 mg/kg), each with six animals. Treatments lasted three weeks. Cognitive performance was assessed using the Morris water maze, memory consolidation and open field tests. Biochemical assays measured acetylcholinesterase (AChE), calcineurin, oxidative stress markers (GSH, MDA) and inflammatory cytokines (TNF-α, IL-6, IL-1β). Protein and gene expression analyses (Western blot, qRT-PCR) evaluated tau, GSK-3β, TRPC1, MARK2, and calcineurin A/PPP3CA, alongside histopathological and immunohistochemical studies. 2-APB treatment significantly reduced pro-inflammatory cytokines, alleviated neuroinflammation, and decreased oxidative stress. Cognitive function improved, correlating with normalized AChE activity and preserved neuronal structure. At the molecular level, 2-APB reduced tau hyperphosphorylation at Ser396 and Thr231, likely through suppression of TRPC1-associated calcium entry and consequent modulation of calcium-dependent kinases (MARK2) and phosphatases (calcineurin), as well as the GSK-3β pathway. However, given that 2-APB is a broad-spectrum calcium signaling modulator - known to act on IP3 receptors, SOCE/ORAI pathways, and multiple TRPC subtypes in addition to TRPC1-the present findings provide supportive evidence linking TRPC1 modulation to the observed neuroprotective effects. Although, the involvement of additional interconnected signaling mechanisms cannot be excluded and these findings should be interpreted within the context of 2-APB's pleiotropic pharmacological profile. Downregulation of TRPC1 and calcineurin A/PPP3CA gene expression supported 2-APB's role in restoring calcium balance and mitigating tau pathology. These results highlight 2-APB's potential in addressing key AD features-tauopathy, oxidative stress, neuroinflammation and cognitive impairment-warranting further preclinical and clinical research as a potential therapeutic for AD and related tauopathies.

Neurodegenerative diseases are characterized by abnormal protein aggregation and oxidative stress, which contribute to neuronal damage and cognitive decline. Aconogonon alpinum, known as Shinga in Korea, is a perennial Polygonaceae plant traditionally used in folk medicine for its anti-inflammatory properties, yet its neurobiological potential remains poorly characterized. In this study, we investigated the amyloid-related processes and cognitive-enhancing effects of A. alpinum extract through bioactivity-guided isolation and pharmacological evaluation. In APPswe-transfected HeLa cells, A. alpinum extract significantly reduced amyloid-β (Aβ)42 secretion in a concentration-dependent manner. In addition, the extract attenuated glutamate-induced oxidative stress in HT-22 hippocampal cells, accompanied by upregulation of brain-derived neurotrophic factor (BDNF) and cAMP response element-binding protein (CREB), indicating enhanced neuronal resilience. In a scopolamine-induced cognitive impairment mouse model, oral administration of A. alpinum extract improved memory performance across behavioral paradigms, including passive avoidance, novel object recognition, and Y-maze tests. These effects were accompanied by modulation of cholinergic signaling, synaptic integrity, neuroplasticity, and amyloidogenic processing in the hippocampus and cortex. Bioactivity-guided isolation identified two flavonol-derived compounds that suppressed β-secretase activity, reduced Aβ40/42 secretion, and modulated amyloid-related processes in vitro. Collectively, these findings suggest that A. alpinum may modulate multiple pathways associated with cognitive dysfunction and may contribute to its cognitive supportive effects.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline, cholinergic dysfunction, and neuroinflammation. Multitarget-directed ligands (MTDLs) are increasingly recognized as promising therapeutic approaches for such multifactorial pathological disorders. Isoxazole-5-one derivatives represent a novel scaffold with potential neuroprotective activity. In current study, the cognitive and anti-inflammatory properties of a newly synthesized isoxazole-5-one derivative, (E)-3-((benzylamino)-methyl)-4-(3,4,5-trimethoxybenzylidene) isoxazol-5-(4H)-one (M5), were evaluated using scopolamine-induced mice models of AD and biochemical analysis, respectively. Behavioral performance was assessed using the Morris Water Maze (MWM) and Y-maze paradigms. Scopolamine administration significantly impaired spatial learning, memory consolidation, and spontaneous alternation behavior, while treatment with M5 produced dose-dependent memory improvements. At the highest dose (10 mg/kg), M5 markedly reduced escape latency and enhanced target quadrant occupancy in the MWM, as well as restored alternation rates in the Y-maze (p < 0.001 vs scopolamine), with effects comparable to donepezil, a standard cholinesterase inhibitor. Biochemical analyses further demonstrated that M5 significantly reduced hippocampal levels of TNF-α and IL-6, while downregulating COX-2 and NF-κB expression, confirming its potent anti-inflammatory action. Taken together, these findings suggest that M5 ameliorates memory impairment potentially through its anti-inflammatory actions. Therefore, M5 may represent might be a promising multitarget-directed ligand for Alzheimer's disease.

Background: Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by oxidative stress, cholinergic dysfunction, and amyloid-β (Aβ) aggregation. Tri-Sannibat-Phol (TSB), a classical Thai polyherbal formulation comprising Piper retrofractum fruit, Ocimum tenuiflorum root, and Piper nigrum root, has been traditionally used for its medicinal properties, yet its anti-AD potential has never been scientifically evaluated. Methods: The therapeutic potential of TSB was investigated through in vitro bioassays including antioxidant, acetylcholinesterase (AChE) inhibitory, and anti-Aβ aggregation assays, alongside neuroprotective evaluation in H2O2-induced SH-SY5Y neuroblastoma cells. Acute oral toxicity was assessed in male ICR mice in accordance with OECD Guideline 420. Cognitive-enhancing effects were evaluated using the modified Y-maze, Novel Object Recognition, and Morris Water Maze tests in a scopolamine-induced amnesic mouse model. LC-MS/MS analysis was performed for phytochemical characterization and chemical standardization of the formulation. Results: TSB demonstrated significant antioxidant activity, AChE inhibitory activity, and anti-Aβ aggregation effects, with P. nigrum and O. tenuiflorum identified as the primary contributing components. Neuroprotective effects were confirmed in H2O2-induced SH-SY5Y cells, where TSB significantly improved cell viability across concentrations of 1-100 µg/mL. Acute oral toxicity assessment revealed an LD50 exceeding 2000 mg/kg, indicating a favorable safety profile. In vivo behavioral studies demonstrated that TSB at medium-to-high doses significantly reversed scopolamine-induced cognitive deficits across all three behavioral tests. LC-MS/MS analysis identified thirteen piperidine alkaloids, with piperine as the dominant constituent at 17.61 ± 0.80% w/w, proposed as the primary bioactive driver and chemical marker for future quality standardization. Conclusions: These findings suggest that TSB exerts multi-targeted anti-AD effects through complementary mechanisms, supporting its potential as a traditional medicine-based therapeutic candidate for further preclinical and clinical investigation.

Dry eye disease (DED) is a multifactorial ocular surface disorder characterized by inflammation, oxidative stress, tear film instability, and secretory dysfunction. This study investigated the protective effects of Lute-gen®, a lutein and zeaxanthin-based formulation, in both in vitro and in vivo DED models. Human corneal epithelial (HCE-T) cells were stimulated with TNF-α, while dry eye was induced in female Sprague-Dawley rats using subcutaneous scopolamine (SCP) administration. In HCE-T cells, Lute-gen® showed no cytotoxicity, restored cell viability, reduced intracellular ROS, and was associated with increased expression of antioxidant-related markers (Nrf2, HO-1, SOD, CAT, and GPx), reduced expression of inflammatory mediators (TLR4/MyD88/NF-κB/NLRP3), and increased expression of AQP3 and AQP5. In SCP-induced rats, Lute-gen® significantly improved tear secretion and reduced corneal fluorescein staining. Histopathological analyses revealed restoration of conjunctival goblet cells, mucin staining, corneal epithelial integrity, acinar area and cell density, and lacrimal gland architecture, with reduced inflammatory infiltration. Immunofluorescence further demonstrated reduced TLR4 and MMP9 immunoreactivity and decreased CD68+ inflammatory cell infiltration. Molecular analyses showed reduced expression of inflammatory cytokines and NF-κB/MAPK/MMP signaling-related inflammatory mediators, together with restoration of AQP1, AQP3, and AQP5 expression in corneal tissues. Collectively, these findings suggest that Lute-gen® treatment was associated with improvements in dry eye-related pathological changes, including restoration of antioxidant-related markers, attenuation of inflammatory responses, restoration of aquaporin expression, and preservation of ocular surface and lacrimal gland integrity. These preclinical findings support further mechanistic investigations and the future clinical evaluation of Lute-gen® as a potential nutritional intervention for dry eye disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder lacking effective disease-modifying therapies. A promising regenerative approach involves enhancing endogenous neurogenic capacity within the injured brain. Reactive astrocytes-stellate-like cells in the AD brain-may contribute to a pro-neurogenic environment through transcription factors (TFs) such as neurogenin 2 (NGN2) and SOX-11. This process is tightly regulated by epigenetic mechanisms, particularly SIRT-1, a neuroprotective histone deacetylase that modulates TF activity and neuronal fate. Vitamin A (VA), a key regulator of differentiation and epigenetic remodeling via its active metabolite retinoic acid, is stored in astrocytes and hepatic stellate cells (HSCs). We hypothesized that AD-related astrocyte activation depletes cerebral VA, mobilizes hepatic stores, contributes to liver fibrosis, and that VA supplementation may restore astrocytic function, activate endogenous TFs via SIRT-1, and drive cholinergic neuron regeneration. In a scopolamine (SCO)-induced AD rat model, VA biodistribution was traced using confocal microscopy. Brain and liver VA deficiency were confirmed via retinol-binding protein (RBP) and ALDH1A1 expressions. Rats received VA (1500, 3000, or 4500 IU/kg/day) or donepezil. Outcomes included neurogenesis (DCX), NGN2/SOX-11 expression, SIRT-1 activation, cholinergic regeneration, amyloid-β deposition, and serum tau. Liver fibrosis was assessed via TGF-β, hydroxyproline and histopathologically. AD induced systemic VA depletion and liver fibrosis. Medium-dose VA (VAMD) significantly enhanced neurogenesis, TF expression, SIRT-1 activation, cholinergic regeneration, and reversed liver fibrosis. VAMD demonstrated neuroregenerative and antifibrotic effects, indicating a possible therapeutic role in AD.

Anisodus tanguticus (Maxim.) Pascher has been documented in Tibetan ethnomedicine. Traditional records indicate that A. tanguticus has traditionally been used to relieve pain, treat parasitic infections, and heal skin wounds caused by viral infections. A. tanguticus is rich in tropane alkaloids, including anisodamine, scopolamine, and atropine, which are pharmacologically active constituents with well-known parasympatholytic effects. These alkaloids have shown promise in controlling cancer cell proliferation and metastasis, with preclinical studies indicating antitumor activity against liver, breast, and colorectal cancers. The extraction of A. tanguticus traditionally involves methods such as juice pressing or boiling; however, modern techniques like ultrasonic, reflux, and supercritical fluid extraction have enhanced alkaloid yield and quality. While the pharmacological properties of A. tanguticus suggest that some constituents of A. tanguticus may have preclinical antitumor relevance, most studies remain preclinical, and clinical data is limited. This review highlights the need for further pharmacological, toxicological, and translational studies. Current evidence suggests that A. tanguticus and several of its constituents exhibit preclinical antitumor activity; however, their clinical relevance remains uncertain, and further pharmacological, toxicological, and clinical validation is required. This narrative review summarizes the traditional use, phytochemical composition, extraction methods, antitumor-related findings, and proposed mechanisms of A. tanguticus, while highlighting the need for further pharmacological, toxicological, and translational studies.

Drug-facilitated sexual assault is a major public health and forensic concern, frequently involving central nervous system depressants such as scopolamine. This tropane alkaloid is especially hazardous due to its strong amnestic, sedative, and hallucinogenic effects, combined with its lack of color, odor, and taste. Despite its toxicological importance, there are still few methods available for the rapid on-site detection of scopolamine. Therefore, portable, low-cost analytical tools capable of detecting scopolamine in a variety of matrices at the point-of-need are still lacking. Here, we developed an autonomous microfluidic paper-based analytical device for the rapid detection of scopolamine in pharmaceuticals, plant extracts, and beverages. The device is based on Scott's colorimetric reaction, in which Cobalt(II) thiocyanate forms a visible pink-to-blue complex with scopolamine under acidic conditions. The system consists of a cellulose substrate sandwiched between hydrophilic and hydrophobic pressure-sensitive adhesive layers and a poly(methyl methacrylate) cover, enabling capillary-driven fluid transport and direct visual readout. Calibration studies demonstrated a concentration-dependent color change, achieving limits of detection of 0.13 mg mL[-1] for pure scopolamine and 0.11 mg mL[-1] for commercial formulations such as Buscapina®. The device was tested in a wide range of alcoholic and non-alcoholic beverages and produced a clear response in extracts of Datura stramonium and Atropa belladonna. This microfluidic paper-based device provides a fast, reliable, and easy-to-use approach for the on-site detection of scopolamine and related compounds. Its low cost, portability, and visual readout make it particularly suitable for forensic screening and public health protection. The platform represents a significant advancement toward practical point-of-need detection of drug-facilitated assault agents.

Alzheimer's disease (AD) is associated with renin angiotensin system (RAS) overactivation that stimulates amyloid β-induced neurodegeneration. Activation of angiotensin II type 1 (AT1) receptor impairs the protective phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway. The combination between angiotensin converting enzyme 2 (ACE2)/ angiotensin 1-7 (Ang 1-7) /Mas receptor (MAS1) axis activation and PI3K/Akt stimulation counteracts AT1-induced apoptosis. Dapagliflozin (DAPA) alters the imbalance between the two RAS arms favoring the protective arm. Accordingly, the aim of the current study was to evaluate the possible neuroprotective effects of DAPA in guarding against scopolamine (SCO)-induced AD in rats via inhibiting RAS classical arm, and activating RAS protective arm, as well as PI3K/Akt pathway. DAPA administration suppressed ACE expression and reduced the levels of Ang II and AT1 in the hippocampi, while elevated the hippocampal levels of ACE2 and Ang 1-7 in addition to boosting MAS1 hippocampal expression. Of note, DAPA prominently augmented the level of PI3K. Also it enhanced the activity of protein kinase A. Furthermore, the hippocampal levels of caspase-3, cytochrome-c, interleukin-6 and nuclear factor kappa B were suppressed, while guanine nucleotide-binding protein alpha-3 level was elevated by DAPA administration. DAPA also attenuated the histopathological SCO-induced damage in rats and enhanced their recognition memory and learning in novel object recognition test and Y-maze spontaneous alternation test. Hence, the current results represent the first evidence for proving that DAPA-induced activation of the protective RAS arm suppressed the cognitive deficits in SCO-induced AD in rats.