Cerebral ischemic stroke (CIS) is characterized by high morbidity, disability, and mortality, representing a major global public health challenge and imposing a substantial social and economic burden worldwide. Cerebral ischemia/reperfusion (I/R) triggers complex pathological cascades, resulting in secondary brain injury, thereby limiting the therapeutic efficacy of single-target interventions. Therefore, the development of novel neuroprotective strategies with multitarget pharmacological properties remains an important research focus. Hydroxysafflower yellow A (HSYA), the major bioactive component of safflower (Carthamus tinctorius L.), has demonstrated protective effects in multiple experimental models of CIS. Accumulating preclinical evidence indicates that HSYA mitigates ischemic brain injury through multiple pathways, including maintaining mitochondrial homeostasis, suppressing excitotoxicity and calcium overload, attenuating oxidative stress, inhibiting inflammatory responses, and promoting angiogenesis. However, current mechanistic evidence is predominantly derived from cellular and animal studies. Limited clinical investigations have explored the effects of HSYA-containing preparations or HSYA injection in ischemic stroke; however, current evidence remains insufficient to establish definitive clinical efficacy. The clinical efficacy, optimal dosing strategies, and long-term benefits of HSYA remain to be fully established. In addition, limited brain distribution and unfavorable pharmacokinetic properties represent important challenges for its further clinical translation. This review summarizes the current understanding of the pharmacological effects of HSYA during different phases of CIS. Furthermore, from the perspective of the stroke-heart syndrome, the potential therapeutic value of HSYA in brain-heart comorbidity is discussed. Collectively, this review provides a new perspective on the therapeutic potential and translational challenges of HSYA in CIS management.

While photodynamic therapy (PDT) shows promise for tumor treatment, its efficacy is often constrained by the immunosuppressive tumor microenvironment. Pyroptosis, a Gasdermin-mediated inflammatory programmed cell death, augments PDT by releasing inflammatory cytokines and damage-associated molecular patterns that trigger robust antitumor immunity. This review systematically outlines the fundamental principles of PDT and critically analyzes existing immunological limitations in tumor treatment, highlighting the immunostimulatory mechanisms of pyroptosis in overcoming these specific therapeutic barriers. In addition, we summarize the rational design principles and recent advances in organelle-targeted photosensitizers-including those targeted to the plasma membrane, mitochondria, lysosomes, Golgi apparatus, and endoplasmic reticulum-for the effective induction of pyroptosis. We further discuss the persisting challenges associated with employing organelle-targeted photosensitizer-induced pyroptosis in tumor therapy. This review provides a strategic framework and future perspectives for developing next-generation precision photo-immunotherapies that harness pyroptosis-PDT synergy.

Incontinence-associated dermatitis (IAD) is a prevalent inflammatory skin condition caused by prolonged exposure to urine and feces, particularly in elderly and critically ill patients. The pathogenesis of IAD is closely linked to chronic inflammation, which results in the excessive release of pro-inflammatory cytokines and skin barrier dysfunction. This study investigates the therapeutic potential of quercetin, a natural flavonoid with known anti-inflammatory and antioxidant properties, in a rat model of IAD. We found that quercetin (50–100 mg/kg) downregulated the expression and phosphorylation of IKKα/β and NF-κB by over 40

Background: Hypersecretion of airway mucus is a hallmark of allergic asthma (AAS), hindering therapeutic agent penetration and potentially leading to fatal outcomes. This study aimed to enhance mucus penetration and degradation to improve drug delivery and therapeutic efficacy in AAS.Methods: We developed a chiral mesoporous silica (mSiO2)-based nanocomposite system loaded with N-acetylcysteine (NAC) and coated with exosomes (l-mSiO2@NAC@EXO). A mouse model of allergic asthma (C57BL/6 mice) was used to evaluate drug delivery, mucus clearance, airway resistance, and inflammatory responses.Results: The nano-exosome system exhibited uniform morphology, appropriate size distribution (~164 nm), and positive expression of exosomal markers (CD9, CD63, and TSG101), confirming successful fabrication. Functionally, l-mSiO2@NAC@EXO significantly enhanced mucus penetration and degradation compared to control formulations. In vivo, treatment markedly reduced airway resistance, suppressed mucus secretion, and decreased inflammatory cytokines (IL-4, IL-5, IL-13) in bronchoalveolar lavage fluid.Conclusions: This study demonstrates that a chiral nano-exosome system can improve drug delivery and therapeutic outcomes in allergic asthma by facilitating mucus clearance, offering a promising strategy for AAS treatment and broader biomedical applications.

Multiple sclerosis (MS) is a chronic autoimmune disorder of the central nervous system (CNS), characterized by neuroinflammation, demyelination, and subsequent axonal degeneration. Astrocytes (ASTs) play a dual role in MS pathogenesis, adopting either neurotoxic A1 or neuroprotective A2 phenotypes, which respectively inhibit or promote myelin repair. Fasudil (FSD), a Rho kinase inhibitor, has demonstrated potential in modulating AST phenotypes and facilitating remyelination. In this study, we investigated whether FSD treatment transformed astrocytes from A1 to A2 phenotype in cuprizone-induced demyelinating mice. Our results showed that FSD markedly suppressed A1 AST activation while enhancing A2 AST activation. Concurrently, FSD decreased pro-inflammatory chemokines such as CXCL1 and CXCL10, and increased A2-associated neurotrophic factors including BDNF and FGF2. These alterations fostered a favorable microenvironment that inhibited inflammatory responses and supported oligodendrocyte precursor cell (OPCs) differentiation and maturation, thereby ameliorated CPZ-induced demyelination. The study further revealed that these phenotypic transformation was mediated through the inhibition of NF-κB and LCN2 signaling pathways. This work identifies FSD as a promising therapeutic candidate for inhibiting demyelination through the regulation of AST polarization via specific signaling mechanism.

Experimental autoimmune encephalomyelitis (EAE) is a pivotal animal model for multiple sclerosis (MS) research. This study provides a comprehensive bibliometric analysis of 9,435 EAE-related publications from the Web of Science Core Collection (2000–2024) to map the research landscape, identify trends, and highlight emerging frontiers. The analysis reveals a shift from initial rapid growth to a stabilized output, with the USA, Germany, and China as the leading contributors. Immunology and Neurosciences form the core disciplines, with increasing integration of cell biology, microbiology, and pharmacology. Co-citation and keyword analyses demonstrate a clear evolution in research focus: while early studies centered on T-cell biology and classic neuroinflammation, recent investigations emphasize the gut-brain-immune axis, microglial/astrocyte reactivity, immunometabolism, and novel therapeutic strategies like microbiota-targeted interventions and nanomedicine. The Th17/Treg axis remains a central immunoregulatory hub. This analysis delineates the intellectual structure of the field, underscoring its progression from foundational immune mechanisms to complex, system-level understandings of neuroimmune crosstalk, and identifies promising directions for future MS therapeutic development.Experimental autoimmune encephalomyelitis (EAE) is a pivotal animal model for multiple sclerosis (MS) research. This study provides a comprehensive bibliometric analysis of 9,435 EAE-related publications from the Web of Science Core Collection (2000–2024) to map the research landscape, identify trends, and highlight emerging frontiers. The analysis reveals a shift from initial rapid growth to a stabilized output, with the USA, Germany, and China as the leading contributors. Immunology and Neurosciences form the core disciplines, with increasing integration of cell biology, microbiology, and pharmacology. Co-citation and keyword analyses demonstrate a clear evolution in research focus: while early studies centered on T-cell biology and classic neuroinflammation, recent investigations emphasize the gut-brain-immune axis, microglial/astrocyte reactivity, immunometabolism, and novel therapeutic strategies like microbiota-targeted interventions and nanomedicine. The Th17/Treg axis remains a central immunoregulatory hub. This analysis delineates the intellectual structure of the field, underscoring its progression from foundational immune mechanisms to complex, system-level understandings of neuroimmune crosstalk, and identifies promising directions for future MS therapeutic development.

Ischemic stroke is a significant cause of disability and mortality on a global scale, with neuronal dysfunction playing a critical role in its pathogenesis. Conventional treatment approaches for ischemic stroke involve surgical interventions and thrombolytic therapy, yet these methods frequently result in ischemia/reperfusion (I/R) injury. Recent studies have underscored the implication of diverse programmed cell death mechanisms, including ferroptosis, in the progression of ischemic stroke. Ferroptosis, a newly recognized form of cell death reliant on iron, is intricately linked to various neurological conditions. Despite the existing body of research on ferritinophagy and neuronal ferroptosis in the context of cerebral ischemia-reperfusion injury, there is a lack of understanding regarding the mechanisms involved in neuronal ferroptosis. This study seeks to explore the relationship between neuronal autophagy and neuronal ferroptosis using in vivo and in vitro models of cerebral ischemia/reperfusion. The findings of our study reveal a significant upregulation of the ferritinophagy-associated protein NCOA4 following cerebral ischemia/reperfusion, concomitant with the initiation of ferroptosis in neuronal cells. This observation offers compelling support for a direct association between neuronal ferritinophagy and ferroptosis. Hydroxysafflor Yellow A (HSYA), a traditional Chinese herb, shows promise in reducing brain ischemia/reperfusion injury, but its exact protective mechanism is still unknown. Our study reveals a new way HSYA protects the brain by preventing neuronal ferroptosis after a stroke, a mechanism not previously reported.

BACKGROUND:High blood glucose is a well-established risk factor for poor outcomes in ischemic stroke. However, the underlying molecular mechanisms linking high blood glucose to worsened stroke outcomes remain unclear. OBJECTIVES:Previous studies have implicated the NLRP3 inflammasome, a key mediator of neuroinflammation, in cerebral ischemia/reperfusion (I/R) injury. Under high blood glucose conditions, NLRP3 activation is amplified, potentially driving a vicious cycle of inflammation and neuronal death. Yet, how high blood glucose specifically modulates NLRP3 activation and its downstream pathways remains unclear. This study aimed to investigate the specific mechanisms by which high glucose enhances NLRP3 inflammasome activity and contributes to worsened brain injury following cerebral I/R. METHODS:We employed a combination of in vitro and in vivo experimental approaches to explore the impact of high glucose on NLRP3 inflammasome activation and its consequences on ischemic stroke outcomes. In vitro experiments were conducted by culturing various immune cells in high-glucose conditions to evaluate the activation of the NLRP3 inflammasome and the mitochondrial association of HK2. In vivo, mice with genetic knockouts of Nlrp3, Pycard (the gene encoding ASC), or microglial-specific Hk2 were subjected to transient middle cerebral artery occlusion (tMCAO). RESULTS:Our findings revealed that the activation of the NLRP3 inflammasome was enhanced post cerebral I/R under high glucose and a N-terminal truncation of NLRP3 (miniNLRP3) was induced. Overexpression of PKA could promote the generation of miniNLRP3, while inhibition of PKA decreased the generation of miniNLRP3. In addition, treatment with pan serine protease could block PKA and LPS mediated generation of miniNLRP3. Overexpression of the N-terminal truncation of NLRP3 could potentiate the activation of the NLRP3 inflammasome under high glucose conditions by promoting the dissociation of Hexokinase 2 (HK2) from mitochondria. In addition, knockout of Nlrp3, Pycard, or microglial Hk2, could all attenuate cerebral I/R-induced brain injury under high blood glucose in mice. CONCLUSION:Our study elucidates PKA-mediated generation of a 30 kD N-terminal truncation of NLRP3 (miniNLRP3) in a serine protease-dependent manner, which could potentiate the activation of the NLRP3 inflammasome under high glucose conditions via promoting the dissociation of HK2 from mitochondria. These findings add a new dimension to our understanding of NLRP3 regulation in the context of stroke injury, and suggest that the PKA-miniNLRP3-HK2-NLRP3 pathway is a promising therapeutic strategy to improve stroke outcomes in patients with elevated blood glucose levels.

This study aims to explore the effects and action mechanisms of the active ingredients in Buyang Huanwu Decoction(BYHWD), namely tetramethylpyrazine(TMP) and hydroxy-safflor yellow A(HSYA), on oxygen-glucose deprivation/reglucose-reoxygenation(OGD/R)-induced inflammation and oxidative stress of microglia(MG). Network pharmacology was used to screen the effective monomer ingredients of BYHWD and determine the safe concentration range for each component. Inflammation and oxidative stress models were established to further screen the best ingredient combination and optimal concentration ratio with the most effective anti-inflammatory and antioxidant effects. OGD/R BV2 cell models were constructed, and BV2 cells in the logarithmic growth phase were divided into a normal group, a model group, an HSYA group, a TMP group, and an HSYA + TMP group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines such as interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6). Oxidative stress markers, including superoxide dismutase(SOD), nitric oxide(NO), and malondialdehyde(MDA), were also measured. Western blot was used to analyze the protein expression of both inflammation-related pathway [Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)] and oxidative stress-related pathway [nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)]. Immunofluorescence was used to assess the expression of proteins such as inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). The most effective ingredients for anti-inflammatory and antioxidant effects in BYHWD were TMP and HSYA. Compared to the normal group, the model group showed significantly increased levels of IL-1β, TNF-α, IL-6, NO, and MDA, along with significantly higher protein expression of NF-κB, TLR4, Nrf2, and HO-1 and significantly lower SOD levels. The differences between the two groups were statistically significant. Compared to the model group, both the HSYA group and the TMP group showed significantly reduced levels of IL-1β, TNF-α, IL-6, NO, and MDA, lower expression of NF-κB and TLR4 proteins, higher levels of SOD, and significantly increased protein expression of Nrf2 and HO-1. Additionally, the expression of the M1-type MG marker iNOS was significantly reduced, while the expression of the M2-type MG marker Arg-1 was significantly increased. The results of the HSYA group and the TMP group had statistically significant differences from those of the model group. Compared to the HSYA group and the TMP group, the HSYA + TMP group showed further significant reductions in IL-1β, TNF-α, IL-6, NO, and MDA levels, along with significant reductions in NF-κB and TLR4 protein expression, an increase in SOD levels, and elevated Nrf2 and HO-1 protein expression. Additionally, the expression of the M1-type MG marker iNOS was reduced, while the M2-type MG marker Arg-1 expression increased significantly in the HSYA + TMP group compared to the TMP or HSYA group. The differences in the results were statistically significant between the HSYA + TMP group and the TMP or HSYA group. The findings indicated that the combined use of HSYA and TMP, the active ingredients of BYHWD, can effectively inhibit OGD/R-induced inflammation and oxidative stress of MG, showing superior effects compared to the individual use of either component.

The addition of microstructures to the inner surface of the stent reduces resistance and inhibits the phenomenon of blood adhesion. In this study, the design of a fish-scale microstructured vascular stent was proposed based on bionics, and its main design parameters were optimized using the finite element method. In addition, the hemodynamic effects of a standard stent and a fish-scale microstructured stent on an ideal cerebral aneurysm were comparatively analyzed. The results showed that the fish-scale microstructured stent significantly accelerated intraluminal blood flow velocity by 11.6% compared to the standard stent. In addition, the fish-scale microstructured stent was able to reduce blood flow into the aneurysm lumen by 28.6%.

Introduction:Neuroinflammation induced by astrocytes has garnered significant attention recently. The IL-17A/IL-17RA signal pathway plays an important role in ischemic stroke (IS). Hydroxysafflor yellow A (HSYA) has been reported to have anti-apoptotic and anti-inflammatory properties that can protect neurons. In this study, we explore a novel mechanism that underlies the anti-apoptotic and anti-inflammatory effects of HSYA. Methods:In vitro experiments were carried out using primary astrocytes and HT22 neuronal cells in an oxygen-glucose deprivation/reoxygenation (OGD/R) model. Techniques such as Western blot, immunofluorescence staining, Enzyme-linked immunosorbent assay (ELISA), and quantitative real-time polymerase chain reaction (qRT-PCR) were utilized to detect relevant indicators. The purpose was to investigate the effect of HSYA on the influence of IL-17A secreted by primary astrocytes after OGD/R on HT22 neuronal cells. Results:The results indicated that the production of IL-17A by astrocytes increased following OGD/R, which was reduced due to HSYA treatment. In addition, astrocyte-derived IL-17A resulted in neuronal cell damage. Further studies showed that HSYA reduced IL-17A production by inhibiting activation of the IL-17RA/ACT1/NF-κB/IL-17A loop, which ultimately alleviated neuroinflammation and reduced neuronal apoptosis. Discussion:These findings suggest that an activated loop indeed exists between IL-17A and IL-17RA/ACT1/NF-κB after OGD/R, and HSYA treatment alleviated IL-17A release from astrocytes after OGD/R by inhibiting the IL-17RA/ACT1/NF-κB/IL-17A loop. These results further emphasize the anti-inflammatory and neuroprotective effects of HSYA and suggest that it may be a promising drug for treating IS.

ObjectiveThe aim of this study was to observe the clinical efficacy and safety of conventional treatment combined with acupuncture in the treatment of idiopathic deafness.MethodsThis retrospective study enrolled 141 patients with idiopathic deafness hospitalized from January 2018 to December 2019. We compared the patients according to previous therapy (conventional treatment and acupuncture treatment), demographics, type of deafness, pure tone hearing threshold score, concomitant symptoms, adverse events and outcomes.ResultsA total of 141 patients were assigned to a control group (n = 67, with simple routine Western medicine treatment) and an observation group (n = 74, with acupuncture treatment on the basis of Western medicine therapy). The total effective rate was 98.65% in the observation group, compared with 74.63% in the control group (p < 0.001). The tinnitus and vertigo were both improved in both groups, and the observation group was better than the control group (p < 0.05). Adverse reactions occurred in one patient (1.35%) in the observation group and in five patients (7.46%) in the control group (p = 0.085).ConclusionAmong patients with idiopathic deafness who can be treated after the onset of symptoms, the combination of acupuncture and conventional treatment is superior to conventional treatment alone for improving clinical efficacy, reducing concomitant symptoms and not increasing the risk of adverse events.

Oxidative stress and mitochondrial dynamics imbalance are key contributors to AD pathogenesis. GLPS, an extract from Ganoderma lucidum spores, exhibits anti-inflammatory, antioxidant, and immunomodulatory properties. However, the roles of GLPS in regulating oxidative stress and mitochondrial dynamics in AD remain poorly understood. Here, the underlying mechanisms of neuroprotective effects on cognitive dysfunction in 5 × FAD mice were explored. C57BL/6 mice served as WT controls, while 5 × FAD mice were divided into an AD group and an AD + GLPS group. The mice in AD + GLPS group were administered daily GLPS (25 mg/kg) by i.p. injection for two months, while WT and AD mice received an equivalent volume of normal saline. The results indicated that GLPS markedly improved cognitive function and decreased p-tau and Aβ levels in 5 × FAD mice. Moreover, GLPS alleviated oxidative stress by increasing SOD levels and decreasing MDA concentrations. It also inhibited excessive mitochondrial fragmentation by decreasing the expression of p-Drp1 and Fis1, while increasing the levels of Mfn1, Mfn2, and OPA1 in 5 × FAD mice. Mechanistically, GLPS activated Nrf2, leading to a marked upregulation of antioxidant enzymes, including HO- 1, NQO1, and SOD2 in 5 × FAD mice. Collectively, these findings suggest that GLPS ameliorates cognitive deficits in 5 × FAD mice by reducing oxidative stress and modulating mitochondrial dynamics through Nrf2-mediated antioxidant enzyme activation.

Lipocalin-2 (LCN2), an acute phase protein mainly expressed in astrocytes (Ast), is closely related to the production of inflammatory cytokines following ischemic stroke. During the pathophysiological process of ischemic stroke, the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway is activated. Despite evidence suggesting some link between the two, the relationship between the JAK2/STAT3 signaling pathway and the LCN2 expression in Ast following brain ischemia is incompletely understood. Hydroxysafflower yellow A (HSYA), an active ingredient found in Carthamus tinctorius L flowers, has been demonstrated to effectively mitigate cerebral ischemia via its anti-inflammatory effect. However, whether HSYA mitigates the neuroinflammatory damage after ischemic stroke by disrupting the interaction between the JAK2/STAT3 signaling pathway and LCN2 in Ast is unknown. Focusing on these two scientific questions, we established an in vivo middle cerebral artery occlusion/reperfusion (MCAO/R) rat model and in vitro primary astrocyte oxygen glucose deprivation/reperfusion (OGD/R) model. In vivo results showed that HSYA treatment alleviated nerve damage and inhibited the expression of LCN2 and inflammatory factors in Ast. In vitro results showed after OGD/R the expression of LCN2 and inflammatory cytokines increased and the JAK2/STAT3 was activated in Ast. Meanwhile, after OGD/R the JAK2/STAT3 activation in Ast increased LCN2 expression, and the inhibition of LCN2 expression by HSYA decreased the JAK2/STAT3 activation in Ast. These findings suggest that there is an interaction between the LCN2 and JAK2/STAT3 in Ast after ischemic stroke, which can enhance the inflammatory factors and exacerbate neuroinflammatory injury. Therefore, we conclude that HSYA may inhibit the LCN2/STAT3 loop in Ast, thereby mitigating neuroinflammation after cerebral ischemia.

BACKGROUND:Depression is the most common neuropsychiatric sequela of stroke, negatively affecting functional recovery and quality of life. Although virtual reality-based rehabilitation (VRBR) is an effective intervention in stroke rehabilitation, its efficacy in treating poststroke depression (PSD) remains unclear. OBJECTIVES:To evaluate the treatment effect of VRBR on PSD. DATA SOURCES AND METHODS:Six medical databases were systematically searched from inception to May 31, 2024, with an update on October 5, 2024. Potentially eligible studies were screened against the inclusion and exclusion criteria. Review Manager 5.4 and Stata 14.0 were used to conduct statistical analysis. RESULTS:Twelve studies published between 2015 and 2024 were included. The pooled results revealed that VRBR significantly reduced depression scores among poststroke patients [SMD = -0.56, 95 % CI (-0.90,-0.23), P = 0.001]. Subgroup analyses suggested numerically greater improvements in patients with a mean age < 60 years and those receiving ≥20 intervention sessions, though between-subgroup differences were not statistically significant. CONCLUSION:VRBR may be a promising therapy for PSD. However, methodologically rigorous studies are needed to further confirm its efficacy and safety. TRIAL REGISTRATION:PROSPERO, CRD42024550019.

While cancer immunotherapy has achieved groundbreaking clinical success, its efficacy is frequently compromised by insufficient T-cell activation, the immunosuppressive tumor microenvironment (TME), and off-target toxicity. Pyroptosis, a highly immunogenic form of programmed cell death characterized by gasdermin-mediated pore formation, massive cytokine release (e.g., IL-1β and IL-18), and robust dendritic cell activation, offers a compelling strategy to overcome these limitations. This review critically examines how nanotechnology-enabled pyroptosis induction can potentiate immunotherapy by (1) classifying pyroptosis-inducing nanomaterials into five combinatorial therapeutic platforms - immune checkpoint inhibitors, vaccine adjuvants, oncolytic virus-coupled systems, innate immune sensitizers, and multi-modal hybrids; (2) elucidating their mechanisms in reshaping the TME via pyroptosis-induced immunogenicity and bystander immune cell activation; and (3) highlighting unresolved challenges, including tumor-intrinsic pyroptosis resistance, nanoparticle biodistribution barriers, and cytokine storm risks. By integrating fundamental insights with translational perspectives, this work provides a strategic framework for developing pyroptosis-nanotechnology synergies to achieve precision immune modulation.

Buyang Huanwu Decoction(BYHWD),as one of the classic formulas in traditional Chinese medicine(TCM)for the treatment of cerebral ischemic stroke(CIS),has demonstrated definite effects in clinical practice.However,the material basis and mechanism of treatment have not been systematically elucidated.This study employed network pharmacology and molecular docking to analyze the potential targets and mechanisms of blood-and brain-penetrating active components of BYHWD in reducing cell apoptosis in CIS.Cell experiments were then carried out to validate the prediction results.In the experiments,five active components including hydroxysafflor yellow A(HSYA),tetramethylpyrazine(TMP),astragaloside Ⅳ(AS-Ⅳ),amygdalin(AMY),and paeoniflorin(PF)were selected to explore the pharmacological effects of BYHWD.HT22 cells were treated with BYHWD,and the cell counting kit-8(CCK-8)method was employed to examine the toxic and side effects of BYHWD.A cell model of oxygen-glucose deprivation/reoxygenation(OGD/R)was constructed,with apoptosis and pyroptosis as the main screening indicators.The levels of lactate dehydrogenase(LDH)and glutathione(GSH)were measured to assess the cell membrane integrity.Flow cytometry was employed to detect apoptosis,and the activities of caspase-3 and caspase-1 were measured to clarify the status of apoptosis and pyroptosis.ELISA was employed to determine the levels of interleukin(IL)-1β and IL-18 to confirm pyroptosis.HSYA and AMY were identified in this study as the active components regulating apoptosis and pyroptosis.TUNEL was employed to detect the apoptosis rate,and Western blot was employed to determine the expression levels of apoptosis-related proteins B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and caspase-3,which confirmed that the anti-apoptotic effect of the combined component group was superior to that of the single component groups.The molecular docking results revealed strong binding affinity of HSYA and AMY with SDF-1α and CXCR4.AMD3100,a selective antagonist of CXCR4,was then used for intervention.The results of Western blot showed alterations in the expression levels of apoptosis-associated proteins,SDF-1α,and CXCR4.In conclusion,HSYA and AMY influence cellular apoptosis by modulating the SDF-1α/CXCR4 signaling cascade.

The treatment of immunomodulation in multiple sclerosis (MS) can alleviate the severity and relapses. However, it cannot improve the neurological disability of patients due to a lack of myelin protection and regeneration. Therefore, remyelinating therapies may be one of the feasible strategies that can prevent axonal degeneration and restore neurological disability. Natural product icariin (ICA) is a flavonol compound extracted from epimedium flavonoids, which has neuroprotective effects in several models of neurological diseases. Here, we attempt to explore whether ICA has the potential to treat demyelination and its possible mechanisms of action using lipopolysaccharide-treated BV2 microglia, primary microglia, bone marrow-derived macrophages, and cuprizone-induced demyelination model. The indicators of oxidative stress and inflammatory response were evaluated using commercial kits. The results showed that ICA significantly reduced the levels of oxidative intermediates nitric oxide, hydrogen peroxide, malondialdehyde, and inflammatory cytokines TNF-α, IL-1β, and increased the levels of antioxidants superoxide dismutase, catalase, glutathione peroxidase, and anti-inflammatory cytokines IL-10 and TGF-β in vitro cell experiments. In vivo demyelination model, ICA significantly alleviated the behavioral abnormalities and enhanced the integrated optical density/mm 2 of Black Gold II and myelin basic protein myelin staining, accompanied by the inhibition of oxidative stress/inflammatory response. Immunohistochemical staining showed that ICA significantly induced the expression of nuclear factor erythroid derived 2/heme oxygenase-1 (Nrf2/HO-1) and inhibited the expression of toll-like receptor 4/ nuclear factor kappa B (TLR4/NF-κB), which are two key signaling pathways in antioxidant and anti-inflammatory processes. Our results strongly suggest that ICA may be used as a potential agent to treat demyelination via regulating Nrf2/HO-1-mediated antioxidative stress and TLR4/NF-κB-mediated inflammatory responses.