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.

Autoimmune Hepatitis (AIH) and Multiple Sclerosis (MS) are chronic inflammatory diseases with abnormal immune responses. This study aims to identify common biomarkers for AIH and MS using bioinformatics analysis. Gene expression data of AIH (GSE159676) and MS (GSE131279 and GSE131281) were obtained from the GEO database. Differentially Expressed Genes (DEGs) were identified using the limma package in R. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, Protein-Protein Interaction (PPI) network, and machine learning algorithm Least Absolute Shrinkage and Selection Operator (LASSO) regression were used to evaluate potential biomarkers. The common biomarker gene Zinc Finger Protein Y-linked (ZFY) was identified. KEGG analysis showed significant enrichment of the Phosphatidylinositol 3-Kinase/Protein Kinase B (PI3K/Akt) pathway in both diseases. LASSO regression identified ZFY as a potential diagnostic marker, with decreased expression in both AIH and MS groups. Single-gene immune infiltration analysis indicated a significant association between ZFY expression and immune cell infiltration levels. Experimental validation in ConA-induced hepatitis and CPZ-mediated demyelination model mice further verified the diagnostic potential of ZFY. This study reveals the potential of ZFY as a biomarker for AIH and MS, highlighting its role in the PI3K/Akt pathway and immune infiltration. These findings provide new insights into the common pathological mechanisms of AIH and MS and suggest potential targets for future therapeutic strategies.

Objective:To observe the effects of electroacupuncture (EA) on high mobility group box-1 (HMGB1)/ receptor for advanced glycation end products (RAGE)/nuclear factor kappa-B (NF-κB) pathway-mediated neuroinflammatory response and reactive astrocyte in Parkinson's disease (PD) mice, and to explore the mechanism of EA in the prevention and treatment of PD. Methods:Thirty-six male C57BL/6 mice were randomly divided into a control group, a model group, and an EA group, with 12 mice in each group. The PD model was established by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days. From the first day of model establishment, mice in the EA group received EA at "Baihui" (GV20) and bilateral "Shenshu" (BL23), with continuous wave, frequency of 2 Hz and intensity of 2 mA, 15 min each time, once daily, for 14 days. Pole test, hanging test, and gait analysis were used to assess behavioral performance. Immunofluorescence staining was used to detect tyrosine hydroxylase (TH) and glial fibrillary acidic protein (GFAP) positive cells in the substantia nigra of the midbrain. ELISA was used to detect α-synuclein (α-syn) content in the substantia nigra. Western blot was used to detect protein expression levels of TH, HMGB1, RAGE, NF-κB, phosphorylated NF-κB (p-NF-κB), GFAP, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) in the substantia nigra. Real-time quantitative PCR was used to detect mRNA expression levels of HMGB1, RAGE, NF-κB, GFAP, TNF-α, IL-6, and IL-10 in the substantia nigra. Results:Compared with the control group, the model group showed prolonged pole test time (P<0.01), decreased hanging score (P<0.01); shortened stride length and standing time (P<0.01), increased step frequency (P<0.01), and prolonged swing time (P<0.01) of bilateral forelimbs and hindlimbs; the number of TH-positive cells and TH protein expression level, as well as IL-10 protein and mRNA expression levels in the substantia nigra were decreased (P<0.01, P<0.05), while α-syn content and the number of GFAP-positive cells, protein and mRNA expression levels of HMGB1, RAGE, GFAP, TNF-α, and IL-6, as well as p-NF-κB/NF-κB and NF-κB mRNA expression were increased (P<0.05, P<0.01). Compared with the model group, the EA group showed shortened pole test time (P<0.01), increased hanging score (P<0.05); increased stride length (P<0.05, P<0.01), decreased step frequency (P<0.01), prolonged standing time (P<0.05, P<0.01), and shortened swing time (P<0.05) of bilateral forelimbs and hindlimbs; the number of TH-positive cells, TH protein level, and IL-10 protein and mRNA expression levels in the substantia nigra were increased (P<0.01, P<0.05), while α-syn content and the number of GFAP-positive cells, protein and mRNA expression levels of HMGB1, RAGE, GFAP, TNF-α, and IL-6, as well as p-NF-κB/NF-κB and NF-κB mRNA expression were decreased (P<0.05, P<0.01). Conclusion:EA can improve motor dysfunction in PD mice, protect dopaminergic (DA) neurons, and reduce α-syn protein aggregation, thereby exerting a neuroprotective effect. This effect may be related to inhibition of reactive astrocyte activation and the HMGB1/RAGE/NF-κB pathway, thereby reducing neuroinflammatory responses.

Background and Purpose Neuroinflammation, characterized by dysregulated activation of microglia, is a hallmark of Parkinson's disease (PD). Nevertheless, therapeutic strategies aimed at the mechanism of inflammation resolution remain limited. Study design and Methods This study integrated PD clinical cohorts, MPTP-induced and miR-146a-induced mouse models, as well as LPS-stimulated and miR-146a-activated cell models. Combined with omics analysis, behavioral detection and molecular biology experiments, we systematically evaluated the anti-inflammatory protective effects of Wuzi Yanzong Pills (WYP). The active plant metabolites of WYP were identified using a combination of UHPLC-Q-Exactive-MS/MS, AP-SMALDI Orbitrap MSI, and pharmacokinetic analysis. Results In PD patients, WYP significantly improved motor dysfunction, inhibited pro-inflammatory cytokines, and elevated neurotransmitter levels. Exosomal miRNA sequencing analysis indicated that miR-146a-5p may serve as a biomarker for PD and is positively correlated with disease severity. Animal experiments further showed that WYP improved motor symptoms and neuroinflammation in MPTP- and miR-146a-induced PD mice models. A dual-luciferase reporter assay confirmed ubiquitin specific peptidase 3 (USP3) as a direct target gene of miR-146a-5p. In an LPS-activated BV2 microglial cell model, WYP intervention reduced the content of miR-146a-5p in cell-derived exosomes and mitigated their pro-inflammatory damaging effects on neuronal cells. Component analysis revealed that 16 plant metabolites in WYP can enter the bloodstream, among which 11 can cross into the brain. Notably, geniposidic acid, hyperoside, kaempferol, protocatechuic acid, and schisandrol A significantly suppressed the expression of pro-inflammatory factors in BV2 cells, suggesting that they may be the main active components underlying the anti-inflammatory effects of WYP. Conclusion WYP improves PD by regulating the miR-146a-5p/USP3/NF-κB pathway. Meanwhile, the active plant metabolites of WYP have been identified. These findings provide experimental evidence for WYP as a potential therapeutic agent for PD.

This study compares the synthesis mechanism, structural characteristics, and anti-neuroinflammatory activity of Astragalus polysaccharides (APS) from four parts of Astragalus membranaceus: root, stem, leaf, and flower. Ultrasonic-assisted extraction (UAE) was used to determine the optimal extraction conditions for APS: ultrasonic power of 291.76 watts, extraction time of 61.45 min, solid-liquid ratio of 1:27.88, and temperature of 60 °C, which significantly increased the APS yield. The highest APS content was found in the root (16.34 %), followed by the stem and leaf, with the flower containing the least. Transcriptomic and miRNA sequencing revealed significant differences in gene expression among tissues. Genes related to APS synthesis were most active in the root, while the stem, leaf, and flower exhibited distinct expression profiles related to secondary metabolite synthesis. Functional experiments demonstrated that APS from all parts inhibited LPS-induced inflammatory responses in BV2 microglial cells, reducing the production of NO, TNF-α, and IL-1β, with root-derived APS showing the strongest activity. Structural analyses further indicated notable differences in monosaccharide composition, molecular weight, and ultrastructure among APS from different parts. Root APS was predominantly composed of glucose (91.01 %) and had the lowest molecular weight. Flower APS had the highest molecular weight and was rich in galacturonic acid and galactose. Leaf APS contained rhamnose (22.76 %) and displayed unique structural features. These findings provide multi-dimensional insights into the tissue-specific regulatory mechanisms and bioactivity variations of APS, offering a theoretical foundation for the targeted development and utilization of Astragalus polysaccharides.

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.

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.

Astragaloside IV (AS-IV) is a bioactive compound derived from Radix Astragali, a traditional Chinese herb widely used as a dietary supplement to enhance immune function. Modern pharmacological studies have demonstrated that AS-IV exhibits anti-inflammatory and immunomodulatory properties. In this study, we investigated the effects of AS-IV on motor dysfunction, microglial polarization, and immune regulation mechanisms in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) mouse model. Our results showed that AS-IV (40 mg/kg) significantly improved motor function in PD mice, as evidenced by reduced descent time in the pole test, increased hanging score in the hanging test, increased stride lengths, and reduced paw angle in the gait test. Furthermore, AS-IV administration attenuated the loss of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra pars compacta (SNpc), promoted microglial polarization from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype, suppressed the levels of pro-inflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNFα), enhanced the levels of anti-inflammatory cytokines including interleukin-4 (IL-4) and interleukin-10 (IL-10) in the SNpc of PD mice. Mechanistically, AS-IV significantly downregulated the expression and phosphorylation levels of TLR4/p38 (JNK)/NF-κB pathway-related proteins, including Toll-like receptor 4 (TLR4), Myeloid differentiation primary response protein 88 (MyD88), Apoptosis signal-regulating kinase 1 (ASK1), Mitogen-activated protein kinase 3/6 (MKK3/6), Phosphorylated-MKK3/6 (p-MKK3/6), Phosphorylated-mitogen-activated protein kinase 4/7 (p-MKK4/7), p38 mitogen-activated protein kinase (p38), Phosphorylated-p38 (p-p38), c-Jun N-terminal kinase (JNK), Phosphorylated-JNK (p-JNK), nuclear factor kappa-B (NF-κB), and Phosphorylated-NF-κB (p-NF-κB). To further validate the targeting effect of AS-IV, 1 mg/kg of LPS-EB Ultrapure was utilized as a specific TLR4 agonistwe to selectively activated the TLR4/NF-κB signaling pathway without triggering other inflammatory pathways, leading to elevated mRNA levels of TLR4, NF-κB, IL-1β, IL-6, TNFα and protein expression of TLR4, p-JNK, p-p38, p-NF-κB, IL-1β, IL-6, TNFα in the SNpc of PD mice. Importantly, AS-IV pretreatment can't counteract these LPS-EB Ultrapure-triggered effects, demonstrating its dependence on the TLR4/NF-κB signaling pathway. In conclusion, our findings indicate that AS-IV modulates microglial polarization and attenuates neuroinflammation by inhibiting the TLR4/NF-κB pathway, thereby ameliorating motor dysfunction and neuronal loss in PD mice.

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.

Bone marrow-derived mesenchymal stem cells (BMSCs) are promising candidates for treating autoimmune diseases like multiple sclerosis (MS) due to their ability to differentiate into multiple lineages and their immunomodulatory properties. However, the immunomodulatory capacity of BMSCs is highly adaptable, and primarily regulated by inflammatory factors. In this study, we evaluated the therapeutic effectiveness of BMSCs in dexamethasone (DEX)-pretreated experimental autoimmune encephalomyelitis (EAE) mice. Our results demonstrated a significant interaction between DEX and BMSCs. In contrast to their effect in non-pretreated mice, BMSCs administration in DEX-pretreated EAE mice resulted in a significant increase in infiltrating CD4+ T cells and a concomitant decrease in regulatory T (Treg) cell populations in the central nerves system, which likely resulted from DEX-induced changes in the peripheral immune microenvironment. Our findings in the EAE model indicate that the immune microenvironment established by DEX pretreatment is detrimental to the efficacy of BMSC therapy. This preclinical evidence suggests that evaluating the peripheral immune status may be a critical consideration for future clinical studies of MSC treatment in MS.

Background Traditional Chinese medicine offers unique and valuable resources for the exploration of novel approaches to disease treatment. Panaxadiol, an active compound derived from the transformation of ginsenosides. It has a variety of pharmacological activities. However, there is a lack of research regarding its effects on Parkinson's disease (PD). Purpose and Study design This study investigates the activity and mechanism of panaxadiol in the treatment of PD both in vivo and in vitro. In addition, a new method has been developed to extract panaxadiol. Results The results showed that in vitro, panaxadiol can reduce the viability and cytotoxicity of BV2 cells induced by LPS, and inhibit the production of pro-inflammatory factors through the TLR4/MyD88/NF-κB pathway. In vivo, panaxadiol can improve motor and intestinal dysfunction in PD mice and repair the loss of DA neurons. Further studies have shown that panaxadiol treatment alleviates damage to the blood-brain barrier (BBB), inhibits neuroinflammation in the substantia nigra (SN), and further reduces damage to DA neurons. Subsequently, it was found that panaxadiol alleviated peripheral inflammation and significantly restored the intestinal microbial community. Further mechanistic studies found that panaxadiol treatment inhibited the TLR4/MyD88/NF-κB signaling pathway and its downstream pro-inflammatory products in the SN. In addition, Additionally, the response surface method was employed to identify the optimal conditions for extracting panaxadiol using a deep eutectic solvent (NADES). Conclusion In summary, this study has found for the first time that panaxadiol can effectively improve the symptoms of PD by regulating neuroinflammation, repairing the BBB, and gut microbiota. Meanwhile, adopting a green extraction process significantly increases the content of panaxadiol, providing a new direction for the development of PD candidate drugs.

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.

This study employed molecular simulation docking technology to identify the most suitable host molecules for encapsulating Astragaloside IV (AS-IV). AS-IV-HP-β-CD molecular capsules were prepared by encapsulating the hydrophobic groups of AS-IV in the inner cavity of HP-β-CD through the grinding method. The encapsulation behaviours were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, and liquid nuclear magnetic resonance spectroscopy. It was found that AS-IV was encapsulated inside the HP-β-CD cavity and presented a layered structure after encapsulation. Content determination showed that the solubility and dissolution rate of AS-IV-HP-β-CD in vitro were significantly increased compared to AS-IV. Pharmacodynamic tests demonstrated that oral administration of AS-IV-HP-β-CD effectively ameliorated motor dysfunction, alleviated dopaminergic (DA) TH-positive neuronal loss, and reduced the expression of inflammatory cytokines IL-1β and IL-18 in the substantia nigra pars compacta of MPTP-induced Parkinson's disease mice. Simultaneously, the AS-IV-HP-β-CD intervention inhibited the expression of the NLRP3 inflammasome, accompanied by a decrease in the expression of TLR4/MyD88/NF-κB pathway proteins. This study addressed the issues of poor water solubility and dissolution rate, laying the groundwork for developing AS-IV formulations. Meanwhile, new data further support the potential of AS-IV as an effective dietary combination for adjuvant therapy of PD.

Objective:To observe the effects of electroacupuncture (EA) on improving motor function and regulating microglial activation based on Notch receptor 1 (Notch1)/Hes family bHLH transcription factor 1 (Hes1) pathway in mice with Parkinson's disease (PD). Methods:Thirty-six male C57BL/6 mice were randomly divided into a control group, a model group and an EA group, 12 mice in each group. PD model was established by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days in the model group and the EA group. From the 1st day of modeling, EA was applied at "Baihui" (GV20) and bilateral "Shenshu" (BL23) in the EA group, with continuous wave, in frequency of 2 Hz and current of 2 mA, 15 min a time, once a day for 14 days continuously. The behavioral performance was evaluated by gait test, pole climbing test and hanging test, the number of positive cells of tyrosine hydroxylase (TH) and the co-expression positive cells of Notch1/ionized calcium binding adaptor molecule 1 (Iba-1) in the substantia nigra of midbrain was assessed by immunofluorescence, the protein expression of TH, α-synuclein (α-syn), Notch1, Hes1, Iba-1, inducible nitric oxide synthase (iNOS), Arginase-1 (ARG1), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and IL-10 was detected by Western blot, the mRNA expression of Notch1 and Hes1 was detected by real-time PCR. Results:Compared with the control group, in the model group, the stride frequency was accelerated (P<0.001) and the stride length was shortened (P<0.001) for the four limbs, the pole climbing test time was prolonged (P<0.01) and the grip level was reduced (P<0.01); in the substantia nigra of midbrain, the number of positive cells of TH was decreased (P<0.001), the number of co-expression positive cells of Notch1/Iba-1 was increased (P<0.001), the protein expression of α-syn, Notch1, Hes1, Iba-1, iNOS, TNF-α, IL-1βand IL-6 was increased (P<0.01, P<0.05, P<0.001), the protein expression of TH, ARG1 and IL-10 was decreased (P<0.01, P<0.001), the mRNA expression of Notch1 and Hes1 was increased (P<0.01). Compared with the model group, in the EA group, the stride frequency was decelerated (P<0.001) and the stride length was increased (P<0.05, P<0.01, P<0.001) for the four limbs, the pole climbing test time was shortened (P<0.05) and the grip level was increased (P<0.05); in the substantia nigra of midbrain, the number of positive cells of TH was increased (P<0.01), the number of co-expression positive cells of Notch1/Iba-1 was decreased (P<0.001), the protein expression of α-syn, Notch1, Hes1, Iba-1, iNOS, TNF-α, IL-6 and IL-1β was decreased (P<0.05, P<0.01), the protein expression of TH, ARG1 and IL-10 was increased (P<0.05, P<0.001, P<0.01), the mRNA expression of Notch1 and Hes1 was decreased (P<0.05). Conclusion:EA can improve the behavioral performance and protect the dopaminergic neurons in PD mice, its mechanism may relate to the inhibition of Notch1/Hes1-mediated neuroinflammation, thus inhibiting the microglial activation.

The present study aimed to investigate the potential role and molecular mechanism of procyanidin B2 (PCB2) in the treatment of multiple sclerosis (MS), with the hope of providing novel strategies for the precision treatment of MS. The target genes of PCB2 and the disease-related target genes of MS were gathered from databases like GEO, GeneCards, OMIM, and DisGeNET. R language was employed for the identification of differentially expressed genes (DEGs), unsupervised clustering analysis, immune infiltration analysis, weighted gene co-expression network analysis (WGCNA), Gene Ontology (GO) analysis, and gene set enrichment analysis (GSEA). Eight machine learning algorithms were employed to screen key genes, and nomograms and ROC curves were constructed to assess the value of the screened biomarker genes in MS diagnosis. Through these analyses, four key genes closely associated with the pathogenesis of MS were selected, namely potassium ion channel subfamily H member 2 (KCNH2), prostaglandin-endoperoxide synthase 1 (PTGS1), estrogen receptor 1 (ESR1), and vascular endothelial growth factor A (VEGFA). These genes were significantly enriched in biological processes related to oxidative stress and served as potential targets for PCB2 in treating MS. Moreover, a key gene-transcription factor (TFs)-microRNA (miRNA) regulatory network was established to preliminarily explore the upstream regulatory mechanisms of these genes. Meanwhile, molecular docking and single-gene GSEA enrichment analysis were carried out to verify the interaction between PCB2 and these key genes. Finally, a CPZ (cuprizone)-induced mouse model of MS was set up, and the effects of PCB2 in MS treatment were verified by means of behavioral tests, pathological staining, immunofluorescence staining, ELISA, RT-PCR, and Western blot detection methods. PCB2 significantly improved behavioral performance in CPZ-induced MS mice, including enhanced motor coordination, reduced anxiety and hyperactivity, and improved spatial learning and memory. Additionally, PCB2 could promote myelin repair and decrease the levels of inflammatory response and oxidative stress in the mouse brain. Specifically, the expression levels of inflammatory factors such as TNF-α, IL-1β, and IL-6 decreased significantly, while the expression level of the anti-inflammatory factor IL-10 increased, and the activities of antioxidant enzymes like CAT, SOD, and GSH-Px increased notably. More importantly, PCB2 could upregulate the expression levels of the KCNH2, PTGS1, ESR1, and VEGFA genes and their related proteins, which may played significant roles in biological processes related to oxidative stress. This study disclosed the potential role of PCB2 in MS treatment by regulating the expression of key genes, providing a theoretical basis and potential therapeutic targets for personalized treatment of MS. PCB2 demonstrates good application prospects in MS treatment. Future research will further validate the clinical application value of these key genes and deeply explore the specific mechanism of PCB2 in MS treatment, aiming to develop more effective treatment regimens for MS.