This study aims to investigate the effects of Naotaifang(NTF) on neuronal ferroptosis and pyroptosis in cerebral ischemia-reperfusion injury(CIRI), with a focus on nuclear factor erythroid 2-related factor 2(Nrf2). A rat model of CIRI was established by middle cerebral artery occlusion/reperfusion(MCAO/R). Sprague-Dawley rats were randomly assigned to a sham group, a model group, low/medium/high-dose(4.5, 9, 18 g·kg~(-1), respectively) NTF groups, and a butylphthalide(60 mg·kg~(-1)) group(n=15 per group). After modeling, the treatment groups received corresponding agents by gavage once daily for 7 days, and the sham and model groups received an equal volume of normal saline. After treatment, the neurological function was assessed based on the Zea-Longa score, the infarct volume by TTC staining, and cellular morphology by HE staining. The levels of Fe~(2+), interleukin-1β(IL-1β), and interleukin-18(IL-18) were measured by ELISA, and those of malondialdehyde(MDA) and glutathione(GSH) by biochemical assays. Western blot was employed to examine the expression levels of Nrf2, glutathione peroxidase 4(GPX4), NOD-like receptor family pyrin domain-containing 3(NLRP3), and the N-terminal fragment of gasdermin D(GSDMD-N). In the cell experiment, oxygen-glucose deprivation/reperfusion(OGD/R) was used to model CIRI in SH-SY5Y cells. siRNA-mediated Nrf2 knockdown(si-Nrf2) was employed to probe into the role of Nrf2 in OGD/R-induced ferroptosis and pyroptosis and the intervention effect of NTF. Membrane damage was evaluated by Hoechst/propidium iodide staining and LDH release, and GPX4 and GSDMD immunofluorescence was employed to indicate ferroptosis and pyroptosis, respectively. Fe~(2+), MDA, and GSH were quantified biochemically, and the levels of IL-1β and IL-18 by ELISA. The expression of Nrf2, GPX4, NLRP3, and GSDMD-N was quantified by Western blot. The cellular ultrastructure was observed by transmission electron microscopy. The results of the animal experiment showed that NTF promoted the expression of Nrf2 to alleviate neuronal ferroptosis and pyroptosis, thus producing significant neuroprotection against CIRI in a dose-dependent manner. The results of the cell experiment showed that the neuroprotective effects of NTF after CIRI were mediated by Nrf2-dependent suppression of neuronal ferroptosis and pyroptosis. Taken together, NTF attenuates CIRI by suppressing neuronal ferroptosis and pyroptosis through Nrf2-mediated regulation.

Background Rheumatoid arthritis (RA)-associated bone erosion is largely driven by excessive osteoclast differentiation and activation. Total saponins of Panax japonicus C.A. Meyer (TSPJs), a ginseng-derived botanical fraction, show anti-inflammatory activity in RA-related settings, but their effects on osteoclast-mediated bone destruction remain unclear. Methods Transcriptomic datasets from RA synovial tissues and RANKL-induced osteoclast differentiation were integrated to identify shared pathogenic signals. TSPJs were chemically profiled by HPLC and evaluated in collagen-induced arthritis mice and RANKL-stimulated RAW264.7 cells. Osteoclast formation and function were assessed by TRAP staining, F-actin ring analysis, gene expression, bone resorption assays, histopathology, radiography, and serological measurements. Network pharmacology, molecular docking, CETSA, immunofluorescence, Western blotting, and immunohistochemistry were used to explore candidate mechanisms. Results Integrated bioinformatics highlighted NF-κB-related signaling as a shared pathway linking RA and osteoclastogenesis. HPLC identified four representative saponins: araloside A, chikusetsusaponin IVa, ginsenoside Rg2, and ginsenoside Ro. In CIA mice, TSPJs reduced arthritis severity, improved joint pathology, lowered osteoclast-associated serum markers, and attenuated radiographic and histological bone erosion. In vitro, TSPJs inhibited RANKL-induced osteoclast differentiation, disrupted F-actin rings, downregulated osteoclast-related genes, and reduced bone resorption. The overall evidence was consistent with modulation of SRC/TRAF6/NF-κB signaling, while docking and CETSA supported SRC/NFATc1/FOS-related nodes as candidates for further validation. Conclusion TSPJs attenuated RA-associated bone erosion and restrained osteoclastogenesis, supporting further evaluation of this Panax japonicus-derived saponin preparation.

Ischemia-reperfusion injury, a critical pathophysiological phenomenon in multiple organ systems, remains a formidable therapeutic challenge in clinical practice. As the third endogenously produced gaseous signaling molecule, hydrogen sulfide (H2S) has emerged as a pivotal regulator of diverse physiological processes and pathological cascades. Accumulating evidence indicates that H2S exerts cytoprotective effects against cerebral, cardiac, hepatic, renal, and pulmonary ischemia-reperfusion injuries through multifaceted mechanisms involving mitigation of inflammatory responses, suppression of oxidative stress, modulation of autophagic processes, and inhibition of apoptotic pathways. This comprehensive review systematically examines the endogenous biosynthesis and metabolic regulation of H2S, while elucidating the molecular mechanisms underlying its organ protective effects during ischemia-reperfusion injury. Particular emphasis is placed on the therapeutic potential of H2S synthase isoforms and bioactive metabolites in ischemic pathophysiology. Notably, recent advances in H2S pharmacology have catalyzed the development of novel H2S donors and slow-releasing compounds, including HSDF-NH2, S-allyl cysteine, S-propargyl cysteine, and S-(4-fluorobenzyl)-N-(3,4,5-trimethoxybenzoyl)-L-cysteine. These pharmacological innovations demonstrate enhanced tissue specificity and controlled release kinetics, paving the way for clinical translation of H2S-based therapeutics in ischemia-reperfusion injury management. Future research directions should focus on optimizing drug delivery systems and elucidating the spatiotemporal dynamics of H2S signaling in organ-specific ischemia-reperfusion pathologies.

The central nervous system (CNS) harbors a distinct immune memory programming system, wherein immunogenic cell death (ICD) acts as a pivotal signaling hub. A spectrum of insults, from systemic metabolic dysfunction to local protein aggregation and ionic dyshomeostasis, can provoke ICD in neurons, glia, and resident immune cells. This process orchestrates the release of damage-associated molecular patterns (DAMPs) from distinct subcellular compartments. These DAMPs synergistically initiate both innate trained immunity (TI), characterized by profound metabolic-epigenetic reprogramming, and antigen-specific adaptive immune responses that traverse the blood-brain barrier. Together, these pathways constitute an integral network of central immune surveillance. Crucially, this ICD-driven immune programming exhibits a striking functional dichotomy depending on the pathological context. In non-neoplastic conditions such as neural injury and neurodegenerative diseases, uncontrolled ICD signaling can establish a pathological trained immune memory, driving a self-perpetuating cycle of chronic neuroinflammation and tissue damage. Conversely, within the tumor microenvironment of malignancies like glioma, the adaptive immune responses elicited by ICD are frequently subverted by potent immunosuppressive mechanisms, culminating in tumor immune escape. This review dissects the differential regulatory mechanisms of ICD-mediated immune memory in CNS tumors versus non-tumor diseases. We aim to elucidate the molecular switches that govern the transition of this immune program from a beneficial, compensatory state to a pathological, detrimental phenotype. By exploring emerging therapeutic strategies, including gene editing, nanomaterials, and bioactive phytochemicals that precisely target ICD pathways, we provide a theoretical framework for understanding CNS immune homeostasis and for the rational design of precision immunotherapies.

With the accelerated pace of global population aging, the number of people suffering from age-related diseases is increasing, posing a serious threat to human health and well-being. Age-related diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), hypertension, atherosclerosis, type 2 diabetes (T2DM), and osteoporosis (OP), are often characterized by physiological function decline and metabolic disorders caused by aging, among which dysregulation of metal ion homeostasis, especially copper homeostasis imbalance, has emerged as a major factor. Copper is an essential enzymatic cofactor whose homeostasis is tightly regulated at systemic, cellular, and subcellular levels. However, during the aging process, the balance between copper uptake and efflux becomes compromised, alongside a reduction in cellular copper-buffering capacity. These alterations may lead to the loss of copper homeostasis and induce cuproptosis, which is a recently elucidated form of regulated cell death (RCD) triggered by copper overload and disrupts mitochondrial metabolism by promoting the aggregation of lipoylated proteins in the tricarboxylic acid cycle (TCA) and destabilizing respiratory chain complexes. Copper homeostasis imbalance and the resulting cuproptosis accelerate the aging process by promoting molecular mechanisms, including telomere attrition, mitochondrial dysfunction, oxidative stress, proteostasis imbalance, epigenetic changes, and chronic inflammation. This review focuses on the reciprocal interactions between aging and copper homeostasis: it elucidates how aging impairs copper homeostatic regulation, and how dysregulated copper metabolism and subsequent cuproptosis accelerate aging and exacerbate age-related diseases. Furthermore, it explores potential therapeutic strategies targeting copper homeostasis and cuproptosis to treat age-related diseases.

Ischemia-hypoxia-induced inflammation and glycolysis are linked to the severity of cerebral ischemia-reperfusion injury (CIRI), but the mechanisms are unclear. Current research suggests that the inflammatory response of immune cells activated by STING is a key regulatory molecule in cellular inflammatory damage. However, the specific mechanisms underlying STING-mediated CIRI inflammatory responses remain unclear. This study found that STING expression was specifically elevated in microglia in the damaged side of the hippocampus in CIRI model mice, and this elevation was positively correlated with the severity of CIRI. Our previous research indicated that the dynamic process of mitochondrial fusion and fission is closely associated with CIRI. Building on this, we integrated glycolysis, mitochondrial fission, and the STING inflammatory pathway. Mechanistically, our data suggest that DRP1 K616 is a critical candidate site involved in DRP1 lactylation-associated regulation, which promotes STING pathway activation and contributes to the progression of CIRI. In conclusion, our findings offer substantial evidence that lactate-driven DRP1-mediated mitochondrial fission facilitates the involvement of the STING inflammatory pathway in CIRI. These results suggest that modulating lactate metabolism may serve as a crucial upstream strategy for therapeutic intervention in CIRI.

Cerebral Small Vessel Disease (CSVD) is a dynamic whole-brain disease, characterized by pathological cascades that affect the brain's venules, capillaries, small arteries, and arterioles. Neuroimaging features of CSVD typically comprise Recent Small Subcortical Infarcts (RSSI), lacunes of presumed vascular origin, White Matter Hyperintensities (WMH) of presumed vascular origin, enlarged Perivascular Spaces (PVS), Cerebral Microbleeds (CMB), and Brain Atrophy (BA). The main clinical features of CSVD often include stroke, abnormal gait, psychiatric disorders, cognitive decline, and urinary incontinence, imposing a heavy burden on individuals and society. Despite its impact, the pathogenesis of CSVD remains unclear, and current clinical diagnosis relies primarily on neuroimaging, presenting considerable challenges for effective treatment. In recent years, most studies have addressed the pathophysiological and molecular mechanisms of CSVD, including chronic cerebral hypoperfusion, inflammatory cascades, oxidative stress, endothelial dysfunction, and Blood-Brain Barrier (BBB) leakage. In addition, genetic factors have been strongly associated with CSVD, though genetic heterogeneity and the complexity of internal environment homeostasis contribute to the persistent uncertainty surrounding its exact mechanisms. A comprehensive overview of these individual mechanisms is crucial for a holistic understanding of the pathogenesis of CSVD. Currently, there is a relative lack of therapeutic drugs and interventions for the complex pathogenesis of CSVD. The existing treatments, such as antihypertensives, antiplatelet agents, lipid-lowering drugs, and hypoglycemic agents, along with traditional alternative therapies like Chinese herbal medicine and acupuncture, have demonstrated efficacy in modulating the occurrence and progression of CSVD. These therapies provide a new perspective for developing more rational CSVD prevention strategies and treatment plans. This review systematically summarizes the cutting-edge research achievements in the field of pathological and physiological mechanisms of CSVD over the past few years, as well as potential treatment pathways and limitations, to provide a theoretical basis and intervention directions for the diagnosis and treatment of this patient population.

Zinc finger proteins (ZFPs), a vast superfamily of sequence-specific DNA and RNA-binding proteins, serve as master regulators of gene expression and cellular homeostasis. While traditionally studied for their roles in development, ZFPs have emerged as critical effectors and therapeutic targets across a wide spectrum of human pathologies, including cancer, neurological disorders, and autoimmune diseases. This review systematically dissects the molecular mechanisms by which dysregulated ZFP activity drives disease pathogenesis, using ischemic stroke as a central exemplar to illustrate their multifaceted roles. We detail how specific ZFPs orchestrate key stroke risk factors such as hypertension, hyperglycemia, and atherosclerosis, subsequently govern post-ischemic injury cascades, including neuroinflammation, programmed cell death, and blood–brain barrier disruption. Addressing the long-standing challenge of ZFPs as “undruggable” targets, we critically evaluate cutting-edge therapeutic strategies poised to modulate their function with precision. These include small-molecule modulators, targeted protein degraders (PROTACs), zinc finger nuclease (ZFN)-based gene editing, and advanced nanocarrier delivery systems, complemented by high-throughput computational screening. By integrating deep mechanistic insights with novel translational approaches, this review establishes a pioneering pan-disease framework for targeting ZFP networks. We provide a structured roadmap for future research and highlight the immense potential of ZFPs as a new class of master regulatory targets for developing novel and feasible therapies in ischemic stroke and beyond.

BACKGROUND:Mitophagy is vital for preventing cerebral ischemia-reperfusion (CI/R) injury. Naringin, a flavanone glycoside, reduces CI/R injury by blocking harmful mitophagy induced by peroxynitrite (ONOO⁻). Our earlier studies identified that mitochondrial calcium (mtCa2+) efflux stabilizes mitochondrial endosymbiosis and initiates protective mitophagy. However, it is unclear whether and how naringin inhibits harmful mitophagy in association with mtCa2+. PURPOSE:This study aimed to examine the influence of the S-nitrosylated dynamin-related protein 1 (SNO-DRP1)/leucine-rich repeat kinase 2 (LRRK2)/mitochondrial calcium uniporter (MCU) pathway on harmful mitophagy, which is inhibited by naringin in CI/R injury. METHODS:To validate the hypotheses, a transient middle cerebral artery occlusion/reperfusion (tMCAO/R) model in rats and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced PC12 cells were used to investigate the effects of naringin. Specifically, linsidomine hydrochloride (SIN-1) was used to enhance SNO-DRP1, whereas spermine (Spm) was used to activate MCU. Mitochondrial fragmentation was assessed by measuring the average mitochondrial diameter via transmission electron microscopy (TEM) and evaluating the protein levels of mitofusin 1 (MFN1) and dynamin-related protein 1 (DRP1) through western blotting (WB). The SNO-DRP1 formation was determined by a biotin switch assay. The extent of SNO-DRP1-mediated excessive mitophagy was examined by quantifying the number of autophagosomes using TEM and assessing the colocalization of translocase of the outer mitochondrial membrane complex subunit 20 (TOMM20)-DRP1-3-NT and 3-NT-DRP1-LC3 via immunofluorescence (IF). The interaction between SNO-DRP1 and LRRK2 was confirmed using co-immunoprecipitation and IF colocalization. MtCa2+ overload induced by MCU was evaluated by analyzing MCU protein levels, measuring mtCa2+ fluorescence intensity following Rhod-2 AM treatment, and assessing the opening of the mitochondrial permeability transition pore. The mitochondrial reactive oxygen species and mitochondrial DNA release were used to evaluate the mitochondrial endosymbiosis failure. RESULTS:Naringin exhibited a dose-dependent protective effect against CI/R injury in rats subjected to the tMCAO/R model. It effectively reduced mitochondrial fragmentation and SNO-DRP1-mediated mitophagy, attenuated the interaction between SNO-DRP1 and LRRK2, and decreased the protein level of MCU in the penumbra region of the cortex. SIN-1 and Spm counteracted the neuronal protection of naringin in OGD/R-induced PC12 cells. They also mitigated the impact of naringin on mitochondrial fragmentation, harmful mitophagy, and MCU-mediated mtCa2+ overload and mitochondrial endosymbiosis disruption. Notably, only SIN-1 reversed naringin's suppression of SNO-DRP1 formation and its interaction with LRRK2 in vitro. CONCLUSIONS:This study demonstrates that naringin reduces ONOO⁻-caused harmful mitophagy in CI/R injury by blocking the SNO-DRP1/LRRK2/MCU pathway, which helps prevent mitochondrial fragmentation and preserve mitochondrial endosymbiosis. This pathway may be among the mechanisms by which naringin exerts its protective effects.

Introduction: This study aimed to investigate whether activating the Nrf2/TFAM pathway boosts mitochondrial biogenesis, reduces ferroptosis in ischemic stroke (IS), and evaluates Naotaifang (NTF) formula's therapeutic potential. Methods: Ferroptosis and mitochondrial biogenesis indicators were measured at various time points following MCAO. Various methods, including transmission electron microscopy, immunofluorescence assay, enzyme-linked immunosorbent assay, Western blotting assays, and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), were employed to evaluate the impact of NTF on mitochondrial biogenesis and ferroptosis in vivo and in vitro. Results: IS significantly inhibits mitochondrial biogenesis and increases neuronal ferroptosis, with brain damage worsening over time. MCAO groups showed reduced expression of Nrf2, TFAM, ATP, CISD2, FPN, GPX4, SOD, and HO-1, alongside elevated Fe²⁺, ROS, and LPO (P < 0.05) compared to the control group. Both sulforaphane and NTF treatment reversed these effects; NTF treatment effectively increased the expression of Nrf2, TFAM, FtMt, CISD1, CISD2, FPN, and GPX4 while inhibiting the levels of Fe2+, ROS, and LPO. (P < 0.05). Discussion: This finding clarifies mitochondrial biogenesis's crucial role, proposes a new "pathway + molecule" strategy for IS treatment, and supports NTF's clinical potential, though larger animal models and long-term safety studies are needed. Conclusion: In the context of IS, reduced mitochondrial biogenesis plays an important role in ferroptosis. Targeting the Nrf2/TFAM signaling pathway may improve mitochondrial biogenesis in IS. Furthermore, NTF can mitigate ferroptosis by promoting mitochondrial biogenesis through the Nrf2/TFAM signaling pathway

Cerebral edema is a life-threatening complication of stroke, yet its underlying mechanisms remain incompletely understood, hindering the development of effective therapies. Aquaporin-4 (AQP4), the principal water channel of the glymphatic system, is crucial for cerebral water homeostasis. Following a stroke, AQP4 undergoes pathological depolarization—a loss of its polarized localization on astrocytic endfeet. This review establishes AQP4 depolarization as a central, unifying event that orchestrates secondary brain injury in stroke. We systematically dissect how AQP4 depolarization disrupts both glymphatic clearance and blood-brain barrier (BBB) integrity, thereby fueling a vicious cycle of edema formation. A key insight we advance is the divergent pathophysiology between stroke subtypes: in ischemic stroke, AQP4 depolarization drives initial glymphatic failure and cytotoxic edema, which subsequently promotes BBB breakdown and vasogenic edema. Conversely, in hemorrhagic stroke, primary vasogenic edema triggers AQP4 depolarization, which then mediates secondary glymphatic impairment and cytotoxic edema. Based on this mechanistic framework, we critically evaluate emerging interventions aimed at preserving AQP4 polarity. We conclude that maintaining AQP4 polarization, rather than non-selectively inhibiting its channel function, represents a paradigm shift and a more precise neuroprotective strategy for combating stroke-induced cerebral edema.

Background Cerebral ischemia-reperfusion injury (CIRI) leads to severe mitochondrial dysfunction, which is a critical trigger of widespread neuronal apoptosis. Therefore, restoring mitochondrial homeostasis represents a key strategy for neuroprotection. Clinical observations suggest that the herbal pair Geum japonicum Thunb. var. chinense-P. decorata H. Andres (GJ-PD) shows therapeutic advantages in alleviating CIRI. However, its precise neuroprotective effects and underlying molecular mechanisms remain unclear. Purpose This study aimed to elucidate the protective mechanisms of combined GJ-PD against CIRI, with particular emphasis on mitochondrial transfer and neuronal PANoptosis. Methods Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to identify the chemical constituents of GJ-PD in brain. The mechanisms of GJ-PD in CIRI were investigated using transmission electron microscopy, Western blotting, immunofluorescence, immunohistochemistry, and pathological staining. In addition, an in vitro oxygen-glucose deprivation/reoxygenation (OGD/R)-induced co-culture injury model was established using HT22 neurons and C8-D1A astrocytes. Scanning electron microscopy and laser confocal microscopy combined with MitoTracker staining were applied to explore the effects of GJ-PD on mitochondrial transfer and neuronal PANoptosis. Furthermore, the involvement of CD38 and Miro1 was examined using CD38- and Miro1-overexpression plasmids, both alone and in combination with GJ-PD treatment. Results GJ-PD exhibited significant neuroprotective effects following CIRI. It reduced cerebral infarct volume, alleviated neuronal oxidative stress and mitochondrial dysfunction, and increased CD38 and Miro1 expression, thereby attenuating programmed cell death. These effects contributed to its anti-CIRI efficacy. In vitro results were consistent with in vivo findings. Treatment with GJ-PD-containing plasma alone in the co-culture system enhanced mitochondrial transfer from C8-D1A astrocytes to neurons via tunneling nanotubes (TNTs). This was accompanied by increased neuronal proliferation and ATP production, along with reduced neuronal PANoptosis. Co-treatment with CD38- or Miro1-overexpressing C8-D1A cells further improved cell viability and ATP levels, decreased the expression of programmed cell death-related proteins, and elevated CD38 and Miro1 protein expression. Notably, Miro1 overexpression did not significantly affect CD38 expression. Conclusions GJ-PD promotes the transfer of astrocytic mitochondria to neurons via TNTs through the CD38/Miro1 pathway. This process alleviates neuronal PANoptosis and improves neurological function, thereby exerting protective effects against CIRI.

Disulfidptosis is a pathologic process that occurs under conditions of NADPH deficiency and excess disulfide bonds in cells that express high levels of SLC7A11. This process is caused by glucose deprivation-induced disulfide stress and was first described by cancer researchers. Oxidative stress is a hypothesized mechanism underlying diseases of the central nervous system (CNS), and disulfide stress is a specific type of oxidative stress. Proteins linked to disulfidptosis and metabolic pathways involved in disulfidptosis are significantly associated with diseases of the CNS (neurodegenerative disease, neurogliomas and ischemic stroke). However, the specific mechanism responsible for this correlation remains unknown. This review provides a comprehensive overview of the current knowledge regarding the origin elements, genetic factors, and signaling proteins involved in the pathogenesis of disulfidptosis. It demonstrates that the disruption of thiometabolism and disulfide stress play critical roles in CNS diseases, which are associated with the potential role of disulfidptosis. We also summarize disulfidptosis-related drugs and highlight potential therapeutic strategies for treating CNS diseases. Additionally, this paper suggests a testable hypothesis that might be a promising target for treating CNS diseases.

Blood heat syndrome,one of the main subtypes of blood syndrome in traditional Chinese medicine(TCM),is mainly diagnosed by bleeding and heat manifestations and treated by the blood-cooling method.The biological essence of blood heat syndrome has not been elucidated yet,and there is a lack of systematic research on the potential mechanisms underlying the blood-cooling method.The biological essence of blood heat syndrome is closely related to abnormal immune response,oxidative stress,coagulation dysfunction,endocrine disorders,abnormalities in energy metabolism and so on.Blood heat syndrome is common in autoimmune skin diseases(such as systemic lupus erythematosus,psoriasis,and purpura),central hyperthermia,infectious diseases(such as infectious mononucleosis and COVID-19),and hemorrhagic diseases in gynecology.As the primary clinical therapy for blood heat syndrome,blood-cooling TCM is usually combined with the TCM with effects of activating blood and resolving stasis,nourishing Yin,and extinguishing wind to play the role of cooling blood.The mechanisms of above therapies may be attributed to reducing inflammation,inhibiting oxidative stress,restoring the balance of blood coagulation and metabolism,regulating the secretion of sex hormones,and alleviating allergic reactions.This article systematically explores the biological essence of blood heat syndrome and elucidates the targets and underlying mechanism of the blood-cooling method,laying a scientific foundation for the clinical application of TCM in the prevention and treatment of diseases associated with blood heat syndrome.

Objective: NOD-like receptor protein 3 (NLRP3)-mediated pyroptosis is pivotal in the pathological development of cerebral ischemia/reperfusion injury (CIRI). Although previous research has shown that electroacupuncture (EA) can alleviate CIRI through sirtuin-1 (SIRT1), the mechanism has not been well elucidated. Our study aimed to clarify whether the neuroprotective functions of EA are related to the reduction in NLRP3-mediated pyroptosis through the SIRT1 pathway. Materials and Methods: Rats received daily pretreatment with EA for 5 consecutive days before undergoing middle cerebral artery occlusion surgery. The Longa score was used to assess neurologic function. Infarct volume and morphological alterations were analyzed using 2,3,5-triphenyltetrazolium chloride and hematoxylin and eosin staining. In addition, neuronal pyroptosis was identified by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling/caspase-1 and neuronal nuclear antigen/caspase-1 immunofluorescence double staining. Levels of expression of pyroptosis markers were assessed by Western blotting and enzyme-linked immunosorbent assay. Results: EA improved deficits in neurologic function and minimized cerebral infarct volume. Mechanistically, a number of neuronal pyroptotic cells and protein levels of NLRP3, apoptosis-associated speck-like protein containing a CARD, and gasdermin D in the cerebral cortex were markedly reduced by EA treatment, and conversely, SIRT1 levels were increased. Notably, the specific SIRT1 inhibitor, EX527, reversed the effects of EA. Conclusions: EA potentially exerts a neuroprotective effect against CIRI through the SIRT1 pathway in NLRP3-mediated pyroptosis.

Central nervous system (CNS) disorders pose a significant health challenge due to high mortality and irreversible neurological damage. Current therapies are limited by insufficient efficacy and side effects, highlighting the urgent need for new multi-target neuroprotective agents. Polysaccharides from traditional Chinese medicines (TCMPs) exhibit considerable potential for CNS treatments. They ameliorate disease through several mechanisms, including countering oxidative stress, neuroinflammation, and neuronal apoptosis, maintaining blood-brain barrier (BBB) integrity, and regulating the gut-brain axis. Clinical translation encounters significant obstacles: complex extraction methods yielding low quantities, difficulties in definitive structural characterization, and inherently limited BBB permeability, limiting bioavailability. This review consolidates advancements in TCMP classification, extraction/purification, structural analysis, metabolism, transport, delivery systems, neuroprotective mechanisms, and CNS disorders targets. Evidence suggests that TCMPs confer neuroprotection by regulating microglial polarization, reinstating mitochondrial function, and enhancing neurotrophic factor secretion. Future research priorities include meticulous structural characterization to clarify structure-activity correlations, developing efficient BBB-penetrating delivery platforms, and conducting interdisciplinary mechanistic studies to promote the clinical application of TCMPs for CNS disorders.

Intracerebral hemorrhage (ICH) is the most morbid of all stroke types with a high mortality and significant disability burden. Inhibiting ferroptosis after ICH will effectively treat ICH and improve prognosis. Esculetin is the main active ingredient of the Chinese medicine Cortex Fraxini, which has neuroprotective effects. 7-methylguanosine (m7G) methylation is a common RNA modification that regulates mRNA stability and translation, playing an important role in neural development and function, as well as in ferroptosis-related pathways. In this study, we used hemin-induced PC12 cells to investigate the impact of esculetin on ferroptosis and the involvement of m7G modification. Cell viability was measured by cell counting kit-8. Cell death was evaluated by measuring the levels of LDH release and PI-positive cells. Ferroptosis was assessed by measuring the levels of Fe2+, glutathione, lipid reactive oxygen species, and malondialdehyde. The underlying mechanism was investigated by quantitative real-time PCR, methylated RNA immunoprecipitation (MeRIP), and RIP. Results suggested that esculetin promoted cell viability, inhibited ferroptosis, and increased NUDT1-mediated m7G levels in hemin-induced PC12 cells. Besides, NUDT1 interacted with GPX4 and enhanced the mRNA stability of GPX4. Silencing of GPX4 abolished the inhibitory influences of NUDT1 overexpressing on ferroptosis in hemin-induced PC12 cells. The ICH mouse model revealed that esculetin treatment effectively reduced neurological deficits and inhibited ferroptosis. In conclusion, esculetin treatment inhibits ICH-induced ferroptosis by promoting NUDT1-mediated m7G methylation modification and enhancing GPX4 stability. This study contributes to understanding the mechanisms by which esculetin may mitigate ICH damage and may provide a new potential therapeutic target for ICH.

Central nervous system (CNS) diseases, a leading cause of global disability and mortality, encompass a wide range of brain disorders such as stroke, Alzheimer's disease, Parkinson's disease, and so on. These diseases are characterized by dynamic cellular heterogeneity and disrupted intercellular crosstalk, yet their molecular drivers remain incompletely resolved. Single-cell RNA sequencing (scRNA-seq) dissects transcriptional diversity at cellular resolution, while spatial transcriptomics (ST) maps niche-specific interactions within tissue architecture-complementary approaches that have revealed disease-associated subpopulations, neural-glial communication, and microenvironmental remodeling. However, standalone omics layers inadequately capture the genetic, epigenetic, and functional cascades underlying CNS pathologies. Here, we highlight the transformative potential of integrating scRNA-seq and ST with multiomic profiling to delineate spatially orchestrated molecular networks. Such multiomic convergence enables systematic deconstruction of molecular mechanisms and intercellular communication across disease progression. By correlating these signatures with clinical phenotypes, this strategy accelerates biomarker discovery, patient stratification, and therapeutic target identification. We further discuss challenges in data harmonization, subcellular spatial resolution, and computational scalability that must be addressed to realize personalized CNS medicine. This synthesis advocates for interdisciplinary frameworks to translate multiomic insights into mechanistically grounded diagnostics and therapies, ultimately bridging the gap between molecular discovery and precision clinical intervention.

PANoptosis is a novelly defined mode of programmed cell death that involves the activation of multiple cellular death pathways, including pyroptosis, apoptosis, and necroptosis, triggering robust inflammatory reactions. Autophagy is a crucial cellular process that maintains cellular homeostasis and protects cells from various stresses. PANoptosis and autophagy, both vital players in the intricate pathological progression of ischemic stroke (IS), a brain ailment governed by intricate cell death cascades, have garnered attention in recent years for their potential interplay. While mounting evidence hints at a crosstalk between these two processes in IS, the underlying mechanisms remain elusive. Therefore, this review delves into and dissects the intricate mechanisms that underpin the intersection of PANoptosis and autophagy in this devastating condition. In conclusion, the crosstalk between PANoptosis and autophagy in IS presents a promising target for the development of novel stroke therapies. Understanding the interplay between these two pathways offers a much-needed insight into the underlying mechanisms of IS and opens the possibility for new therapeutic strategies.