Neuroinflammatory responses have been recognized as an important aspect in the pathogenesis of Parkinson’s disease (PD). Transcriptional regulation plays a critical role in the process of inflammation. Transcription factor myocyte enhancer factor 2D (MEF2D) is identified as a central factor in transmission of extracellular signals and activation of the genetic programs in response to a wide range of stimuli in several cell types, including neurons. But its presence and function in microglia have not been reported. We therefore investigated the effect of MEF2D in activated microglia on the progress of neuroinflammation and the survival of neurons. BV2 cells and primary cultured glial cells were stimulated with lipopolysaccharide (LPS). Samples from cells were examined for MEF2D expression, interleukin-10 (IL-10), and tumor necrosis factor alpha (TNF-α) by immunoblotting, quantitative real-time PCR (qPCR) or enzyme-linked immunosorbent assay (ELISA). The activity of MEF2D was examined by electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP). Recombinant lentivirus expressing shRNA specific to MEF2D was used to silence MEF2D expression in BV2 cells. The role of IL-10 transcriptionally induced by MEF2D on neuronal survival was assessed by anti-IL-10 neutralizing antibody. The survival of neurons was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Male C57bl/6 mice were used to establish an acute PD model. Brain sections and cell slides were tested by immunofluorescence. We demonstrated that MEF2D was present in microglia. Activation of microglia was associated with an increase in MEF2D level and activity in response to different stimuli in vivo and in vitro. MEF2D bound to a MEF2 consensus site in the promoter region of IL-10 gene and stimulated IL-10 transcription. Silencing MEF2D decreased the level of IL-10, increased the TNF-α mRNA, and promoted inflammation-induced cytotoxicity, consistent with the result of inhibiting IL-10 activity with an anti-IL-10 neutralizing antibody. Our study identifies MEF2D as a critical regulator of IL-10 gene expression that negatively controls microglia inflammation response and prevents inflammation-mediated cytotoxicity.

An abstract is unavailable. This article is available as a PDF only.

Oxidative stress and mitochondrial dysfunction play critical roles in ischemia/reperfusion (I/R) injury. DJ-1 is an endogenous antioxidant that attenuates oxidative stress and maintains mitochondrial function, likely acting as a protector of I/R injury. In the present study, we explored the protective effect of a possible DJ-1 agonist, sodium phenylbutyrate (SPB), against I/R injury by protecting mitochondrial dysfunction via the up-regulation of DJ-1 protein. Pretreatment with SPB up-regulated the DJ-1 protein level and rescued the I/R injury-induced DJ-1 decrease about 50 % both in vivo and in vitro. SPB also improved cellular viability and mitochondrial function and alleviated neuronal apoptosis both in cell and animal models; these effects of SPB were abolished by DJ-1 knock-down with siRNA. Furthermore, SPB improved the survival rate about 20 % and neurological functions, as well as reduced about 50 % of the infarct volume and brain edema, of MCAO mice 23 h after reperfusion. Therefore, our findings demonstrate that preconditioning of SPB possesses a neuroprotective effect against cerebral I/R injury by protecting mitochondrial function dependent on the DJ-1 up-regulation, suggesting that DJ-1 is a potential therapeutic target for clinical ischemic stroke.

Cerebral hemodynamic transformation is a relatively common finding in patients with traumatic brain injury (TBI). Knowledge of cerebral hemodynamic disturbance may assist in predicting the management outcome. Transcranial Doppler ultrasonography (TCD) monitoring of patients with TBI can be used to reveal various pathologic hemodynamic changes. The objective of this study was to compare the clinical outcomes of postoperative routine intracranial pressure (ICP) monitoring versus ICP monitoring combined with TCD monitoring in patients with brain trauma after decompressive craniectomy. This was a retrospective study of 30 patients with TBI who underwent ICP combined with TCD monitoring (after 2015) compared with a historical control group of 30 patients who only underwent routine ICP monitoring (in 2013-2014). ICP, partial pressure of carbon dioxide, hemoglobin, and hematocrit values were monitored and recorded on a daily basis for 7 days after operation. Neuroimaging was also performed at admission. Neurologic outcome was assessed at 2 weeks and 6 months after operation using the Glasgow Outcome Score Extended (GOS-E). Unconditional multivariable logistic regression was conducted to analyze the factors for favorable clinical outcome. Two weeks after operation, there were no differences in mortality rate between the 2 groups (P = 0.643). When considering the GOS-E score at 6 months, there were no differences in clinical prognosis between the 2 groups (P = 0.101), but the ICP combined with TCD monitoring group showed a higher frequency of patients with favorable outcome compared with the routine ICP monitoring group (P = 0.043). Unconditional multivariable logistic regression results showed that no factor was independently associated with GOS-E at 6 months. TCD could be helpful for the serial monitoring of cerebral hemodynamic changes after decompressive craniectomy for TBI, which could be beneficial for neurologic outcome improvement.

Background: Calcium signaling is implicated in multiple processes including immune response that important in tumor progression. Kidney renal clear cell carcinoma (KIRC) is the most frequent histological type of renal cell carcinoma with up to a third of cases develop metastases. As a result of a lack of in-depth understanding of the mechanisms underlying KIRC, treatment options have been limited. Here, we aim to comprehensively investigate the landscape of Ca²⁺ channels, pumps and exchangers in KIRC patients.Methods: The mRNA expression profiles and gene variations of 58 calcium-related genes (CRGs) in KIRC patients and normal control cases were downloaded from TCGA database. CRGs-related risk score was constructed to quantify calcium patterns by using least absolute shrinkage and selection operator (LASSO) regression. The prognostic value, biological functions, immune landscape and therapeutic sensitivities based on CRGs-related risk score were then evaluated using multiple methods. Finally, key gene of CRGs was identified by weighted gene co-expression network analysis (WGCNA). TCGA-CPTAC, GSE53757 datasets, as well as human tissues were used for validation.Results: KIRC patients had significant differences in CRG expression, prognosis, and biological functions between two CRG clusters. CRGs-related risk score was then determined. The prognosis, tumor mutation burden, immune cell infiltration, immune checkpoints, and the response of targeted inhibitors were remarkably different between high and low CRGs-related risk subtypes. CRGs-related high-risk subtype was characterized by immunosuppressive microenvironment with poor prognosis. Meanwhile, several targeted drugs showed distinct sensitivity between CRGs-related risk subtypes. Finally, TRPM3 was identified as a key CRG based on risk score in KIRC patients. TRPM3 mRNA and protein expression were significantly lower in KIRC tumors than in normal controls. Low TRPM3 expression was associated with poor prognosis in KIRC patients.Conclusion: Our study highlighted the promising prognostic value of CRGs in KIRC tumors. The evaluation of CRGs-related risk score will contribute to predicting prognosis and clinical therapy in KIRC patients.