BV2 (小鼠小胶质细胞)（STR鉴定）

Normal tension glaucoma (NTG) is a predominant subset of glaucoma in Asia and is characterized by glaucomatous optic neuropathy in the absence of elevated intraocular pressure. Alterations in retinal blood vessels are reported to be important mechanisms of glaucomatous optic nerve damage. Retinal peripapillary vascular density is assessed in patients with early stage NTG and OPTN (E50K) mutant mice and confirmed a similar reduction in retinal peripapillary vascular density in patients with NTG and model mice. Thus, identifying the mechanisms underlying retinal vascular changes in NTG is crucial for developing effective therapeutic strategies. This study revealed that Rpl17 is upregulated in the retinal microglia of OPTN (E50K) mutant mice. Rpl17 exerts regulatory control over Apoa1 by directly interacting with Stat5b, which causes damage to retinal vascular endothelial cells and leads to a reduction in retinal peripapillary vascular density. Additionally, we identified ellagic acid (EA) as an Apoa1 antagonist that is able to alleviate damage to retinal vascular endothelial cells and increase retinal peripapillary vascular density, which subsequently protected retinal ganglion cells and improved visual function. The work elucidates the vascular mechanism of NTG optic nerve damage and proposes EA as an effective adjuvant therapy strategy.

Aging is associated with increased retinal cell apoptosis, which contributes to decreases in retinal function. Apoptotic retinal cell clearance relies on microglial efferocytosis, but the impact of aging on this process has not been fully elucidated. In this study, we aimed to shed light on this by using single-cell RNA sequencing (sc-RNA-seq) to compare young and aged mouse retinal transcriptional profiles, in which 74,412 retinal cells from young and aged mice were classified into 10 transcriptionally distinct retinal cell types, and differentially expressed genes between young versus aged retinas were mainly associated with cellular senescence and apoptosis. Furthermore, ligand–receptor interactions (e.g., AXL-GAS6, MERTK-GAS6) between microglia and other retinal cells were strengthened in aged, compared to young retinas. Additionally, among microglia, Subcluster 4 was found under partial clustering to be associated with efferocytosis, of which aged microglia had downregulated efferocytosis-associated genes. The impact of aging on microglial efferocytosis was further verified in vitro by doxorubicin (DOX)-induced senescent BV2 microglia, and in vivo by a retinal ischemia/reperfusion (I/R) injury mouse model. In vitro, DOX-treated BV2 microglia had significantly lowered efferocytosis, as well as efferocytosis-related MerTK and Axl protein expression; this was also present in vivo in aged retinas post-I/R injury, with increased co-localization of ionized calcium-binding adapter molecule 1 + microglia with apoptotic retinal cells, along with reduced efferocytosis-related protein expression. Overall, microglial efferocytosis of apoptotic cells decreased with aging, suggesting that modulating this process could serve as a possible therapeutic target for age-related retinal diseases.

In recent years, the application of engineered NMts has significantly contributed to various biomedical fields. ZnO NMts (ZnO NMts) are widely utilized due to their biocompatibility, unique physical and chemical properties, stability, and cost-effectiveness for large-scale production. They have emerged as potential materials for anti-cancer applications. This study aims to study the impact of ZnO Nanorod flowers (ZnO NRfs) and their combination with temozolomide (TMZ) on glioma cells. Normal mouse microglia (BV2) will be used as a control to assess the effects on mouse glioma cells (G422) and human glioma cells (LN229). The effects of these substances were evaluated on G422 and LN229 cells through various parameters such as IC50 value, Zn2+ accumulation, ROS production, apoptosis, mitochondrial membrane potential (MMP) depolarization, and examination of organelles like mitochondria and lysosomes. Additionally, hypoxia-inducible factor-1α (HIF-1α), endothelial cell PAS domain protein 1 (EPAS1), autophagy markers (LC3), mitophagy and phagocytosis marker (BNIP3) were assessed. The results demonstrated that the combination of ZnO NRfs and TMZ could influence the expression of HIF-1α, EPAS1, LC3, and BNIP3 proteins, leading to mitophagy in glioma cells. This combination treatment has the potential to effectively eliminate glioma cells by activating the mitophagy pathway, which provides a good prospect for the clinical treatment of glioma.

Introduction: Glaucoma is a prevalent cause of irreversible vision impairment, characterized by progressive retinal ganglion cells (RGCs) loss, with no currently available effective treatment. Rapamycin (RAPA), an autophagy inducer, has been reported to treat glaucoma in rodent models by promoting RGC survival, but its limited water solubility, systemic toxicity, and pre-treatment requirements hinder its potential clinical applications.Methods: Chitosan (CS)-RAPA carbon dot (CRCD) was synthesized via hydrothermal carbonization of CS and RAPA and characterized by transmission electron microscopy, Fourier transform infrared spectra, and proton nuclear magnetic resonance. In vitro assays on human umbilical cord vein endothelial and rat retinal cell line examined its biocompatibility and anti-oxidative capabilities, while lipopolysaccharide-stimulated murine microglia (BV2) assays measured its effects on microglial polarization. In vivo, using a mouse retinal ischemia/reperfusion (I/R) model by acute intraocular pressure elevation, the effects of CRCD on visual function, RGC apoptosis, oxidative stress, and M2 microglial polarization were examined.Results: CRCD exhibited good water solubility and anti-oxidative capabilities, in the form of free radical scavenging. In vitro, CRCD was bio-compatible and lowered oxidative stress, which was also found in vivo in the retinal I/R model. Additionally, both in vitro with lipopolysaccharide-stimulated BV2 cells and in vivo with the I/R model, CRCD was able to promote M2 microglial polarization by activating autophagy, which, in turn, down-regulated pro-inflammatory cytokines, such as IL-1β and TNF-α, as well as up-regulated anti-inflammatory cytokines, such as IL-4 and TGF-β. All these anti-oxidative and anti-inflammatory effects ultimately aided in preserving RGCs, and subsequently, improved visual function.Discussion: CRCD could serve as a potential novel treatment strategy for glaucoma, via incorporating RAPA into CDs, in turn not only mitigating its toxic side effects but also enhancing its therapeutic efficacy.

Microbiota dysfunction induces intestinal disorders and neurological diseases. Mannuronate oligosaccharides (MAOS), a kind of alginate oligosaccharide (AOS), specifically exert efficacy in shaping gut microbiota and relieving cognitive impairment. However, the key regulatory factors involved, such as the specific strains and metabolites as well as their regulatory mechanisms, remain unclear at present. This research investigates how MAOS specifically impact the gut–brain axis in vivo and in vitro. The results showed that pretreatment with MAOS significantly ameliorated dextran sodium sulfate (DSS)-induced colitis and secondary nerve injury. This preventive mechanism operates through the regulation of serum DOPC abundance and the gut–brain axis, achieved by inhibiting the TLR4/MyD88/NF-κB pathway. These findings underscore the crucial role of dietary MAOS in the prevention of colitis and neurological disorders, providing a rationale for the application of MAOS in disease prevention and functional food ingredients.

Normal-tension glaucoma (NTG) is a subtype of primary open-angle glaucoma (POAG). Patients with NTG still experience significant optic nerve damage despite maintaining normal intraocular pressures. The mechanism of optic nerve damage in glaucoma with normal pressure is still unclear. Research has shown that OPTN (E50K) mutations exacerbate the inflammatory response of retinal microglia. However, there is still a lack of evidence on how OPTN (E50K) mutations directly regulate their inflammatory pathways through key molecules. This study explores the role of microglial inflammation caused by the interaction between IRF7 and NLRP3 molecules in NTG optic nerve injury. Single-cell RNA sequencing (scRNA-seq) was employed to analyze retinal microglial cells from both wild-type (WT) and OPTN (E50K) mutant mice. The analysis revealed significant enrichment of inflammatory pathways and a critical role of IRF7 in modulating NLRP3 activation. Techniques such as Western blot (WB), qPCR, immunofluorescence (IF), and molecular docking were utilized to confirm the interactions between IRF7 and NLRP3. The findings demonstrate that the OPTN (E50K) mutation reduces the suppressive effect of IRF7 on NLRP3, leading to a pro-inflammatory microglial phenotype and exacerbating the optic nerve damage of NTG. This study provides a new therapeutic target for the treatment of NTG optic nerve damage.

Background: The increasing aged population poses issues in the management of age-related disorders, notably Alzheimer’s disease (AD), which significantly affects the health and quality of life of seniors. Neuroinflammation is a significant factor in Alzheimer’s disease pathogenesis. Isoliensinine (ISO), a bisbenzylisoquinoline alkaloid derived from lotus seed embryos, exhibits antioxidant and anti-inflammatory effects. Nonetheless, its function in neuroinflammation has yet to be investigated. Methods: We examined the impact of ISO on LPS-induced neuroinflammation in BV2 microglial cells by using biological tests. Western blotting confirmed ISO’s influence on MAPK/NF-κB signaling pathways. In addition, oxidative stress markers and JC-1 staining were employed to assess the impact of ISO on LPS-induced oxidative stress and mitochondrial dysfunction in BV2 cells. Results: ISO markedly diminished LPS-induced neuroinflammation in BV2 cells through the modulation of the MAPK/NF-κB pathway. Conditioned media derived from ISO-treated BV2 cells enhanced the vitality of HT-22 cells. ISO also alleviated oxidative stress and mitochondrial dysfunction. Conclusion: Our findings indicate that ISO mitigates neuroinflammation by inhibiting MAPK/NF-κB signaling and provides neuroprotection by diminishing oxidative stress and mitochondrial impairment. These effects collectively enhance its neuroprotective capacity, indicating that ISO may represent a potential candidate for further investigation in AD.

This study aims to investigate the effect of exosomes derived from olfactory mucosa mesenchymal stem cells (OM-MSCs-Exo) on microglial polarization and its potential therapeutic role in Alzheimer’s disease (AD). OM-MSCs-Exo were isolated and purified from the mice olfactory mucosa, followed by phenotypic characterization. Proteins transferred by OM-MSCs-Exo were screened using proteomic analysis. The AD model was established in microglial cells and mice with Aβ 1–42 . Immunofluorescence and biochemical assays were employed to assess the impact of OM-MSCs-Exo and its secreted protein FGFR1 on microglial polarization. Protein–protein interactions and immunoprecipitation were used to identify the target proteins of FGFR1 in microglial cells. Additionally, the effects of OM-MSCs-Exo-induced microglial polarization on neuronal inflammation and cognitive function in mice were evaluated. OM-MSCs-Exo were successfully isolated and purified. FGFR1 was significantly upregulated in OM-MSCs-Exo compared to OM-MSCs. Aβ 1–42 induced M1 polarization and suppressed M2 polarization of microglia, which was reversed by OM-MSCs-Exo. FGFR1 overexpression in OM-MSCs-Exo further enhanced M2 polarization in microglial cells. Phospholipase C gamma 1 (PLCγ1) was identified as the target of FGFR1, and knocking down PLCγ1 reversed the effects of FGFR1-overexpressing OM-MSCs-Exo. OM-MSCs-Exo alleviated cognitive decline and neuroinflammation in AD mice, with FGFR1 overexpression further enhancing these effects. OM-MSCs-Exo promote M2 polarization of microglia in AD mice through the FGFR1/PLCγ1 pathway, alleviating neuronal inflammation and cognitive dysfunction.

Background Microglia-mediated neuroinflammation is closely related to the development of Alzheimer's disease (AD). This study further elucidated the regulatory mechanism of microglia polarization in AD. Method Microglia polarization was assessed using RT-qPCR, ELISA, and immunofluorescence (IF). Western blot (WB) analyzed inflammation-related, p-tau, and apoptosis-related proteins. Neuronal damage was evaluated by immunofluorescence, and neuronal apoptosis by flow cytometry and TUNEL assay. METTL3 and IκBα expression were detected using RT-qPCR and WB. N 6 -methyladenosine (m 6 A) levels were quantified with a colorimetric assay. RNA pull-down assay examined METTL3, IGF2BP2, and IκBα mRNA binding. IGF2BP expression was assessed by RT-qPCR. Learning and memory abilities were evaluated using morris water maze (MWM) test and novel object recognition (NOR) test. Inflammation-related proteins were detected using IF. Results Stimulation with Aβ 1-42 led to microglia M1 polarization, upregulation of inflammation-related proteins, and exacerbation of neuronal injury and apoptosis, along with increased p-tau expression in neurons. METTL3/IGF2BP2 modulated IκBα m 6 A modification through binding to IκBα mRNA, enhancing its expression. Enhanced METTL3 or IGF2BP2 expression suppressed M1 polarization, inflammation, and neuronal apoptosis in microglia, reversed by knockdown of IκBα. AD model mice exhibited cognitive impairments, neuroinflammation, and elevated M1 polarization. METTL3 or IGF2BP2 overexpression improved cognitive function, reduced neuroinflammation, and inhibited M1 polarization, and this effect was similarly reversed by knockdown of IκBα. Conclusion Our study demonstrates that the METTL3/IGF2BP2/IκBα axis is involved in neuroinflammation in AD by modulating microglia M1/M2 polarization, which sheds light on the treatment of AD.

Microglia and exosomes are intimately connected with the pathogenesis of Alzheimer’s disease (AD). We aim to investigate the role and potential mechanisms of M2-like (anti-inflammatory) microglia-derived exosomes (M2-Exos) in AD. We utilized an Aβ 1−42 -induced AD model in HT-22 neurons and mouse. The effects of M2-Exo on mitochondrial damage, ferroptosis, oxidative stress, and inflammation levels in the AD cell/animal models were evaluated using transmission electron microscopy, immunoblotting, and biochemical assay kits. Cognitive function in mouse was assessed through behavioral tests. In the AD cell/animal models, the effects of M2-Exo on the Wnt/β-catenin pathway were investigated through immunofluorescence and immunoblotting. AD cells were treated with HLY78 (Wnt/β-catenin pathway activator) to explore the modulation of the pathway. After knocking down TREM2 in M2-Exo, mitochondrial damage, ferroptosis, oxidative stress, and inflammation markers were reevaluated in AD cell and animal models. Aβ 1−42 induced mitochondrial shrinkage and deformation in neurons, upregulated ACSL4, PTGS2, Fe2+/Fe, lipid peroxide (LPO), ROS, MDA, IL-6, IL-1β, and TNF-α, while it downregulated GPX4, FTH1, and GSH-PX. M2-Exo reversed the effects induced by Aβ 1−42 both in vitro and in vivo, and M2-Exo improved cognitive function in AD mouse. HLY78 also reversed the effects induced by Aβ 1−42 . M2-Exo increased the levels of β-catenin. BV2 cells converting to M2-like type increased TREM2 levels. Knocking down TREM2 in M2-Exos resulted in decreased neuronal β-catenin levels, reversing the beneficial effects of M2-Exo on AD cell and mouse models. M2-Exo TREM2 alleviates neuronal ferroptosis, inflammation, and oxidative stress in AD by activating the Wnt/β-catenin signaling pathway.