Epilepsy is a multifactorial neurological disorder with recurrent epileptic seizures. Current research stresses both inflammatory and autoimmune conditions as enablers in the pathophysiological process of epilepsy. In view of the growing concern about the role of adipocytokines in antiepileptic and modulating immune responses, we aimed to investigate the relevance of the adipocytokine signaling pathway in the pathological process of epilepsy and its impacts on peripheral immune characteristics. In this study, expression profiles of 142 peripheral blood samples were downloaded from the Gene Expression Omnibus (GEO) database. Adipocytokine pathway-related genes were screened out by feature selection using machine-learning algorithms. A nomogram was then constructed and estimated for the efficacy of diagnosis. Cluster analysis was employed for the recognization of two distinct epilepsy subtypes, followed by an estimation of the immune cell infiltration levels using single-sample gene-set enrichment analysis (ssGSEA). The biological characteristics were analyzed by functional enrichment analysis. The aberrant regulation of adipocytokine signaling pathway was found in the peripheral blood of patients with epilepsy. Twenty-one differently expressed adipocytokine pathway-related genes were identified and five (RELA, PRKAB1, TNFRSF1A, CAMKK2, and CPT1B) were selected to construct a nomogram. Subsequent validations of its forecasting ability revealed that this model has satisfactory predictive value. The immune cell infiltration degrees, such as those of innate immune cells and lymphocytes, were found to significantly correlate to the levels of adipocytokine pathway-related genes. Additionally, 239 differentially expressed genes (DEGs) were identified and their biological functions were mainly enriched in the regulation of the immune response. In conclusion, our results confirmed the predictive value of adipocytokine pathway-related genes for epilepsy and explored their effects on immune infiltration, thereby improving our understanding of the pathogenesis of epilepsy and providing assistance in the diagnosis and treatment of epilepsy.

Treatment of adipose-derived stem cell (ADSC) substantially improves the neurological deficits during stroke by reducing neuronal injury, limiting proinflammatory immune responses, and promoting neuronal repair, which makes ADSC-based therapy an attractive approach for treating stroke. However, the potential risk of tumorigenicity and low survival rate of the implanted cells limit the clinical use of ADSC. Cell-free extracts from ADSC (ADSC-E) may be a feasible approach that could overcome these limitations. Here, we aim to explore the potential usage of ADSC-E in treating rat transient middle cerebral artery occlusion (tMCAO). We demonstrated that intravenous (IV) injection of ADSC-E remarkably reduces the ischemic lesion and number of apoptotic neurons as compared to other control groups. Although ADSC and ADSC-E treatment results in a similar degree of a long-term clinical beneficial outcome, the dynamics between two ADSC-based therapies are different. While the injection of ADSC leads to a relatively mild but prolonged therapeutic effect, the administration of ADSC-E results in a fast and pronounced clinical improvement which was associated with a unique change in the molecular signature suggesting that potential mechanisms underlying different therapeutic approach may be different. Together these data provide translational evidence for using protein extracts from ADSC for treating stroke.

Mild therapeutic hypothermia has been shown to mitigate cerebral ischemia, reduce cerebral edema, and improve the prognosis of patients with cerebral ischemia. Adipose-derived stem cell-based therapy can decrease neuronal death and infiltration of inflammatory cells, exerting a neuroprotective effect. We hypothesized that the combination of mild therapeutic hypothermia and adipose-derived stem cells would be neuroprotective for treatment of stroke. A rat model of transient middle cerebral artery occlusion was established using the nylon monofilament method. Mild therapeutic hypothermia (33°C) was induced after 2 hours of ischemia. Adipose-derived stem cells were administered through the femoral vein during reperfusion. The severity of neurological dysfunction was measured by a modified Neurological Severity Score Scaling System. The area of the infarct lesion was determined by 2,3,5-triphenyltetrazolium chloride staining. Apoptotic neurons were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. The regeneration of microvessels and changes in the glial scar were detected by immunofluorescence staining. The inflammatory responses after ischemic brain injury were evaluated by in situ staining using markers of inflammatory cells. The expression of inflammatory cytokines was measured by reverse transcription-polymerase chain reaction. Compared with mild therapeutic hypothermia or adipose-derived stem cell treatment alone, their combination substantially improved neurological deficits and decreased infarct size. They synergistically reduced the number of TUNEL-positive cells and glial fibrillary acidic protein expression, increased vascular endothelial growth factor levels, effectively reduced inflammatory cell infiltration and down-regulated the mRNA expression of the proinflammatory cytokines interleukin-1β, tumor necrosis factor-α and interleukin-6. Our findings indicate that combined treatment is a better approach for treating stroke compared with mild therapeutic hypothermia or adipose-derived stem cells alone.

We first assessed the neuroprotective efficacy of mild hypothermia (35°C/24 hours) alone and in combination with magnesium (intravenous loading dose: 360 μmol/kg; 48 hours infusion: 120 μmol/kg/h) commencing 4 hours after global ischemia. Treatment with mild hypothermia alone (CA1 survival rate: 8.7%±0.9%) or with magnesium (9.0%±2.9%) did not significantly increase hippocampal CA1 neuronal survival compared with saline-treated controls (7.1%±0.7%). Next, we assessed mild hypothermia (35°C/24 hours) and moderate hypothermia (33°C/24 hours), alone or in combination with magnesium, when commenced 2 hours after global ischemia. At this time point, all treatments significantly increased CA1 neuronal survival compared with saline controls (CA1 survival rates: mild hypothermia: 13.6%±1.8%; mild hypothermia + magnesium: 19.4%±8.7%; moderate hypothermia: 15.9%±4.1%; moderate hypothermia + magnesium: 21.1%±11.2%; saline control: 6.7%±1.6%; p<0.001). Although a trend for increased neuroprotection was observed when hypothermia was combined with magnesium, it did not reach statistical significance. It also appears that when a 24-hour hypothermia treatment is commenced earlier (≤2 hours postischemia) there is no difference in efficacy between mild and moderate hypothermia.

The tumor microenvironment (TME), with hallmark features of hypoxia and immunosuppression, plays a crucial role in the progression of various solid tumors. However, the intricate interplay between tumor hypoxia and the formation of tumor immune microenvironment in glioma remains incompletely understood. In the present study, we initially identified genes associated with tumor hypoxia and the immune microenvironment through GSEA and IMMPORT database analysis. We subsequently identified hypoxia- and immune-related genes associated with glioma prognosis through further cross-analysis and multidatabase integrated analysis. HSPA5 was ultimately identified as a potential target gene related to the formation of the hypoxic microenvironment and immune microenvironment in glioma. Furthermore, we conducted MTT, colony formation, EdU, migration and invasion assays and intracranial orthotopic tumor model analysis to further evaluate the impact of interfering with HSPA5 expression on the hypoxic and immune microenvironments of glioma. We found that HSPA5 is highly expressed in glioma cells and tissues and is associated with a poor prognosis. Further investigation revealed that hypoxia promotes the malignant biological characteristics of glioma and reshaping the Immunosuppressive phenotype of tumor-associated macrophages (TAMs) through upregulation of the HIF-1α/HSPA5 axis. Silencing HSPA5 alleviated glioma hypoxia tolerance and induced the polarization of TAMs toward the M1 phenotype. The induced macrophages could exhibit a tumor-suppressive effect. These observations suggest that HSPA5 upregulation promotes glioma progression by inducing hypoxia tolerance and reshaping the Immunosuppressive phenotype of TAMs. Therefore, targeting HSPA5 may be a novel therapeutic strategy for glioma.