ABSTRACT Dysregulated tricarboxylic acid (TCA) cycle activity is increasingly recognized as a contributor to Alzheimer's disease (AD) pathogenesis, yet the mechanistic underpinnings of the relationship remain unclear. Here, we identify isocitrate dehydrogenase 1 (IDH1), a key enzyme in the TCA cycle, as a critical pathogenic driver of AD in microglia. IDH1 expression was markedly upregulated in microglia from both AD patients and 5×FAD mice. Elevated IDH1 promoted excessive cytosolic citrate consumption, which restricted citrate shuttling into mitochondria and impaired mitochondrial TCA cycle function. This citrate metabolic imbalance further disrupted epigenetic regulation, thereby exacerbating AD‐related pathological processes. Using structure‐based screening and co‐crystallization analysis, we identified Kinsenoside (KIN), a natural small molecule, as a selective competitive inhibitor of IDH1 that binds to its isocitrate‐binding pocket. Targeting IDH1 with KIN inhibited its activity, which restored intracellular citrate distribution, reactivated mitochondrial TCA cycle flux, and reestablished metabolic homeostasis. Notably, this intervention not only attenuated neuroinflammation but also reduced β‐amyloid (Aβ) deposition and significantly improved cognitive performance in 5×FAD mice. Collectively, our findings establish IDH1‐mediated metabolic dysregulation as a pivotal pathogenic mechanism in AD and highlight KIN as a promising therapeutic candidate by targeting microglial IDH1 to restore metabolic and functional homeostasis.

BackgroundAlzheimer's disease (AD) is characterized by amyloid-β (Aβ) accumulation with impaired lymphatic clearance, yet therapies targeting lymphatic dysfunction remain underexplored. The Neurolymphatic Formula (NLF), a traditional Chinese medicine, demonstrates lymphatic modulation potential, but its mechanistic role in AD remains unknown.ObjectiveThis study aimed to elucidate NLF's therapeutic efficacy in AD and investigate whether it enhances central and peripheral lymphangiogenesis via VEGF receptor 3 (VEGFR3) activation.MethodsWe evaluated NLF's pharmacological effects on behavior and AD pathology in APP/PS1 mice, including sunitinib-induced lymphatic impairment models. Underlying mechanisms were explored using network pharmacology, molecular docking, and in vitro assays on human lymphatic endothelial cells (HLECs).ResultsIn APP/PS1 mice, 4-week NLF treatment reduced Aβ plaque burden by 43% (p < 0.01) and improved spatial memory latency by 35% (p < 0.05). NLF restored meningeal and mesenteric lymphatic density in sunitinib-treated mice to 82% and 133% of baseline, respectively (p < 0.01), while upregulating serum VEGFR3 2.3-fold (p < 0.01). To validate NLF's molecular basis, coptisine was identified as a representative VEGFR3 ligand (-7.1 Kcal/mol). In vitro, coptisine (25 μM) enhanced HLEC viability by 60%, accelerated wound closure 2.5-fold, and increased tube junctions by 75% (all p < 0.01) alongside VEGFR3 upregulation.ConclusionsNLF alleviates AD pathology by promoting Aβ clearance through VEGFR3-mediated dual modulation of central and peripheral lymphatic systems. The in vitro efficacy of its constituent, coptisine, mechanistically validates this pro-lymphangiogenic pathway, highlighting NLF's therapeutic potential as a holistic lymphatic-targeted AD treatment.

Abstract Objective: Combing, a common daily practice, may promote brain health by improving both extracranial and intracranial blood circulation, as demonstrated in stroke models. However, the optimal combing direction for maximizing circulatory benefits remains unclear. This study was to investigate how the combing direction affects extracranial circulation in healthy adults. Materials and Methods: In this open-label randomized controlled trial, a total of 62 healthy participants were assigned to two groups: Group F–B (front-to-back combing) and Group B–F (back-to-front combing). Participants combed their hair for 5 min using their dominant hand at a consistent frequency, ensuring uniform coverage of the scalp. Blood flow and skin temperature in the ear concha and auricle were measured before and after combing using laser speckle flow imaging and thermometry. Results: Both groups showed significant increases in scalp, ear, and hand skin temperatures. Group B–F exhibited a significant increase in ear blood flow ( P = 0.0027), whereas Group F–B showed no significant change ( P > 0.05). Conclusions: Back-to-front combing significantly enhanced extracranial blood flow, particularly in the ear, indicating that combing direction influenced circulatory outcomes. These findings support back-to-front combing as a simple approach to promoting head circulation within traditional Chinese medicine practices. Clinical trial registration: The study was approved by the Ethics Committee of Beijing University of Chinese Medicine (2020BZHYLL0108), and all participants provided written informed consent.

ObjectiveCerebrospinal fluid (CSF) plays a crucial role in maintaining the homeostasis of the central nervous system (CNS). CSF rapidly exchanges with interstitial fluid (ISF) via the glymphatic system within the brain parenchyma. CSF-ISF circulation and its associated mechanisms are often referred to as the brain lymphatic system. This system is connected directly to meningeal lymphatic vessels (mLVs), jointly performing the function of clearing metabolic waste from the CNS. Emerging evidence indicates that this system is closely associated with the onset and progression of neurodegenerative diseases (NDs) such as Alzheimer’s disease (AD). Importantly, abnormal CSF circulation is not only a downstream consequence of AD pathology, but also a risk factor. In AD, the dynamics of CSF flow within the CNS are diminished, immune dysregulation occurs, and this may increase the risk of AD by exacerbating the burden of amyloid β-protein (Aβ). In the mouse model of AD, impaired CSF flow compromises this clearance function, leading to cognitive deficits. Clinically, acupuncture at cognition-related acupoints is commonly used for the prevention and treatment of AD. However, whether its therapeutic effects are mediated through the modulation of CSF dynamics remains unclear. This study aimed to evaluate the impact of acupuncture on CSF flow and investigate its acupoint specificity.MethodsMice were randomly assigned to experimental groups for the different electroacupuncture groups with the following acupoints: Baihui point (GV 20), Ear point, Neiguan point (PC 6), and Tianshu point (ST 25). Wild-type mice on a C57BL/6J background were used as controls. Fluorescent tracer was injected into the cisterna magna to label CSF flow. Fluorescence imaging was employed to assess the distribution of CSF within the brain before and after acupuncture stimulation.ResultsFollowing tracer injection into the cisterna magna, fluorescence signals rapidly reached the cerebellum and medulla—the regions closest to the injection site. Fluorescence intensity was higher in ventral brain regions compared to dorsal regions, likely due to greater vascular density in ventral areas facilitating CSF-ISF exchange. Electroacupuncture at the GV 20 produced the most pronounced enhancement of CSF across the whole brain, while stimulation at the ST 25 primarily augmented flow within subcortical regions. In contrast, electroacupuncture at the Ear point or the PC 6 had no observable effect on CSF in mice.ConclusionElectroacupuncture promotes CSF flow into the brain parenchyma in an acupoint-specific manner, with GV 20 exhibiting the most pronounced enhancement of CSF dynamics. These findings suggest that acupuncture-mediated facilitation of CSF flow may represent a potential therapeutic strategy for preventing or delaying age-related cognitive decline.

Metabolic dysfunction-associated steatotic liver disease (MASLD) has emerged as a global epidemic, with growing evidence suggesting its adverse impact on brain function. However, the underlying mechanisms linking hepatic metabolic dysfunction to neurodegeneration remain unclear. In this study, we systematically investigated the liver-brain axis by integrating genetic epidemiology, experimental neuroscience, and transcriptomics techniques. Two-sample Mendelian randomization (MR) analysis revealed a potential causal relationship between MASLD and cognitive decline. These findings were validated in a high-fat diet (HFD)-induced MASLD mouse model, which exhibited hallmark features of metabolic dysfunction, including significant body fat accumulation and elevated serum levels of pro-inflammatory cytokines (interleukin-6 [IL-6] and tumor necrosis factor-α[TNF-α]). Behavioral assays demonstrated pronounced anxiety-like behaviors and impaired spatial memory. Neuropathological analysis revealed neuronal loss and structural alterations in the hippocampal dentate gyrus (DG), accompanied by astrocyte remodeling and M1 microglial polarization, indicating neuroinflammation-driven disruption of hippocampal circuits. At the molecular level, MASLD altered the expression of key hippocampal genes-including TCF7L2, LCN2, and AQP1-impacting immune response, lipid metabolism, and apoptotic pathways, which collectively contributed to cognitive deficits. Dual immunofluorescence staining, combined with Sholl and 3D analysis quantitatively characterized neuroglial morphological and functional changes, providing structural-level evidence for MASLD-related brain dysfunction. Taken together, our findings identify MASLD as a modifiable risk factor for neurodegeneration, with systemic inflammation playing a pivotal role in the liver-brain axis. This study highlights key genes and pathways underlying MASLD-induced cognitive impairment, advances understanding of metabolic-neural cross talk, and offers potential therapeutic targets for mitigating cognitive decline through intervention in the liver-brain axis, developing intervention strategies and highlight the therapeutic promise of targeting the liver-brain axis.

BackgroundGlioblastoma (GBM) remains a therapeutic challenge with limited treatment options and poor prognosis. 18β-Glycyrrhetinic acid (GA), a natural metabolite from licorice, has shown anti-tumor potential, but its pharmacological effects in GBM and the underlying mechanisms require systematic investigation.MethodsThe anti-GBM activity of GA in vitro was evaluated in GBM cells using CCK-8, colony formation, wound healing, and flow cytometry assays. Intracranial and subcutaneous GBM models in C57BL/6 and nude mice were established to assess the in vivo pharmacological effects of GA. An integrative approach combining network analysis, transcriptomic sequencing, and TCGA data analysis was employed to explore key genes and pathways. Molecular docking predicted GA binding to MAPK11, and Western blotting assessed its impact on p38 signaling pathway.ResultsGA significantly inhibited GBM cell proliferation, migration, and induced apoptosis in vitro. In vivo, GA treatment markedly suppressed tumor growth in both intracranial and subcutaneous models, with no observed toxicity. Integrated bioinformatics analysis revealed that high MAPK11 (p38-pathway) expression was significantly associated with poor patient prognosis in TCGA. Molecular docking confirmed a strong binding affinity between GA and MAPK11. Mechanistically, GA downregulated MAPK11 expression, activated p38 signaling pathway, and subsequently suppressed the MEK/ERK signaling pathway.ConclusionThis study demonstrats that GA exerted potent anti-GBM effects by regulating p38 signaling pathway, provides novel mechanistic insights, and positions its as a promising therapeutic candidate against GBM.

BACKGROUND:Gefitinib is associated with various adverse reactions, with diarrhea being prevalent. It is mainly managed through lifestyle changes and symptomatic pharmacological interventions, but these approaches have limited effectiveness and frequent recurrence. Qi Yin San Liang San Decoction (QYSLS) shows promise in relieving gefitinib-induced diarrhea, but its mechanisms are unclear. OBJECTIVE:This study aims to explore the pathological mechanisms underlying gefitinib-induced diarrhea and to elucidate the molecular pathways through which QYSLS mediates its therapeutic effects. METHODS:RNA-seq identified differentially expressed genes (DEGs) in colon samples from control and gefitinib-induced diarrhea rats. Network pharmacology was employed to predict the bioactive components and potential targets of QYSLS. A protein-protein interaction (PPI) network was utilized to explore the interactions among these targets, while GO, KEGG, and GSEA enrichment analyses were conducted to reveal the signaling pathways associated with these targets. RNA-seq was used to detect DEGs in QYSLS-mediated relieving of gefitinib-induced diarrhea; the intersection with potential targets was further analyzed to identify key genes. The expression of hub genes was validated through immunohistochemistry and RT-qPCR. RESULTS:RNA-seq and network pharmacology identified 103 bioactive components of QYSLS, with 84 potential targets in QYSLS relieving gefitinib-induced diarrhea. The DEGs in QYSLS relieving gefitinib-induced diarrhea and 84 potential targets were intersected, resulting in the identification of 26 key genes. Further analysis highlighted three central hub genes (CCL20, CCL25, NOS2), which were enriched in pathways related to innate immune response. Furthermore, immunohistochemistry and RT-qPCR confirmed that the expression of CCL25 was reduced by QYSLS in gefitinib-induced diarrhea rats. CONCLUSION:These results indicate that QYSLS may exert its therapeutic effect on gefitinibinduced diarrhea via the modulation of chemokines and innate immune responses.

The emergence of nanotechnology has significantly promoted the development of the medical field, and the development of nanomedicines has broadened the application potential of existing drugs. However, at present, many new types of nanomedicines suffer from poor stability, uncertain safety, complex preparation processes, and high costs. The development of nano- Chinese herbal medicines (CHMs) has provided new ideas for the application of nanomedicines. Nano-CHMs are simple to prepare, easily accessible, and possess stable and reliable safety. This review highlighted nanoparticles with pharmacological activities are the important material basis for CHM formula decoctions. Given the complex composition of CHM formulas, the article further presented the nanoaggregates and exosome-like vesicle nanostructures in single herbs, and self-assembly effect of natural compounds. For the first time, this article systematically introduced the composition and characteristics of nanoparticles from CHM formulas to single herbs and then to natural compounds, from nano-aggregates with unclear components to exosome-like vesicle structures and then to self-assembled structures of supramolecular materials. It also summarized the problems to be solved in the research of CHMs nanoparticles and the potential for further applications, aiming to provide guidance for the modernization of CHMs -related nanoparticles.

Central nervous system (CNS) diseases, including neurodegenerative diseases, stroke, brain tumors, and others, result in poor quality of life and can cause substantial disability. Not all CNS diseases are amenable to surgical approaches, so drug development is important for disease treatment. Unfortunately, there are few drugs currently available for CNS diseases. Chinese herbal medicines (CHMs), represented by herb derived active ingredients and decoctions, have long been used in the treatment of CNS diseases. One of the major challenges in the further development of CHMs for CNS diseases is to improve cerebral drug delivery, and nanotechnology applied to CHMs to obtain nano-Chinese Herbal Medicines (nano-CHMs) is an effective way. This review categorizes nano-CHMs according to their preparation methods and sources, and discusses the applications of each nano-CHM and its advantages. Considering the special status of the brain, we further summarize the delivery strategies of nano-CHMs and the physiological barriers that need to be overcome for different strategies. Finally, we discuss the application of nano-CHMs for various CNS diseases, aiming to provide insights into the development of nano-CHMs for the treatment of CNS diseases.

Objective: Alzheimer’s Disease (AD) is a progressive neurodegenerative disorder with limited options for reversing its middle-to-late stages. Early intervention is crucial to slow down disease progression. This study aimed to investigate the potential of the NeuroProtect (NP) formula, a combination of geniposide and Panax notoginseng saponins, in preventing AD. We evaluated the effects of the NP formula on amyloid plaque accumulation, neuronal degeneration, and molecular signaling pathways using in vivo and in vitro models. Methods: To predict functional pathways and potential downstream targets of NP intervention, we employed network pharmacology. The preventative impact of the NP formula was assessed using APP/PS1 mice. We conducted HE staining, ELISA assay, Golgi staining, and immunohistochemistry to detect the protective effect of NP. Additionally, cell experiments were performed to assess cell activity and target protein expression. Results: Network pharmacology analysis revealed 145 drug-disease interactions and identified 5 core active targets associated with AD. Molecular docking results demonstrated strong binding affinity between the components of the NP formula (GP, GN-Rb1, GN-Rg1, NS-R1) and target proteins (STAT3, HIF1A, TLR4, mTOR, VEGFA). Notably, the binding energy between NS-R1 and mTOR was -11.4kcal/mol. Among the top 10 enriched KEGG pathways, the HIF-1 and PI3K-AKT signaling pathways were highlighted. In vivo experiments demonstrated that the NP formula significantly ameliorated pathological changes, decreased the Aβ42/Aβ40 ratio in the hippocampus and cortex, and increased dendritic spine density in the CA1 region during the early stage of AD. In vitro experiments further illustrated the NP formula’s ability to reverse the inhibitory effects of Aβ25-35 on cell viability and regulate the expression of Tlr4, Mtor, Hif1a, Stat3, and Vegfa. Conclusion: Our findings suggest that NP exhibits neuroprotective effects during the early stages of AD, positioning it as a potential candidate for AD prevention. The NP formula may exert its preventive effects through the HIF-1/PI3K-AKT signaling pathway, with mTOR identified as a key target.

Drug discovery before the 20th century often focused on single genes, molecules, cells, or organs, failing to capture the complexity of biological systems. The emergence of protein-protein interaction network studies in 2001 marked a turning point and promoted a holistic approach that considers the human body as an interconnected system. This is particularly evident in the study of bidirectional interactions between the central nervous system (CNS) and peripheral organs, which are critical for understanding health and disease. Understanding these complex interactions requires integrating multi-scale, heterogeneous data from molecular to organ levels, encompassing both omics (e.g., genomics, proteomics, microbiomics) and non-omics data (e.g., imaging, clinical phenotypes). Artificial intelligence (AI), particularly multi-modal models, has demonstrated significant potential in analyzing CNS-peripheral organ interactions by processing vast, heterogeneous datasets. Specifically, AI facilitates the identification of biomarkers, prediction of therapeutic targets, and simulation of drug effects on multi-organ systems, thereby paving the way for novel therapeutic strategies. This review highlights AI's transformative role in CNS-peripheral interaction research, focusing on its applications in unraveling disease mechanisms, discovering drug targets, and optimizing clinical trials through patient stratification and adaptive trial design.

Over centuries of clinical practice, Chinese herbal medicines (CHMs) have gained widespread recognition for their efficacy in treating various diseases. However, their complex material basis and relatively mild therapeutic efficacy limit their modernization and quality control. Recently, the application of pharmaceutical nanotechnology to CHMs has not only enhanced their efficacy, but also helped elucidate their material basis, thereby substantially advancing their modernization. Nano-modified CHMs have shown improvements in multiple aspects, such as bioavailability, targeting ability, toxicity reduction, and sustained release. In this review, nano-strategies for emerging, revolutionary, and promising pathways for modernizing CHMs are revealed. First, the development, application potential, particularities, and limitations of CHMs are reviewed. Subsequently, a systematic and comprehensive analysis of the two nano-strategies for optimizing CHMs are presented, highlighting the distinct characteristics of the carrier-free and carrier-based approaches. The specific advantages of these strategies, including improved bioavailability, increased targeting ability, reduced toxicity, and controlled release, are discussed. The novel research directions resulting from the application of pharmaceutical nanotechnology to CHMs are also explored, such as elucidating CHM treatment theories, combining traditional Chinese medicine topical therapies, drug screening, and expanding innovative drug formulations. Finally, the challenges and opportunities in this field are addressed to inspire future research.

Background Alzheimer's disease (AD) is recognized as a multifactorial neurodegenerative disorder involving numerous cellular and molecular processes, such as sleep disturbance, imbalance in brain glucose metabolism and neuroinflammation; these dysregulations typically precede the onset of symptoms. Hence, the results of mono-target therapy after AD diagnosis are in many cases unsatisfactory. Objective Traditional Chinese medicine (TCM) presents significant potential for treating AD. Sleep disorders are one of the early symptoms of AD; however, there is no effective solution to sleep disorders caused by AD. Some TCMs have been shown to treat sleep disorders by regulating energy metabolism or improving neuroinflammation. This study aims to investigate if XX-F administrated in advance could alleviate AD by improving sleep quality and neuroinflammation. Methods Mice were given Xiexintongfu formula (XX-F) intragastrically for three months. Morris water maze and pentobarbital-induced sleep test were performed to evaluate cognition and sleep. Determine changes in energy metabolism related to glycolysis through western blot and specific assay kits. Using immunofluorescence and western blot to detect neuroinflammation. Results Shortened sleep duration and cognitive impairment were observed in 6-month-old APP/PS1 mice. XX-F significantly prolonged sleep duration and rescued cognition. In addition, XX-F reduced the number of amyloid-β (Aβ) plaques and ameliorated neuroinflammation, and inhibited glycolysis by reducing pyruvate kinase M2 (PKM2) and lactate levels while rescuing adenosine triphosphate (ATP) deficiency. Conclusions We demonstrate that XX-F can improve sleep and cognition of AD mice by regulating energy metabolism and reducing neuroinflammation. This is a potential treatment method for AD and requires further in-depth research.

This study examined the longitudinal association of metabolic dysfunction-associated fatty liver disease (MAFLD) with distinct cognitive function trajectories, and determine whether and to what extent this association was mediated by MAFLD-related metabolites among 845 participants. Two cognitive function trajectories were identified as normal (n = 714, 84.50%) or large decrease (n = 131, 15.50%) pattern over 7 years. Participants with MAFLD (N = 277, 32.78%) had an 81% higher risk of developing a large decrease in cognitive function (odds ratio, 1.81; 95% confidence interval, 1.16–2.94) than non-MAFLD. Three MAFLD-related metabolites were identified as lysoPC(20:3(5z,8z,11z)), lysoPE(18:1(9z)/0:0), and valine, of which lysoPE(18:1(9z)/0:0) and valine played a partially mediated role in the association of MAFLD with a large decrease in cognitive function (mediation proportion = 9.93% and 11.04%, respectively). The results indicated that MAFLD was associated with a higher risk of developing a large decrease in cognitive function, which was partially mediated by lipid and amino acid metabolism.

Cancer treatment is often ineffective due to poor bioimaging and resistance to standard therapies. This issue is exacerbated by multiple low-penetrable bio-barriers that limit the theranostic agents' effectiveness in tumors. Here, a hollow nanomotor PM-HMSN/Arg is fabricated by a sequential process involving: electrostatic adsorption of Mn2+, loading of l-Arg, and coating of platelet membrane (PM), respectively. This nanomotor uses l-Arg as an NO donor and ultrasound (US) as a trigger for NO release. After administration, it improves tumor penetration via a "tethering-relaxing-drilling" mechanism, overcoming bio-barriers during delivery from blood vessels to tumor cells. NO regulates the metabolism of tumor vascular endothelial cells, facilitating relaxation, and enhances cytotoxicity by participating in reactive oxygen species metabolism. More importantly, the nanomotor's active motion enhances tissue penetration and retention in cancer, increasing therapeutic effects. In addition, continuous in situ NO generation extends US imaging signal lifetime. This innovative nanomotor shows promise for multimodal theranostics in low-penetrable tumors.

AIMS:To investigate the therapeutic effects of kinsenoside on cognitive dysfunction in APP/PS1 transgenic mice and to explore its potential targets. MATERIALS AND METHODS:Network pharmacology was employed to identify targets of kinsenoside in Alzheimer's disease (AD). The Morris water maze test was conducted to assess the therapeutic effects of kinsenoside on cognitive dysfunction in APP/PS1 transgenic mice, and immunofluorescence was used to evaluate the impact of kinsenoside on amyloid-beta (Aβ) pathology and brain lymphatic structure and function. The Raybiotech GSM-CAA-4000 protein chip was used to detect changes in inflammatory factors. Graph convolutional network (GCN) analysis was applied to analyze and identify core targets from the protein chip results, which were then intersected with the network pharmacology findings, followed by molecular docking and molecular dynamics simulations. RESULTS:Network pharmacology analysis indicated that kinsenoside inhibits inflammation and oxidative stress signaling pathways involved in AD. The maximum neighborhood connectivity (MNC) algorithm identified HSP90AA1, HSP90AB1, PIK3CA, STAT3, IL6, and IFNγ as potential targets. In vivo mouse experiments demonstrated that kinsenoside has the ability to reduce Aβ plaque deposition and expand meningeal lymphatic vessels(mLVs), improving the glymphatic system drainage, which ultimately leads to improved cognitive function. This beneficial effect may be related to the inhibition of IFNγ by kinsenoside. CONCLUSION:Based on preclinical data, kinsenoside shows promising potential in the treatment of AD.

To investigate the distribution and characteristics of lymphatic vessels within the central nervous system, we focus on the meninges of the spinal cord and brain parenchyma in mice. Additionally, we aim to provide experimental methods for obtaining optimal imaging and clear structures of lymphatic vessels, while optimizing the perfusion parameters to improve histomorphological quality. Male C57BL/6J mice were randomly divided into four groups, with each group assigned a specific perfusion parameter based on perfusion volumes and temperatures. Immunofluorescence staining of lymphatics and blood vessels was performed on both meningeal and the brain tissue samples. Statistical analysis was performed using one-way analysis of variance to compare the groups, and a significant level of P < 0.05 was considered statistically significant. Our study reports the presence of lymphatic vessels in the meninges of the spinal cord and brain parenchyma in mice. We highlight the crucial role of high perfusion volume of paraformaldehyde with low temperature in fixation for achieving optimal results. We provide experimental methods for obtaining optimal imaging and clear structures of lymphatic vessels in the meninges of the spinal cord and brain parenchyma in mice, which contribute to our understanding of the distribution and characteristics of lymphatic vessels within the central nervous system. Further research is warranted to explore the functional implications of these lymphatic vessels and their potential therapeutic significance in neurodegenerative and neuroinflammatory diseases.

Background: Presenilin (PSEN, PS) is essential for γ-secretase function, and mutations can disrupt amyloid-β (Aβ) production in familial Alzheimer’s disease. Targeting γ-secretase is complex due to its broad involvement in physiological processes. Objective: Our aim was to create a novel knockin (KI) mouse model expressing PSEN1 D385A mutation and investigate the efficacy of a Geniposide and Ginsenoside Rg1 combination (NeuroProtect modified formula, NP-2) in restoring γ-secretase activity. Methods: Using gene manipulation, we established the PS1 D385A KI mouse model and confirmed the mutation, mRNA, and protein levels using Southern blotting, northern blotting, and western blotting, respectively. In vitro γ-secretase assay was conducted to measure γ-secretase activity, while histological analyses examined neurogenesis effects. NP-2 administration evaluated its impact on γ-secretase activity. Results: The PS1 D385A KI homozygotes displayed severe cerebral hemorrhage, postnatal lethality, developmental disorders, reduced proliferation of neural progenitor cells, and disrupted γ-secretase function. The mutation abolished PS1 protein self-shearing, leading to compromised γ-secretase activity. NP-2 intervention effectively restored γ-secretase activity in the heterozygous mice. Conclusions: PS1 D385A mutant disrupted PS1 protein self-cleaving, impairing γ-secretase activity in KI mice. NP-2 restored γ-secretase function, offering potential for novel AD treatment strategies despite the challenges posed by γ-secretase’s complex role in physiological processes.