MicroRNA-181c provides neuroprotection in an intracerebral hemorrhage model

Apoptosis is an important factor during the early stage of intracerebral hemorrhage.MiR-181 c plays a key regulatory role in apoptosis.However,whether miR-181 c is involved in apoptosis of prophase cells after intracerebral hemorrhage remains unclear.Therefore,in vitro and in vivo experiments were conducted to test this hypothesis.In vivo experiments:collagenase type VII was injected into the basal ganglia of adult Sprague-Dawley rats to establish an intracerebral hemorrhage model.MiR-181 c mimic or inhibitor was injected in situ 4 hours after intracerebral hemorrhage.Neurological functional defects(neurological severity scores)were assessed 1,7,and 14 days after model establishment.Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and western blot assay were conducted 14 days after model establishment.In vitro experiments:PC12 cells were cultured under oxygen-glucose deprivation,and hemins were added to simulate intracerebral hemorrhage in vitro.MiR-181 c mimic or inhibitor was added to regulate miR-181 c expression.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay,luciferase reporter system,and western blot assay were performed.Experimental results revealed differences in miR-181 c expression in brain tissues of both patients and rats with cerebral hemorrhage.In addition,in vitro experiments found that miR-181 c overexpression could upregulate the Bcl-2/Bax ratio to inhibit apoptosis,while inhibition of miR-181 c expression could reduce the Bcl-2/Bax ratio and aggravate apoptosis of cells.Regulation of apoptosis occurred through the phosphoinositide 3 kinase(PI3 K)/Akt pathway by targeting of phosphatase and tensin homolog deleted on chromosome ten(PTEN).Higher miR-181 c overexpression correlated with lower neurological severity scores,indicating better recovery of neurological function.In conclusion,miR-181 c affects the prognosis of intracerebral hemorrhage by regulating apoptosis,and these effects might be directly mediated and regulated by targeting of the PTEN\PI3 K/Akt pathway and Bcl-2/Bax ratio.Furthermore,these results indicated that miR-181 c played a neuroprotective role in intracerebral hemorrhage by regulating apoptosis of nerve cells,thus providing a potential target for the prevention and treatment of intracerebral hemorrhage.Testing of human serum was authorized by the Ethics Committee of China Medical University(No.2012-38-1)on February 20,2012.The protocol was registered with the Chinese Clinical Trial Registry(Registration No.ChiCTR-COC-17013559).The animal study was approved by the Institutional Animal Care and Use Committee of China Medical University(approval No.2017008)on March 8,2017.     

超敏C反应蛋白与自发性脑出血患者血肿量及预后的相关性分析

目的探讨超敏C反应蛋白(hs-CRP)与自发性脑出血患者(SICH)血肿量和预后的相关性。方法选取2016-01-2018-10萍乡市人民医院收治的SICH患者162例。将其分为大量血肿组、小量血肿组,选择同时期江西萍乡市人民医院80例门诊体检者为对照组。检测和比较3组入院时hs-CRP水平,比较预后良好和不良患者的hs-CRP水平,评价血清hs-CRP水平与血肿体积、预后的相关性。结果大量及小量血肿组hs-CRP水平显著高于对照组(P<0.01),且大量血肿组hs-CRP水平显著高于小量血肿组(P<0.05)。预后良好组hs-CRP水平显著低于预后不良组(P<0.05)。SICH患者hs-CRP水平与血肿量大小和GOS评分呈正相关(r=0.452,0.433,P<0.05)。结论SICH患者血清hs-CRP水平显著升高,与血肿量大小和预后明显相关。     

The Potential Role of Neuromodulation in Subarachnoid Hemorrhage

ObjectivesAneurysmal subarachnoid hemorrhage (SAH) continues to be a difficult cerebrovascular disease with limited pharmacologic treatment options. Cerebral vasospasm (CV) and delayed cerebral ischemia (DCI) are leading causes of morbidity and mortality after SAH. Despite the advances in the understanding of its pathophysiology and tremendous efforts to date, nimodipine is currently the sole Food and Drug Administration–approved treatment for patients with SAH, with benefits that are marginal at best. The neuromodulation therapies are promising, especially those that target CV and DCI to improve functional outcomes. The aim of this review is therefore to summarize the available evidence for each type of neuromodulation for CV and DCI, with a special focus on its pathophysiological mechanisms, in addition to their clinical utility and drawbacks, which we hope will lead to future translational therapy options after SAH.Materials and MethodsWe conducted a comprehensive review of preclinical and clinical studies demonstrating the use of neuromodulation for SAH. The literature search was performed using PubMed, Embase, and ClinicalTrials.gov. A total of 21 articles published from 1992 to 2021 and eight clinical trials were chosen.ResultsThe studies reviewed provide a compelling demonstration that neuromodulation is a potentially useful strategy to target multiple mechanisms of DCI and thus to potentially improve functional outcomes from SAH. There are several types of neuromodulation that have been tested to treat CV and DCI, including the trigeminal/vagus/facial nerve stimulation, sphenopalatine ganglion and spinal cord stimulation, transcranial direct electrical stimulation, transcutaneous electrical neurostimulation, and electroacupuncture. Most of them are in the preclinical or early phases of clinical application; however, they show promising results.ConclusionsDCI has a complex pathogenesis, making the unique anatomical distribution and pleiotropic capabilities of various types of neuromodulation a promising field of study. We may be at the cusp of a breakthrough in the use of these techniques for the treatment of this stubbornly difficult disease.     

Dickkopf-1、LINGO-1、caveolin-1在脑出血中的表达水平变化及其与病情严重程度的关系

目的 探讨Dickkopf-1(DKK-1)、LINGO-1、小窝蛋白1(caveolin-1)在脑出血中的表达水平变化及其与病情严重程度的关系。方法 选取2017年5月-2019年5月在本院就诊的脑出血患者70例,少量出血25例,中量出血24例,大量出血21例; 根据美国国立卫生研究院卒中量表(National institutes of health stroke scale,NIHSS)评分评估脑出血患者的病情严重程度其中轻型26例、中型24例、重型20例; 采用多田氏公式计算患者出血量,其中少量25例、中量24例、大量21例; 手术入路通道中临近血肿0.5cm脑组织作为脑出血组,将远隔血肿位置的脑组织作为对照组; 采用免疫组化染色检测相关因子的表达水平。结果 脑出血组DKK-1、LINGO-1、caveolin-1阳性表达率高于对照组(P<0.05); 大量出血患者阳性表达率高于中量和少量出血患者(P<0.05); 中量出血患者DKK-1、LINGO-1、caveolin-1阳性表达率高于少量出血患者(P<0.05)。重型脑出血患者DKK-1、LINGO-1、caveolin-1阳性表达率高于中型和轻型脑出血患者(P<0.05); 中型脑出血患者DKK-1、LINGO-1、caveolin-1阳性表达率高于轻型脑出血患者(P<0.05)。结论 Dickkopf-1、LINGO-1、caveolin-1在脑出血患者脑组织中高表达,并随着患者病情严重程度的加重,Dickkopf-1、LINGO-1、caveolin-1表达水平越高     

血糖变异性与重症脑出血患者神经功能恢复的相关性

目的 探讨血糖变异性对重症急性脑出血患者神经功能恢复的影响及血糖变异性在重症脑出血患者发病时间轴上的表现特点。方法 选取2018年1月1日-2019年7月1日收入河南科技大学第一附属医院重症外科的脑出血患者,根据患者入院30 d后改良Rankin(Modified Rankin scale,mRS)评分将患者分为神经功能恢复良好组(mRS≤2分),和神经功能恢复不良组(mRS>2分)(残疾/死亡),比较2组入院时高血糖、平均血糖、血糖标准差、入院24 h内血糖变异性(CV1)、入院1～3 d血糖变异性(CV2)、入院3～7 d血糖变异性(CV3)、入院7 d内血糖平均变异性(CV7)、最低血糖水平以及其他临床资料,应用多变量logistic回归分析确定入院30 d后神经功能恢复的独立预测因素。结果 单因素分析显示年龄、CV1、CV3、CV7、最低血糖水平、血糖标准差是影响重症脑出血患者神经功能恢复的相关因素(P<0.05); 多因素logistic逐步回归模型分析显示血糖标准差、CV1、CV3、CV7、最低血糖水平能独立预测重症脑出血患者神经功能的恢复情况; 神经功能恢复良好组和神经功能恢复不良组患者的血糖变异性在入院24 h内、入院1～3 d和入院3～7 d时间轴上的变化特点不同(F=5.000,P=0.029),进一步分析可以看出神经功能恢复不良组的血糖变异性平均幅度较神经功能恢复良好组高,但2组在时间轴上的变化趋势基本相同,均在急性期(入院1～3 d)呈线性上升趋势,之后趋于下降; 组内效应显示患者的血糖变异性在3个时间段上的变化具有显著差异(F=11.663,P<0.001)。结论 血糖标准差、CV1、CV3、CV7、最低血糖水平是影响重症脑出血患者神经功能恢复的独立危险因素; 在重症脑出血患者超急性期、急性期、亚急性期过程中血糖变异性的变化具有显著差异,临床工作中重症脑出血患者应在入院早期密切监测血糖,并积极干预,减小血糖波动范围,以期减少不良预后的发生。     

Suppression of microglial activation and monocyte infiltration ameliorates cerebellar hemorrhage induced-brain injury and ataxia

Ataxia, characterized by uncoordinated movement, is often found in patients with cerebellar hemorrhage (CH), leading to long-term disability without effective management. Microglia are among the first responders to CNS insult. Yet the role and mechanism of microglia in cerebellar injury and ataxia after CH are still unknown. Using Ki20227, an inhibitor for colony-stimulating factor 1 receptor which mediates the signaling responsible for the survival of microglia, we determined the impact of microglial depletion on cerebellar injury and ataxia in a murine model of CH. Microglial depletion reduced cerebellar lesion volume and alleviated gait abnormality, motor incoordination, and locomotor dysfunction after CH. Suppression of CH-initiated microglial activation with minocycline ameliorated cerebellum infiltration of monocytes/macrophages, as well as production of proinflammatory cytokines and chemokine C–C motif ligand-2 (CCL-2) that recruits monocytes/macrophages. Furthermore, both minocycline and bindarit, a CCL-2 inhibitor, prevented apoptosis and electrophysiological dysfunction of Purkinje cells, the principal neurons and sole outputs of the cerebellar cortex, and consequently improved ataxia-like motor abnormalities. Our findings suggest a detrimental role of microglia in neuroinflammation and ataxic motor symptoms after CH, and pave a new path to understand the neuroimmune mechanism underlying CH-induced cerebellar ataxia.     

Carbon monoxide controls microglial erythrophagocytosis by regulating CD36 surface expression to reduce the severity of hemorrhagic injury

Microglial erythrophagocytosis is crucial in injury response to hemorrhagic stroke. We hypothesized that regulation of microglial erythrophagocytosis via HO-1/CO depends on a pathway involving reactive oxygen species (ROS) and CD36 surface-expression. The microglial BV-2 cell line and primary microglia (PMG) were incubated +/−blood and +/−CO-exposure. PMG isolated from tissue-specific HO-1-deficient (LyzM-Cre-Hmox1 ^fl/fl) and CD36 ^−/− mice or siRNA against AMPK (AMP-activated protein kinase) were used to test our hypothesis. In a murine subarachnoid hemorrhage (SAH) model, we compared neuronal injury in wild-type and CD36 ^−/− mice. Readouts included vasospasm, microglia activation, neuronal apoptosis, and spatial memory. We observed increased microglial HO-1-expression after blood-exposure. A burst in ROS-production was seen after CO-exposure, which led to increased amounts of phosphorylated AMPK with subsequently enhanced CD36 surface-expression. Naïve PMG from LyzM-Cre-Hmox1 ^fl/fl mice showed reduced ROS-production and CD36 surface-expression and failed to respond to CO with increased CD36 surface-expression. Lack of HO-1 and CD36 resulted in reduced erythrophagocytosis that could not be rescued with CO. Erythrophagocytosis was enhanced in BV-2 cells in the presence of exogenous CO, which was abolished in cells treated with siRNA to AMPK. CD36 ^−/− mice subjected to SAH showed enhanced neuronal cell death, which resulted in impaired spatial memory function. We demonstrate that microglial phagocytic function partly depends on a pathway involving HO-1 with changes in ROS-production, phosphorylated AMPK, and surface expression of CD36. CD36 was identified as a crucial component in blood clearance after hemorrhage that ultimately determines neuronal outcome. These results demand further investigations studying the potential neuroprotective properties of CO.     

Stellate ganglion ischemia on the prevention of pulmonary vasospasm during bilateral carotid artery ligation: The first experimental study

Fatal pulmonary edema and hemorrhage are significant complications of endovascular treatment in steno-occlusive carotid artery disease; a rational mechanism has not been adequately examined in the literature so far. We investigated if cervical sympathetic ganglia ischemia prevents pulmonary vasospasm on the prognosis of bilateral common carotid artery ligation (BCCAL). Twenty-three adult New Zealand rabbits (4.2 ± 0.3 kg) were randomly divided into three groups: the control group (G1, n = 5), the sham group (G2, n = 6), and the BCCAL group (G3, n = 12). Common carotid arteries were dissected bilaterally in G2/G3, and permanent BCCAL was applied to only in G3. All animals were followed for 3 weeks and decapitated under general anesthesia. Histopathological changes in stellate ganglia and severity of pulmonary vasospasm-related lung edema and hemorrhage were investigated. Results were analyzed by the Kruskal–Wallis test. Two animals of G3 dead within three weeks and the remainder were sacrificed three weeks later. Subpleural petechial foci and an endotracheal bloody fluid collection were grossly observed in the lungs. Histopathologically, pulmonary artery vasospasm, perivascular and subintimal edema, interalveolar hemorrhage, and alveolar wall destructions were observed with less ischemic-degenerated neuron density-determined stellate ganglia animals. Neurodegeneration of stellate ganglia may have a beneficial effect on the prevention of lung injury during steno-occlusive carotid artery disease.     

Identification of intracerebral hemorrhage in the early-phase of MM1+2C-type sporadic Creutzfeldt–Jakob disease: A case report

We report a case of early-phase sporadic Creutzfeldt–Jakob disease (sCJD) complicated by intracerebral hemorrhage (ICH), classified as MM1 + 2C-type based on autopsy. A 61-year-old Japanese man presented to our hospital with speaking difficulties including repeated usage of the same words. He was hospitalized on the seventh day after symptom onset, and diffusion-weighted images on magnetic resonance imaging showed hyperintense regions in the frontal cortex and caudate nucleus. On the 11th day after symptom onset, head computed tomography revealed ICH in the right occipital and parietal lobes. Routine laboratory evaluations and angiography revealed no cause of ICH. Myoclonus of the extremities and drowsiness were observed on the 15th day after symptom onset. He reached the state of akinetic mutism approximately two months after symptom onset. The cerebrospinal fluid test revealed positive real-time quaking-induced conversion and 14-3-3 protein. Electroencephalography revealed periodic sharp wave complexes. A clinical diagnosis of probable Creutzfeldt–Jakob disease was made according to the diagnostic criteria. After a relapse of pneumonia, he passed away on the 103rd day after symptom onset. Postmortem examination revealed ICH in the right posterior cingulate gyrus. No pathological change that might have caused ICH was obtained. Although the effect of sCJD on the onset of ICH is undeniable, the cause of ICH was unknown. Prion protein immunohistochemistry revealed the following results: (1) weak synaptic-type deposits in the tissue rarefacted by ICH; (2) synaptic-type deposits in the cerebral cortex, which showed fine vacuoles; and (3) perivacuolar-type deposits in the inferior temporal gyrus and lingual gyrus, which showed frequent large confluent vacuoles. Although it could be considered MM1-type sCJD clinically, this case was neuropathologically diagnosed as having MM1 + 2C-type sCJD. It was shown that ICH may occur in early-phase sCJD. To improve sCJD prognosis, treatment of complications and careful follow up are important. Furthermore, pathological diagnosis is indispensable for sCJD type diagnosis.