N-n-butyl haloperidol iodide mediates cardioprotection via regulating AMPK/FoxO1 signalling.

Derangement of redox condition largely contributes to cardiac ischemia/reperfusion (I/R) injury. FoxO1 is a transcription factor which transcripts a series of antioxidants to antagonize I/R-induced oxidative myocardial damage. N-n-butyl haloperidol iodide (F2 ) is a derivative derived from haloperidol structural modification with potent capacity of inhibiting oxidative stress. This investigation intends to validate whether cardio-protection of F2 is dependent on FoxO1 using an in vivo mouse I/R model and if so, to further elucidate the molecular regulating mechanism. This study initially revealed that F2 preconditioning led to a profound reduction in I/R injury, which was accompanied by attenuated oxidative stress and upregulation of antioxidants (SOD2 and catalase), nuclear FoxO1 and phosphorylation of AMPK. Furthermore, inactivation of FoxO1 with AS1842856 abolished the cardio-protective effect of F2 . Importantly, we identified F2 -mediated nuclear accumulation of FoxO1 is dependent on AMPK, as blockage of AMPK with compound C induced nuclear exit of FoxO1. Collectively, our data uncover that F2 pretreatment exerts significant protection against post ischemic myocardial injury by its regulation of AMPK/FoxO1 pathway, which may provide a new avenue for treating ischemic disease.

Serum uric acid predicts the development of atherosclerosis in women but not in men: A ten-year cohort study in China.

BACKGROUND AND AIM: Atherosclerosis is becoming a significant health burden. Serum uric acid (SUA) is the final enzymatic product of purine metabolism and can contribute to the development of atherosclerosis. The aim of this study was to explore the possible predictive value of SUA in the development of atherosclerosis in a healthy Chinese population. METHODS AND RESULTS: In this study, a total of 11,222 healthy subjects with no carotid plaque at baseline were enrolled and divided into sex-specific groups, and then the occurrence of carotid plaque during the follow-up time was documented. The association between carotid plaque and SUA levels was examined using Cox proportional-hazards regression models. The mean SUA level was 5.35 ± 1.41 mg/dL. A total of 2,911 individuals (25.94%) developed carotid plaque during the follow-up time, including 1,071 females and 1,840 males. After adjusting for potential confounding factors, the hazard ratio (HR) and 95% confidence interval (95% CI) in women for the occurrence of carotid plaque associated with SUA levels were 1.163 (1.017-1.330), but no significant correlation was found in men, as the HR was 1.050 (0.965-1.143). CONCLUSION: Our results indicate that SUA levels predict the development of carotid plaque independent of traditional risk factors only in women.

Taurine promotes axonal sprouting via Shh-mediated mitochondrial improvement in stroke.

PURPOSE: Motor function is restored by axonal sprouting in ischemic stroke. Mitochondria play a crucial role in axonal sprouting. Taurine (TAU) is known to protect the brain against experimental stroke, but its role in axonal sprouting and the underlying mechanism are unclear. METHODS: We evaluated the motor function of stroke mice using the rotarod test on days 7, 14, and 28. Immunocytochemistry with biotinylated dextran amine was used to detect axonal sprouting. We observed neurite outgrowth and cell apoptosis in cortical neurons under oxygen and glucose deprivation (OGD), respectively. Furthermore, we evaluated the mitochondrial function, adenosine triphosphate (ATP), mitochondrial DNA (mtDNA), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PCG-1α), transcription factor A of mitochondria (TFAM), protein patched homolog 1 (PTCH1), and cellular myelocytomatosis oncogene (c-Myc). RESULTS: TAU recovered the motor function and promoted axonal sprouting in ischemic mice. TAU restored the neuritogenesis ability of cortical neurons and reduced OGD-induced cell apoptosis. TAU also reduced reactive oxygen species, stabilized mitochondrial membrane potential, enhanced ATP and mtDNA content, increased the levels of PGC-1α, and TFAM, and restored the impaired levels of PTCH1, and c-Myc. Furthermore, these TAU-related effects could be blocked using an Shh inhibitor (cyclopamine). CONCLUSION: Taurine promoted axonal sprouting via Shh-mediated mitochondrial improvement in ischemic stroke.

Longitudinal Associations Between Serum Bilirubin Level and Carotid Atherosclerosis Plaque in a Health Screening Population.

This study aimed to determine the relationship between bilirubin levels and carotid atherosclerosis (CAS) in the health screening population. After propensity score matching, this retrospective cohort study included 4360 subjects who underwent health examinations regularly in Hebei General Hospital between January 2010 and December 2019 and had no carotid plaque at baseline. After an average follow-up of 26.76 months, the main endpoint Cox regression analysis of carotid plaques was performed. After adjusting the confounding factors, Cox regression analysis showed that when serum total bilirubin (TBIL) and unconjugated bilirubin (UCB) increased by 1 standard deviation (SD), the risk of carotid plaque decreased by 7.30% (95% confidence interval (CI): 2.80-11.60%) and 15.70% (95% CI: 11.40-19.80%), respectively. When conjugated bilirubin (CB) increased by 1 SD, the risk of carotid plaques increased by 24.3% (95% CI: 19.7-29.0%). TBIL and UCB levels were negatively associated with CAS, and CB levels were positively associated with CAS.

The construction of neurogenesis-related ceRNA network of ischemic stroke treated by oxymatrine.

BACKGROUND: Known as a disease associated with high mortality, disability and a significant financial burden, ischemic stroke ranks as one of the three diseases threatening human health. Recent advances in omics technology created opportunities to uncover the mechanism in ischemic stroke occurrence and treatment. In this study, we aimed to construct the competitive endogenous RNA (ceRNA) networks of ischemic stroke treated by oxymatrine intervention. METHOD: The middle cerebral artery occlusion (MCAO) mouse model of ischemic stroke was constructed, and oxymatrine was administered. Then RNA-Sequencing was performed and integrated analysis of mRNAs, lncRNAs and circRNAs was conducted to reveal the pharmacology of oxymatrine. Functional enrichment analysis was performed to explore the underlying mechanism of differentially expressed (DE) mRNAs. The protein-protein interaction (PPI) network of neurogenesis-related genes and long noncoding RNAs (lncRNAs)/circular RNAs (circRNAs) based ceRNA networks were constructed. RESULTS: First, this study revealed the DE-mRNAs, DE-lncRNAs and DE-circRNAs between Oxymatrine treated group and the MCAO group. Then, the common 1231 DE-mRNAs, 32 DE-lncRNAs and 31 DE-circRNAs with opposite trends were identified. The Kyoto Encyclopedia of Genes and Genomes to identify the functional enrichment of 1231 DE-mRNAs were enriched in neurogenesis-related biological processes. Based on neurogenesis-related DE-mRNAs, the PPI network was constructed, and hub genes were identified based on centrality. Finally, both the lncRNA-based and circRNAs-based ceRNA networks were constructed. CONCLUSION: In summary, this study identified novel coding and noncoding ischemic stroke targets of oxymatrine-treated MCAO. Most importantly, we identified lncRNAs and circRNAs candidates as potential oxymatrine targets and constructed the neurogenesis-related ceRNA networks.

The Epigenetic Regulation of OLIG2 by Histone Demethylase KDM6B in Glioma Cells.

Gliomas are common tumors that occur in the brain, accounting for 80% of all malignant brain tumors. Oligodendrocyte transcription factor 2 (OLIG2) is a key transcription factor and strongly expressed in gliomas, which drives proliferation and invasion of glioma cells. Our previous studies have shown that histone lysine (K) demethylase 6B (KDM6B) promotes glioma development. The data also showed that OLIG2 content was positively correlated with KDM6B. Based on this, we proposed that KDM6B may play biological roles by regulating OLIG2 expression. Subsequently, many experiments were performed including specific inhibitor treatment, gene knockdown, and chromatin immunoprecipitation (ChIP) array. These results indicated that inhibition of KDM6B enzymatic activity with GSK-J4 reduces OLIG2 gene expression and protein content. The KDM6B knockdown experiment yielded similar results, that is, it reduces the mRNA and protein level of OLIG2 in glioma cells. ChIP assay showed that the promoter of OLIG2 can be bound by KDM6B, which catalyzes the demethylation of H3K27me3 and increases the expression of OLIG2. This study reveals a new regulatory mechanism of OLIG2 by KDM6B, which has important implications for the future development of drugs for gliomas and other neurological diseases.

Design of a New Seismoelectric Logging Instrument.

To increase the accuracy of reservoir evaluation, a new type of seismoelectric logging instrument was designed. The designed tool comprises the invented sonde-structured array complex. The tool includes several modules, including a signal excitation module, data acquisition module, phased array transmitting module, impedance matching module and a main system control circuit, which are interconnected through high-speed tool bus to form a distributed architecture. UC/OS-II was used for the real-time system control. After constructing the experimental measurement system prototype of the seismoelectric logging detector, its performance was verified in the laboratory. The obtained results showed that the consistency between the multi-channel received waveform amplitude and benchmark spectrum was more than 97%. The binary phased linear array transmitting module of the instrument can realize 0° to 20° deflection and directional radiation. In the end, a field test was conducted to verify the tool's performance in downhole conditions. The results of this test proved the effectiveness of the developed seismoelectric logging tool.

The effects of a multi-disciplinary team on sleep quality assessment in mild-to-moderate Alzheimer's disease patients with sleep disorders.

BACKGROUND AND AIMS: Sleep disturbances are a severe problem among patients with Alzheimer's disease (AD). By evaluating sleep quality in mild-to-moderate AD patients, this study aimed to assess the effects of multi-disciplinary team (MDT) in reducing the incidence of adverse reactions of AD patients. The reduction in the incidence of adverse reactions to predict multi-disciplinary team (MDT) treatment effects. METHODS AND RESULTS: This study included 60 mild-to-moderate AD patients with sleep problems when hospitalized in Huzhou Third Municipal Hospital. The patients were randomly distributed into two groups, routine and MDT treatments. The cognitive functions, sleep conditions, and psycho-behavioral symptoms were compared between both the groups. Cognitive function declined significantly between pretherapy and follow-up in the routine treatment group (MMSE: t = -7.961, P < 0.001; MoCA: t = -4.672, P < 0.001). There was a significant decline in drowsiness in the MDT group compared to that in the routine treatment group (χ2 = 4.320, P = 0.038). Sleep quality improved significantly during the follow-up in the MDT treatment group (t = 6.098, P < 0.001). The results of the Hamilton Depression Scale (HAMD) and Hamilton Anxiety Scale (HAMA) among family caregivers (FCGs) demonstrated that MDT treatment could alleviate caregivers' depression (t = -2.867, P = 0.042), and routine treatment can worsen their anxiety (t = 3.258, P = 0.003). CONCLUSION: The MDT treatment method as an effective and meaningful therapy can help mitigate the suffering of patients with AD and FCGs.

Neuroprotective effects of zonisamide on cerebral ischemia injury via inhibition of neuronal apoptosis.

It is known that neuronal apoptosis contributes to pathology of cerebral ischemia injury. Zonisamide (ZNS) has shown anti-apoptosis effects in recent studies. The present study investigated whether the anti-apoptotic effect can account for the neuroprotective action of ZNS on cerebral ischemia. Neuronal cells were maintained under oxygen-glucose deprivation conditions to simulate cerebral ischemia and treated with ZNS simultaneously. The apoptosis of the cells and expression of apoptosis-related proteins were investigated by flow cytometry and western blot analysis, respectively. A cerebral ischemia mouse model was created via middle cerebral artery occlusion, and the mice were treated with ZNS. Neurological deficit scores and infarct volumes of the cerebral ischemia mice were measured. The apoptosis status of the neuronal cells was evaluated by TUNEL staining. In vitro, the ZNS treatment inhibited both the apoptosis of the neuronal cells and apoptosis-related protein expression (caspase-3, caspase-8, and calpain-1) induced by the oxygen-glucose deprivation. The anti-apoptosis effect of ZNS could occur through the blocking of reactive oxygen species. Moreover, ZNS treatment significantly ameliorated neurological deficits and reduced infarct volumes in the cerebral ischemia mice model. In this study, ZNS exerted neuroprotective effects by inhibition of apoptosis in neuronal cells in cerebral ischemia. Therefore, ZNS might be a promising therapy for cerebral ischemia.

Use of the A2DS2 scale to predict morbidity in stroke-associated pneumonia: a systematic review and meta-analysis.

BACKGROUND: This review aims to evaluate the performance and clinical applicability of the A2DS2 scale via systematic review and meta-analysis. METHODS: The Medline, Embase, Cochrane Library, CBM, CNKI, and Wanfang databases were searched. The risk of bias was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2). Funnel plots and Egger's test were used to evaluate publication bias. The bivariate random-effect model was used for calculating the sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, and area under the curve (AUC). A Fagan nomogram was applied to evaluate the clinical applicability of the A2DS2 scale. RESULTS: A total of 29 full-text articles met the inclusion criteria, including 19,056 patients. Bivariate mixed-effects regression models yielded a mean sensitivity of 0.78 (95 % CI: 0.73-0.83), a specificity of 0.79 (95 % CI: 0.73-0.84), a positive likelihood ratio of 3.7 (95 % CI: 2.9-4.6), and a negative likelihood ratio of 0.27 (95 % CI: 0.23-0.33). The area under the receiver operating characteristic curve was 0.85 (95 % CI: 0.82-0.88). If given a pre-test probability of 50 %, the Fagan nomogram showed that when A2DS2 was positive, the post-test probability improved to 79 %. In contrast, when A2DS2 was negative, it decreased to 22 %. The results of the subgroup analysis showed no effect on the diagnostic accuracy of the A2DS2 scale in predicting stroke-associated pneumonia, except for the optimal cut-off value. CONCLUSIONS: The A2DS2 scale demonstrates high clinical applicability and could be a valid scale for the early prediction of stroke-associated pneumonia in stroke patients.

Association of air pollution and greenness with carotid plaque: A prospective cohort study in China.

Previous studies indicated that exposure to air pollution was associated with the progress of atherosclerosis, but evidence is very limited in China and even in the world. This study aims to assess the associations of long-term exposures to air pollution and greenness with the occurrence of carotid plaque. Participants of this cohort study were urban residents and office workers who visited Hebei General Hospital for routine physical examination annually from September 2016 through to December 2018. Eligible participants were people diagnosed the absence of carotid plaque clinically at their first hospital visit and were followed up at their second or third hospital visit. Exposure to particulate matter with aerodynamic diameter less than 2.5 µm (PM2.5), nitrogen dioxide (NO2) and ozone (O3) were estimated using an inverse distance weighted (IDW) method. The level of greenness was assessed using the Normalized Difference Vegetation Index (NDVI) and Enhanced Vegetation Index (EVI). The associations were evaluated using Cox proportional hazards regression models. Among 4,137 participants, 575 showed the occurrence of carotid plaque during the follow-up period. After controlling for potential confounders, the hazard ratios (HRs) and 95% confidence intervals (95%CIs) of carotid plaque associated with per interquartile range (IQR) increase in PM2.5, NO2, and O3 were 1.78 (1.55, 2.03), 1.32 (1.14, 1.53) and 1.99 (1.71, 2.31), respectively. Increased EVI and NDVI were significantly associated with lower risk of carotid plaque [HR (and 95%CI): 0.84 (0.77, 0.93) and 0.87 (0.80, 0.94)]. PM2.5 significantly mediated 80.47% or 93.00% of the estimated association between EVI or NDVI and carotid plaque. In light of the significant associations between air pollution, greenness and carotid plaque in this study, continued efforts are needed to curb air pollution and plan more green space considering their effects on vascular disease.

The association between ambient air pollution and blood lipids: A longitudinal study in Shijiazhuang, China.

BACKGROUND: Few studies have explored the associations between ambient air pollution and blood lipid levels. This study aimed to fill this knowledge gap based on a routine health examination cohort in Shijiazhuang, China. METHODS: We included 7063 participants who took the routine health examination for 2-3 times at Hebei General Hospital from January 2016 to December 2018. Individual serum levels of cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured. Their three-month average exposure to air pollution prior to the routine health examinations was estimated using inverse distance weighted method. We used linear mixed-effects regression models to examine the associations between air pollution and levels of blood lipids while controlling for age, gender, body mass index (BMI), smoking, alcohol drinking, temperature, humidity, with a random effect for each individual. RESULTS: Particles with diameters ≤2.5 µm and ≤10 µm (PM2.5 and PM10), nitrogen dioxide (NO2), sulfur dioxide (SO2) and ozone (O3) were all positively associated with TC, TG, and LDL-C and negatively associated with HDL-C, in single pollutant models. Each 10 µg/m3 increment of 3-month average PM2.5 was associated with 0.65% [95% confidence interval (CI): 0.03%-1.28%], 0.56% (95%CI: 0.33%-0.79%) and 0.63% (95%CI: 0.35%-0.91%) increment in TG, TC, and LDL-C, and 0.91% (95%CI: 0.68%-1.13%) decrease in HDL-C. In two-pollutant models, the effects of gaseous pollutants on blood lipids were weakened, while those of PMs were strengthened. Stronger associations were presented in the elderly (≥60 years) and overweight/obese (BMI ≥ 24) participants. CONCLUSIONS: Ambient air pollution had significantly adverse effects on blood lipid levels, especially in overweight/obese and elderly individuals. CAPSULE: Significant associations between increased air pollution and worse blood lipid levels were found, especially in overweight/obese and elderly individuals.

Dl-3-n-butylphthalide promotes neurite outgrowth of primary cortical neurons by Sonic Hedgehog signaling via upregulating Gap43.

Neurite outgrowth is the basis for wiring during the development of the nervous system. Dl-3-n-butylphthalide (NBP) has been recognized as a promising treatment to improve behavioral, neurological and cognitive outcomes in ischemic stroke. However, little is known about the effect and mechanism of NBP on the neurite outgrowth. In this study, we used different methods to investigate the potential effects of NBP on the neurite extension and plasticity of immature and mature primary cortical neurons and explored the underlying mechanisms. Our results demonstrated that in immature and mature cortical neurons, NBP promoted the neurite length and intersections, increased neuritic arborization, elevated numbers of neurite branch and terminal points and improved neurite complexity and plasticity of neuronal development processes. Besides, our data revealed that NBP promoted neurite extension and branching partly by activating Shh signaling pathway via increasing Gap43 expression both in immature and mature primary cortical neurons. The present study provided new insights into the contribution of NBP in neuronal plasticity and unveiled a novel pathway to induce Gap43 expression in primary cortical neurons.

Neuroprotective effects of zonisamide on cerebral ischemia injury via inhibition of neuronal apoptosis

It is known that neuronal apoptosis contributes to pathology of cerebral ischemia injury. Zonisamide (ZNS) has shown anti-apoptosis effects in recent studies. The present study investigated whether the anti-apoptotic effect can account for the neuroprotective action of ZNS on cerebral ischemia. Neuronal cells were maintained under oxygen-glucose deprivation conditions to simulate cerebral ischemia and treated with ZNS simultaneously. The apoptosis of the cells and expression of apoptosis-related proteins were investigated by flow cytometry and western blot analysis, respectively. A cerebral ischemia mouse model was created via middle cerebral artery occlusion, and the mice were treated with ZNS. Neurological deficit scores and infarct volumes of the cerebral ischemia mice were measured. The apoptosis status of the neuronal cells was evaluated by TUNEL staining. In vitro, the ZNS treatment inhibited both the apoptosis of the neuronal cells and apoptosis-related protein expression (caspase-3, caspase-8, and calpain-1) induced by the oxygen-glucose deprivation. The anti-apoptosis effect of ZNS could occur through the blocking of reactive oxygen species. Moreover, ZNS treatment significantly ameliorated neurological deficits and reduced infarct volumes in the cerebral ischemia mice model. In this study, ZNS exerted neuroprotective effects by inhibition of apoptosis in neuronal cells in cerebral ischemia. Therefore, ZNS might be a promising therapy for cerebral ischemia.

Circulating miRNA-3552 as a Potential Biomarker for Ischemic Stroke in Rats.

OBJECTIVE: With the growing incidence of ischemic stroke worldwide, there is an urgent need to identify blood biomarkers for ischemic stroke patients. Thus, our aim was to identify potential circulating microRNA (miRNA) as a potential biomarker and to explore its potential mechanism for ischemic stroke in rats. METHODS: The mRNA dataset GSE97537 and miRNA dataset GSE97532 were downloaded from the Gene Expression Omnibus (GEO) GSE97537 including 7 middle cerebral artery occlusion (MCAO) rat brain tissues and 5 sham-operated rat brain tissues GSE97532 including 6 MCAO rat blood samples and 3 sham-operated rat blood samples. Differentially expressed mRNAs and miRNAs with corrected p value ≤ 0.01 and fold change ≥2 or ≤0.05 were identified. To explore potential biological processes and pathways of differentially expressed mRNAs, functional enrichment analysis was performed. The target mRNAs of differentially expressed miRNAs were predicted using DNA Intelligent Analysis (DIANA)-microT tools. The target mRNAs and differentially expressed mRNAs were intersected. RESULTS: 1228 differentially expressed mRNAs in MCAO rat brain tissues were identified. Highly expressed mRNAs were mainly enriched in the inflammatory responses. Nine differentially expressed miRNAs were identified in MCAO rat blood samples. A total of 673 target mRNAs were predicted to significantly bind these differentially expressed miRNAs. Among them, 54 target mRNAs were differentially expressed in MCAO rat blood samples. Enrichment analysis results showed that these 54 target mRNAs were closely related to neurological diseases and immune responses. Among all miRNA-mRNA relationship, miR-3552-CASP3 interaction was identified, indicating that CASP3 might be mediated by miR-3552. Functional enrichment analysis revealed that CASP3 was involved in the apoptosis pathway, indicating that miR-3552 might participate in apoptosis by CASP3. CONCLUSION: Our findings reveal that circulating miR-3552 shows promise as a potential biomarker for ischemic stroke in rats.

Integrated analysis of the proteome and transcriptome in a MCAO mouse model revealed the molecular landscape during stroke progression.

Strokes usually results in long-term disability and death, and they occur worldwide. Recently, increased research on both on the physiopathological mechanisms and the transcriptome during stroke progression, have highlighted the relationship between stroke progression and immunity, with a special focus on inflammation. Here, we applied proteome analysis to a middle carotid artery occlusion (MCAO) mouse model at 0 h, 6 h, 12 h and 24 h, in which proteome profiling was performed with 23 samples, and 41 differentially expressed proteins (DEPs) were identified. Bioinformatics studies on our data revealed the importance of the immune response and particularly identified the inflammatory response, cytokine- cytokine receptor interactions, the innate immune response and reactive oxygen species (ROS) during stroke progression. In addition, we compared our data with multiple gene expression omnibus (GEO) datasets with and without a time series, in which similar pathways were identified, and three proteins, C3, Apoa4 and S100a9, were highlighted as markers or drug targets for stroke; these three proteins were significantly upregulated in the MCAO model, both in our proteomic data and in the GEO database.

Ginsenoside Rb1 Promotes Motor Functional Recovery and Axonal Regeneration in Post-stroke Mice through cAMP/PKA/CREB Signaling Pathway.

The central nervous system (CNS) has a poor self-repairing capability after injury because of the inhibition of axonal regeneration by many myelin-associated inhibitory factors. Therefore, ischemic stroke usually leads to disability. Previous studies reported that Ginsenoside Rb1 (GRb1) plays a role in neuronal protection in acute phase after ischemic stroke, but its efficacy in post-stroke and the underlying mechanism are not clear. Recent evidences demonstrated GRb1 promotes neurotransmitter release through the cAMP-depend protein kinase A (PKA) pathway, which is related to axonal regeneration. The present study aimed to determine whether GRb1 improves long-term motor functional recovery and promotes cortical axon regeneration in post-stroke. Adult male C57BL/6 mice were subjected to distal middle cerebral artery occlusion (dMCAO). GRb1 solution (5 mg/ml) or equal volume of normal saline was injected intraperitoneally for the first time at 24 h after surgery, and then daily injected until day 14. Day 3, 7, 14 and 28 after dMCAO were used as observation time points. Motor functional recovery was assessed with Rota-rod test and grid walking task. The expression of growth-associated protein 43 (GAP43) and biotinylated dextran amine (BDA) was measured to evaluate axonal regeneration. The levels of cyclic AMP (cAMP) and PKA were measured by Elisa, PKAc and phosphorylated cAMP response element protein (pCREB) were determined by western blot. Our results shown that GRb1 treatment improved motor function and increased the expression of GAP43 and BDA in ipsilesional and contralateral cortex. GRb1 significantly elevated cAMP and PKA, increased the protein expression of PKAc and pCREB. However, the effects of GRb1 were eliminated by H89 intervention (a PKA inhibitor). These results suggested that GRb1 improved functional recovery in post-stroke by stimulating axonal regeneration and brain repair. The underlying mechanism might be up-regulating the expression of cAMP/PKA/CREB pathway.

Role of liraglutide in brain repair promotion through Sirt1-mediated mitochondrial improvement in stroke.

Brain repair, especially axonal sprouting, is critical to restore motor function in disabled stroke patients. Liraglutide (LG) is a new kind of long-acting analogue of glucagon-like peptide-1 (GLP-1) and has potential protective effects in stroke. The mitochondria participate in brain repair after cerebral injury. However, the mechanism of the effect of LG on brain repair and its potential influence on mitochondria in stroke remains obscure. Here, in focal cerebral cortical ischemic mice model, LG improved the motor functional recovery and promoted axonal sprouting by restoring the activities of isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, and succinate dehydrogenase. Moreover, LG remarkably increased the cell survival rate and revived the NeuN and GAP-43 levels in cortical neurons under hydrogen peroxide (H2 O2 ) exposure. It was also observed that LG reduced the generation of reactive oxygen species, stabilized the mitochondrial membrane potential, enhanced the levels of adenosine triphosphate, enhanced activities of mitochondrial complex-I, and decreased protein expression levels of fission-1 in H2 O2 -injured cortical neurons. Additionally, LG suppressed the expressions of sirtuin 1 (Sirt1) in cortical neurons exposed to H2 O2 . Furthermore, knockdown of Sirt1 by short interfering RNA facilitated the LG-mediated mitochondrial protection in cortical neurons under H2 O2 . Collectively, this data from the present study illustrated that LG exerted a promoting influence on brain repair, after cerebral ischemic injury, through Sirt1-mediated mitochondrial improvement.

Tert-butylhydroquinone enhanced angiogenesis and astrocyte activation by activating nuclear factor-E2-related factor 2/heme oxygenase-1 after focal cerebral ischemia in mice.

Angiogenesis after cerebral ischemia plays a pivotal role in neurological recovery and represents a therapeutic target. The angiogenic effect of nuclear factor E2-related factor 2 (Nrf2) was identified in recent years. However, the effects of tert-butylhydroquinone, an Nrf2 inducer, on angiogenesis and astrocyte activation after stroke remain unclear. In this study, we investigated whether tert-butylhydroquinone enhanced angiogenesis and astrocyte activation through Nrf2 pathway. Wild-type (Nrf2+/+) and Nrf2-deficient (Nrf2-/-) mice were subjected to permanent distal middle cerebral artery occlusion (dMCAO). We established 6 experimental groups (sham Nrf2+/+ group, vehicle Nrf2+/+ group, tBHQ Nrf2+/+ group; sham Nrf2-/- group, vehicle Nrf2-/- group, and tBHQ Nrf2-/- group). The infarct volume, neurological function, microvessel density (MVD), astrocytic endfeet covered ratio and the expression of Nrf2, HO-1 and VEGF in the ischemic brain were measured at different time points. Compared with that observed in the vehicle Nrf2+/+ group, tBHQ significantly reduced the infarct volume, enhanced post-stroke angiogenesis and astrocytic endfeet covered ratio in the peri-infarct area. The Nrf2/HO-1/VEGF pathway was activated by tBHQ in the angiogenesis process. However, in Nrf2-/- mice, Nrf2 deficiency blocked the effects of tBHQ on angiogenesis process and neurological recovery as well as abolished the mediation of proangiogenic factors. These results suggested that tBHQ enhanced angiogenesis and astrocyte activation through activating Nrf2 pathway after cerebral ischemia.

Rosuvastatin improves neurite extension in cortical neurons through the Notch 1/BDNF pathway.

Objectives: Neurite outgrowth of neurons is essential for forming functional neural circuits. It is believed that neuronal neurite outgrowth is an important mechanism of brain plasticity. Rosuvastatin (RSV) is a relatively new statin and may have neuroprotective properties. However, whether RSV exerts an effect on neurite extension and its potential mechanism in cortical neurons remains poorly documented. Methods: Immunofluorescence method was used to examine the effect of RSV on neurite outgrowth in primary cortical neuron by measuring neurite length and confirmed the promotion effect. Then, the potential mechanisms involving the Notch1 pathway were investigated. Effects of RSV on the expression of Notch 1 and Hes1were determined using qRT-PCR. In addition, brain-derived neurotrophic factor (BDNF) expression was also assessed using qRT-PCR, and ELISA. Results: RSV promoted neurite outgrowth of cortical neurons, and this effect could be partially prevented by the Notch 1 pathway inhibitor, DAPT. Subsequently, we found that Jagged 1 and Notch 1 were colocalized. In addition, we observed that the levels of both Notch 1 and Hes 1 in cortical neurons were increased after RSV, but sharply decreased after DAPT treatment. Moreover, RSV increased brain-derived neurotrophic factor (BDNF) levels in cortical neurons, but in the culture medium, and the effect could be partially suppressed by DAPT treatment. Discussion: These findings indicate that RSV mediates neurite outgrowth in primary cortical neurons. The RSV-induced neuritogenic effect is mediated at least partly via the Notch1/BDNF pathway.