Background and Purpose: Neuroinflammation has been shown to play a critical role in secondary craniocerebral injury, leading to poor outcomes for TBI patients. Abrocitinib, a Janus kinase1 (JAK1) selective inhibitor approved to treat atopic dermatitis (AD) by the Food and Drug Administration (FDA), possesses a novel anti-inflammatory effect. In this study, we investigated whether abrocitinib could ameliorate neuroinflammation and exert a neuroprotective effect in traumatic brain injury (TBI) models. Methods: First, next-generation sequencing (NGS) was used to select genes closely related to neuroinflammation after TBI. Then, magnetic resonance imaging (MRI) was used to dynamically observe the changes in traumatic focus on the 1st, 3rd, and 7th days after the induction of fluid percussion injury (FPI). Moreover, abrocitinib’s effects on neurobehaviors were evaluated. A routine peripheral blood test was carried out and Evans blue dye extravasation, cerebral cortical blood flow, the levels of inflammatory cytokines, and changes in the numbers of inflammatory cells were evaluated to investigate the function of abrocitinib on the 1st day post-injury. Furthermore, the JAK1/signal transducer and activator of transcription1 (STAT1)/nuclear factor kappa (NF-κB) pathway was assessed. Results: In vivo, abrocitinib treatment was found to shrink the trauma lesions. Compared to the TBI group, the abrocitinib treatment group showed better neurological function, less blood-brain barrier (BBB) leakage, improved intracranial blood flow, relieved inflammatory cell infiltration, and reduced levels of inflammatory cytokines. In vitro, abrocitinib treatment was shown to reduce the pro-inflammatory M1 microglia phenotype and shift microglial polarization toward the anti-inflammatory M2 phenotype. The WB and IHC results showed that abrocitinib played a neuroprotective role by restraining JAK1/STAT1/NF-κB levels after TBI. Conclusions: Collectively, abrocitinib treatment after TBI is accompanied by improvements in neurological function consistent with radiological, histopathological, and biochemical changes. Therefore, abrocitinib can indeed reduce excessive neuroinflammation by restraining the JAK1/STAT1/NF-κB pathway.

Aims: Annexin A5 (ANXA5) exhibited potent antithrombotic, antiapoptotic, and anti-inflammatory properties in a previous study. The role of ANXA5 in traumatic brain injury (TBI)-induced intestinal injury is not fully known. Main methods: Recombinant human ANXA5 (50 µg/kg) or vehicle (PBS) was administered to mice via the tail vein 30 min after TBI. Mouse intestine tissue was gathered for hematoxylin and eosin staining 0.5 d, 1 d, 2 d, and 7 d after modeling. Intestinal Western blotting, immunofluorescence, TdT-mediated dUTP nick-end labeling staining, and enzyme-linked immunosorbent assays were performed 2 days after TBI. A series of kits were used to assess lipid peroxide indicators such as malonaldehyde, superoxide dismutase activity, and catalase activity. Key findings: ANXA5 treatment improved the TBI-induced intestinal mucosa injury at different timepoints and significantly increased the body weight. It significantly reduced apoptosis and matrix metalloproteinase-9 and inhibited the degradation of tight-junction-associated protein in the small intestine. ANXA5 treatment improved intestinal inflammation by regulating inflammation-associated factors. It also mitigated the lipid peroxidation products 4-HNE, 8-OHDG, and malonaldehyde, and enhanced the activity of the antioxidant enzymes, superoxide dismutase and catalase. Lastly, ANXA5 significantly enhanced nuclear factor E2-related factor 2 (Nrf2) and hemeoxygenase-1, and decreased high mobility group box 1 (HMGB1). Significance: Collectively, the results suggest that ANXA5 inhibits TBI-induced intestinal injury by restraining oxidative stress and inflammatory responses. The mechanisms involved sparking the Nrf2/hemeoxygenase-1-induced antioxidant system and suppressing the HMGB1 pathway. ANXA5 may be an attractive therapeutic candidate for protecting against TBI-induced intestinal injury.

Brain induced extracellular vesicle (BDEV) elevates after traumatic brain injury (TBI) and contributes to secondary brain injury. However, the role of BDEV in TBI remains unclear. In this study, we determined the mechanisms of BDEV in brain injury and explored whether neuroprotective drug BKca channel opener NS1619 may attenuate BDEV‐induced brain injury. We injected BDEV and lactadherin, respectively, to mimic the up and downregulation of BDEV after TBI and illustrated the role of BDEV in vivo. In vitro, the membrane potential and calcium concentration of HT‐22, bEnd3, and BV‐2 were measured by fluorescent staining. The effects of BDEV and NS1619 on HT‐22 were evaluated by CCK‐8, LDH release assay, Na⁺/k⁺‐ATPase activity, JC‐1 staining, DHE staining, and 4‐HNE staining, respectively. The role of BDEV and NS1619 on the Nrf2/HO‐1/p65 pathway was also evaluated in HT‐22. Finally, we administrated TBI mice with NS1619 to clarify the role of NS1619 against BDEV in vivo. Our results suggested that BDEV aggravated and lactadherin mitigated TBI‐induced EB leakage, brain edema, neuronal degeneration, apoptosis, ROS level, microgliosis, MMP‐9 activity, and NF‐κB activation. In vitro, BDEV‐caused depolarized membrane potential and calcium overload were significantly attenuated by NS1619 in HT‐22, bEnd3, and BV‐2. BDEV markedly decreased cell viability, Na⁺/k⁺‐ATPase activity, and caused mitochondrial dysregulation, oxidative stress, and NF‐ĸB activation. NS1619 pretreatment alleviated above process and enhanced antioxidant system Nrf2/HO‐1 in HT‐22. Finally, NS1619 administration significantly inhibited neuroinflammation response and improved TBI outcome after TBI. NS1619 treatment also reduced 4‐HNE content and NF‐ĸB activation and enhanced Nrf2/HO‐1 pathway. Our data showed that BDEV aggravated brain injury by perturbing cell membrane potential, calcium homeostasis, oxidative stress, and neuroinflammation. The BKca channel opener NS1619 attenuated BDEV‐induced pathological process in vitro and in vivo by modulating the BKca channel and Nrf2/HO‐1/NF‐ĸB pathway.

Background: Patients with cancer often develop a prothrombotic state that can evolve into venous and arterial thrombosis, which is associated with poor clinical outcomes. Glioblastoma multiforme (GBM) is the most frequently associated with thrombosis, but the underlying causes of this prothrombotic state are poorly defined. Objectives: We designed a study to characterize the expression of coagulation factor X (FX) and its alternatively spliced transcripts in glioblastoma tissues surgically removed from patients and in clonal cell lines. Methods: The F10 mRNA and FX protein were quantified in tissues surgically removed from seven patients with glioblastoma (glioma grade 3–4) and those from non-tumor patients. Glioblastoma cells from three human clonal lines were examined for the expression and secretion of FX at baseline and after the cells were stimulated with lipopolysaccharide (LPS) or subjected to oxygen/glucose starvation in culture. PCR products were subjected to Sanger sequencing and amplicon sequencing to identify F10 isoforms and their ratios. A chromogenic assay was performed to assess FX activity. Results: Glioblastoma tissue and cell lines expressed high levels of the full-length and an alternatively spliced F10 mRNA. The latter produced a C-terminal truncated FX. The ratio of full-length to truncated F10 transcripts was significantly higher in normal brain tissues than in glioblastoma tissue. In cultured cells from the glioblastoma cell lines, FX was secreted to the conditioned medium and was active in cleaving a chemical substrate. The FX expression and secretion were upregulated in cells stimulated with LPS or subjected to oxygen/glucose starvation. Discussion: Glioblastoma cells synthesize and secrete FX that was active in promoting thrombin generation. These findings provide a new underlying mechanism to explain why glioblastoma patients are prone to developing thrombosis.