不同质子泵抑制剂对冠脉支架植入术后氯吡格雷联合阿司匹林抗血小板作用的研究

目的:通过比较奥美拉唑和泮托拉唑对冠状动脉支架术(PCI)后患者血小板功能指标和主要不良心血管事件与出血并发症发生情况,探讨不同质子泵抑制剂对PCI后氯吡格雷联合阿司匹林抗血小板作用的影响。方法:60例实施PCI后常规联合抗血小板治疗(氯吡格雷75mg/d+阿司匹林100mg/d)患者随机分为奥美拉唑组(40mg/d,20例),泮托拉唑组(40mg/d,20例)和对照组(20例),连续用药30d。分别在服药前1d及服药15d,30d用血栓弹力图检测ADP途径诱导的血小板抑制率值和比浊法检测ADP途径诱导的血小板最大聚集率(MPAR)。并观察30d各组主要不良心血管事件和出血并发症的发生情况。结果:①奥美拉唑组和泮托拉唑组与对照组相比,服药前1d及服药15d,30d用血栓弹力图检测的血小板抑制率和比浊法检测的血小板最大聚集率(MPAR)均无明显变化;奥美拉唑与泮托拉唑组间比较,差异也无统计学意义。服药15d,30d与服药前1d相比,每组血小板抑制率明显升高,血小板最大聚集率明显下降,差异有统计学意义(P0.05);但15d和30d相比较,差异无统计学意义。②三组比较心血管事件发生率相近,差异无统计学意义(P0.05);奥美拉唑组和泮托拉唑组比较,心血管事件发生率也无统计学差异(P0.05)。③与对照组比较,奥美拉唑组和泮托拉唑组胃肠道出血发生率均明显减少,有统计学意义(P0.05),但两服药组间比较,出血发生率无明显区别,差异无统计学意义(P0.05)。结论:氯吡格雷联合阿司匹林具有增强血小板抑制,降低血小板凝聚的作用,而不同机制质子泵抑制剂奥美拉唑与泮托拉唑对PCI术后氯吡格雷联合阿司匹林抗血小板治疗患者的血小板功能无明显影响,不降低对心血管事件的预防效果,同时明显降低患者胃肠出血事件的发生率。     

莫诺苷对大鼠局灶性脑缺血再灌注损伤神经功能的影响

目的观察中药山茱萸的有效成分莫诺苷对大鼠局灶性脑缺血再灌注后神经功能的影响。方法Wistar雄性大鼠随机分为假手术组、模型组、莫诺苷小剂量组（30 mg/kg）、莫诺苷中剂量组（90 mg/kg）、莫诺苷大剂量组（270 mg/kg）、维生素E（VE）（35 mg/kg）,采用线栓法制作大鼠局灶性脑缺血再灌注模型,缺血30 min后再灌注3 d,应用Zea Longa法、爬网格、平行木、吊绳、Ludmila Belayev 12分评分法,观察莫诺苷对神经功能缺损的改善作用。结果与模型组比较,莫诺苷给药组（小、中、大剂量）Zea Longa法评分均差异极显著（P〈0.01）;与模型组比较,莫诺苷给药组（小、中、大剂量）吊绳法评分均差异极显著（P〈0.01）;Ludmila Belayev 12分评分法评分与模型组比较,莫诺苷大剂量组差异极显著（P〈0.001）,中剂量组差异极显著（P〈0.01）。结论莫诺苷对局灶性脑缺血再灌注模型大鼠有改善行为学评分的作用。     

纯氧或空气环境复苏对新生大鼠缺氧性大脑皮质神经元超微结构的影响

目的应用缺氧动物模型比较纯氧环境(pure oxygen environment,POE,100%氧)与空气环境(roomair environment,RAE,21%氧)复苏对新生大鼠缺氧性大脑皮质神经元超微结构的影响。方法7日龄20只SD(Sprague Dawley)乳鼠建立缺氧2.5 h模型后,分为纯氧环境组(POE)和空气环境组(RAE),每组再根据复氧后时间点分为2个亚组,即24 h组和72 h组,每亚组5只。按时间点取各组乳鼠大脑半球右侧额顶皮质行电镜样品制备,透射电镜观察。结果复氧各组均可见神经元、神经毡和细胞间隙水肿。RAE 24 h组神经元核膜结构不清,细胞器减少,线粒体肿胀、空泡化、嵴断裂;粗面内质网扩张,常呈空泡状,核糖体减少;高尔基复合体囊泡扩张;溶酶体较多。RAE 72 h组细胞器改变同前,但类似凋亡的细胞核较多,坏死细胞亦较其他组多见。POE 24 h组病变较RAE 24 h组轻。POE 72 h组细胞内线粒体及粗面内质网较丰富,病变亦比RAE 72 h组轻。结论POE复苏较RAE复苏可更能缓解缺氧致神经元超微结构的损伤、减少细胞凋亡及减轻脑水肿。提示,纯氧环境对缺氧复苏后大脑皮质神经元有一定的保护作用,并表明本动物模型适合缺氧新生大鼠大脑皮质神经元研究。     

Dental pulp stem cell‐derived exosomes alleviate cerebral ischaemia‐reperfusion injury through suppressing inflammatory response

ObjectivesThe study aimed to determine whether dental pulp stem cell‐derived exosomes (DPSC‐Exos) exert protective effects against cerebral ischaemia‐reperfusion (I/R) injury and explore its underlying mechanism.Materials and MethodsExosomes were isolated from the culture medium of human DPSC. Adult male C57BL/6 mice were subjected to 2 hours transient middle cerebral artery occlusion (tMCAO) injury followed by 2 hours reperfusion, after which singular injection of DPSC‐Exos via tail vein was administrated. Brain oedema, cerebral infarction and neurological impairment were measured on day 7 after exosomes injection. Then, oxygen‐glucose deprivation–reperfusion (OGD/R) induced BV2 cells were studied to analyse the therapeutic effects of DPSC‐Exos on I/R injury in vitro. Protein levels of TLR4, MyD88, NF‐κB p65, HMGB1, IL‐6, IL‐1β and TNF‐α were determined by western blot or enzyme‐linked immunosorbent assay. The cytoplasmic translocation of HMGB1 was detected by immunofluorescence staining.ResultsDPSC‐Exos alleviated brain oedema, cerebral infarction and neurological impairment in I/R mice. DPSC‐Exos inhibited the I/R‐mediated expression of TLR4, MyD88 and NF‐κB significantly. DPSC‐Exos also reduced the protein expression of IL‐6, IL‐1β and TNF‐α compared with those of the control both in vitro and in vivo. Meanwhile, DPSC‐Exos markedly decreased the HMGB1 cytoplasmic translocation induced by I/R damage.ConclusionsDPSC‐Exos can ameliorate I/R‐induced cerebral injury in mice. Its anti‐inflammatory mechanism might be related with the inhibition of the HMGB1/TLR4/MyD88/NF‐κB pathway.     

大鼠脑缺血再灌注后Cathepsin D和caspase-9表达的动态变化

目的:探讨大鼠缺血再灌注后溶酶体组织蛋白酶D(Cathepsin D CD)、caspase-9在不同时段蛋白质及mRNA表达变化。方法:将60只S D大鼠随机分为三组:正常组(10只),假手术组(10只),脑缺血再灌注组(40只),线栓法制备大脑中动脉梗死模型(MCAO),免疫组化及RT-PCR法分别检测Cathepsin D、caspase-9的蛋白和mRNA表达。结果:与正常组和假手术组比较,模型组大鼠脑缺血再灌注损伤后6h Cathepsin D的蛋白和mRNA表达明显增强(P<0.05),24h达高峰,48h仍保存高水平。caspase-9蛋白和mRNA6h开始明显升高,12h达高峰,此后缓慢下降,但48h组仍显著高于对照组(P<0.05),结论:Cathepsin D、caspase-9在大鼠脑缺血再灌注后表达增强,溶酶体可能参与了脑缺血再灌注损伤后神经细胞凋亡的过程。     

A derivative of the CRMP2 binding compound lanthionine ketimine provides neuroprotection in a mouse model of cerebral ischemia

Lanthionines are novel neurotrophic and neuroprotective small molecules that show promise for the treatment of neurodegenerative diseases. In particular, a recently developed, cell permeable lanthionine derivative known as LKE (lanthionine ketimine 5-ethyl ester) promotes neurite growth at low nanomolar concentrations. LKE also has neuroprotective, anti-apoptotic, and anti-inflammatory properties. Its therapeutic potential in cerebral ischemia and its mechanisms of neurotrophic action remain to be fully elucidated. Here, we hypothesize that the neuroprotective actions of LKE could result from induction or modulation of CRMP2. We found that treating primary cultured mouse neurons with LKE provided significant protection against t-butyl hydroperoxide-induced neuronal death possibly through CRMP2 upregulation. Similarly, in vivo studies showed that LKE pre and/or post-treatment protects mice against permanent distal middle cerebral artery occlusion (p-MCAO) as evidenced by lower stroke lesions and improved functional outcomes in terms of rotarod, grip strength and neurologic deficit scores in treated groups. Protein expression levels of CRMP2 were higher in brain cortices of LKE pretreated mice, suggesting that LKE’s neuroprotective activity may be CRMP2 dependent. Lower activity of cleaved PARP-1 and higher activity of SIRT-1 was also observed in LKE treated group suggesting its anti-apoptotic properties. Our results suggest that LKE has potential as a therapeutic intervention in cerebral ischemia and that part of its protective mechanism may be attributed to CRMP2 mediated action and PARP-1/SIRT-1 modulation.     

Disruption of NMDAR–CRMP-2 signaling protects against focal cerebral ischemic damage in the rat middle cerebral artery occlusion model

Collapsin response mediator protein 2 (CRMP-2), traditionally viewed as an axon/dendrite specification and axonal growth protein, has emerged as nidus in regulation of both pre- and post-synaptic Ca²⁺ channels. Building on our discovery of the interaction and regulation of Ca²⁺ channels by CRMP-2, we recently identified a short sequence in CRMP-2 which, when appended to the transduction domain of HIV TAT protein, suppressed acute, inflammatory and neuropathic pain in vivo by functionally uncoupling CRMP-2 from the Ca²⁺ channel. Remarkably, we also found that this region attenuated Ca²⁺ influx via N-methylD-Aspartate receptors (NMDARs) and reduced neuronal death in a moderate controlled cortical impact model of traumatic brain injury (TBI). Here, we sought to extend these findings by examining additional neuroprotective effects of this peptide (TAT-CBD3) and exploring the biochemical mechanisms by which TAT-CBD3 targets NMDARs. We observed that an intraperitoneal injection of TAT-CBD3 peptide significantly reduced infarct volume in an animal model of focal cerebral ischemia. Neuroprotection was observed when TAT-CBD3 peptide was given either prior to or after occlusion but just prior to reperfusion. Surprisingly, a direct biochemical complex was not resolvable between the NMDAR subunit NR2B and CRMP-2. Intracellular application of TAT-CBD3 failed to inhibit NMDAR current. NR2B interactions with the post synaptic density protein 95 (PSD-95) remained intact and were not disrupted by TAT-CBD3. Peptide tiling of intracellular regions of NR2B revealed two 15-mer sequences, in the carboxyl-terminus of NR2B, that may confer binding between NR2B and CRMP-2 which supports CRMP-2''s role in excitotoxicity and neuroprotection.     

Functional outcome of pannexin-deficient mice after cerebral ischemia

Pannexin (Px, Panx) channels have been implicated in several physiological and pathological processes. We recently studied the potential contribution of pannexins in ischemic brain damage using Px1^-/- Px2^-/- mice and provided evidence that (1) the release of IL-1β and hemichannel function in astrocytes are, in contrast to published data, not affected by the absence of Px1 and Px2, (2) channel function in neurons lacking Px1 and Px2 is impaired and (3) Px1^-/- Px2^-/- mice had a better functional outcome and smaller infarcts than wild-type mice when subjected to ischemic stroke. Here, we further investigate the neurological outcome of wild-type and pannexin double-knockout mice 48 h after permanent occlusion of the distal middle cerebral artery (MCAO). Pannexin double-knockout mice (Px1^-/- Px2^-/-) were less impaired in parameters such as exploration, anxiety, sensorimotor function and behavioral symmetry.     

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 ¹. Because of the ease of genetic manipulation in mice, these models have also been developed in this species ^2-3.Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume ⁴. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.     

显微镜下经外侧裂清除高血压壳核血肿的手术体会

目的:探讨显微镜下经外侧裂-岛叶入路治疗30例高血压壳核出血的手术体会。方法:选择高血压壳核出血患者30例,采取经翼点-外侧裂-岛叶入路显微手术治疗高血压壳核出血的方法,观察记录上述患者手术一般资料及手技体会、手术治疗临床效果及术后第6个月患者的恢复情况。结果:上述患者平均输血量93.3±45.6ml,平均手术时间2.5±0.4小时,平均住院时间23.1±3.3天;血肿清除量>90%者18例,80%～70%者10例,<70%者2例;术后第6个月评价上述患者的ADL,Ⅰ级7例(23.3%),Ⅱ级13例(43.3%),Ⅲ级8例(26.7%),Ⅳ级2例(6.7%),Ⅴ级0例(0.0%),临床疗效满意病例20例(66.7%)。结论:经外侧裂显微手术治疗壳核出血,具有手术时间短、患者创伤小、术后恢复快的优点,值得临床推广。