武汉地区大动脉粥样硬化型缺血性脑卒中患者 载脂蛋白E基因多态性分析

目的：分析武汉地区大动脉粥样硬化型（LAA）缺血性脑卒中患者载脂蛋白E（ApoE）基因多态性的分 布。方法：选取武汉大学人民医院神经内科治疗的LAA型缺血性脑卒中患者179例和健康对照者104例为 研究对象。采用聚合酶链反应（PCR）-荧光探针技术检测卒中组和对照组外周血样本,统计并分析基因型和 等位基因频率的差异,分别比较 2 组不同等位基因组的总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白 （HDL）、低密度脂蛋白（LDL）水平。结果：2组均以ε3/ε3基因型频率最多。对照组ε3/ε3基因频率高于卒中 组（P＜0.05）,卒中组ε3/ε4 和ε4/ε4 分布高于对照组（P＜0.05）;卒中组ε4 等位基因频率高于对照组（P＜ 0.05）,ε3 等位基因频率低于对照组（P＜0.05）。性别对于 ApoE 基因型的分布差异无统计学意义（P＞ 0.05）。对照组ε4型TC、TG、LDL水平均高于ε2型和ε3型（P<0.05或P<0.01）;卒中组ε4型TC水平显著高于ε 2型和ε3型,LDL水平显著高于ε2型（均P＜0.01）。结论：武汉地区LAA型缺血性脑卒中患者ApoE的基因 多态性分布与性别无关但影响血脂水平,ε4可能为LAA型缺血性脑卒中的易感因子和危险因素之一。     

Cerebral Amyloid Angiopathy,Subcortical White Matter Disease and Dementia: Literature Review and Study in OPTIMA

Cerebral amyloid angiopathy (CAA) is of increasing clinical and research interest as the ability to detect it and its consequences by neuroimaging in living subjects has advanced. There is also increasing interest in understanding its possible role in the development of intracerebral hemorrhage, Alzheimer's disease (AD) and vascular dementia. In this article, the literature on this subject is reviewed and novel findings relating CAA to subcortical white matter damage in 224 subjects in the Oxford project to Investigate Memory and Ageing (OPTIMA) are reported. The relationship between CAA and subcortical tissue damage in the OPTIMA subjects was found to be critically dependent on ApoE genotype, there being a positive relationship between measures of CAA and subcortical small vessel disease in ApoEε4 carriers and a significant negative relationship in ApoEε2 carriers. These findings draw attention, as have many other studies, to the importance of ApoE genotype as a major risk factor not only for dementia but also for damage to blood vessels in the aging brain.     

Maternal triglyceride levels and newborn weight in pregnant women with normal glucose tolerance.

OBJECTIVE: To determine the predictive value of serum triglyceride levels (TG) for neonatal weight in pregnant women with positive diabetic screening but normal glucose tolerance. RESEARCH DESIGN AND METHODS: We enrolled 180 pregnant Caucasian women with positive diabetic screening. All women underwent a 3-h 100-g oral glucose tolerance test (OGTT) at 27th +/- 4 week of gestation. At the time of OGTT, we measured: fasting plasma glucose, fasting lipids profile and determined ApoE polymorphisms to evaluate the effects on lipid levels. In 83 women with normal glucose tolerance and at term delivery we evaluated the association between maternal serum TG, specific maternal parameters known to affect fetal growth and newborn weight. RESULTS: Based on OGTT, gestational diabetes mellitus (GDM) was diagnosed in 36 women (20%), impaired glucose tolerance (IGT) in 23 (13%), and normal glucose tolerance (NGT) in 121 (67%). Serum TG concentration was significantly higher in women with GDM (2.47 +/- 0.77 mmol/l) as compared with NGT (1.99 +/- 0.64 mmol/l) or IGT (1.98 +/- 0.81 mmol/l) (P < 0.01). ApoE3 allelic frequency was 86%, ApoE2 and ApoE4 were 5 and 9%, respectively. We found no clear-cut association between apoE genotype and serum TG concentration. Macrosomia and LGA newborns were more frequent in IGT than in GDM or NGT (P < 0.01). In the 83 women with positive diabetic screening but normal glucose tolerance who delivered at term, the incidence of LGA infants was significantly higher in those with TG levels higher than the 75th percentile (> 2.30 mmol/l) (21%) than in mothers who had normal TG levels (4.5%) (P < 0.05). Pre-pregnancy BMI (r(2) = 0.067), weight gain during pregnancy (r(2) = 0.062), fasting serum TG (r(2) = 0.09), and 2-h post-OGTT glucose levels (r(2) = 0.044) were all associated with neonatal body weight (all P < 0.05 or less). However, on a multiple regression analysis, only pre-pregnancy BMI (F-test = 7.26, P < 0.01), and fasting serum TG (F-test = 4.07, P < 0.01) were independently associated with birth weight. CONCLUSIONS: Pre-pregnancy BMI and fasting maternal serum TG determined in the last trimester of gestation were independently associated with neonatal birth weight in women with normal glucose tolerance, but positive screening test. TG levels measured in the third trimester of pregnancy are independent of the genetic polymorphism of ApoE.     

外膜炎症诱发载脂蛋白E基因敲除鼠冠状动脉粥样硬化病灶

研究载脂蛋白E基因敲除(载脂蛋白E°)小鼠冠状动脉内粥样硬化病灶的分布、组成与动脉外膜炎症的关系.取载脂蛋白E°小鼠心脏作连续切片,Movat法染色,追踪冠状动脉主干及其心肌内的小分支;寻找病灶,观察病灶内组成,分析其分布规律.复制小鼠股动脉外膜无菌性炎症模型,用免疫组织化学方法检查内膜粘附分子的表达.结果发现,冠状动脉主干内有延伸病灶,在主干以下分支(包括心肌内小分支)内有在原位生成的病灶,在两类病灶相邻的外膜有炎性细胞浸润,外膜炎症面积大于动脉粥样硬化病灶累及的内膜面积,亦发现一些部位血管外有炎性细胞浸润,而尚无病灶形成.原位病灶均发生于心室壁,大的原位病灶多发生在左室壁心肌内、血管分支处和乳头肌附近的冠状动脉分支内.股动脉外膜炎症可诱发内膜表达细胞间粘附分子1和血管细胞粘附分子1,同时伴白细胞的附壁.以上提示血管外膜炎症是小鼠冠状动脉内病灶的一个始动环节.     

脂欣康对ApoE基因敲除小鼠海马形态学及胆碱乙酰转移酶和突触素表达的影响

目的研究脂欣康对ApoE基因敲除(ApoEKO)小鼠海马形态学及脑胆碱乙酰转移酶(ChAT)和突触素(P38)表达的影响。方法以ApoEKO小鼠为研究对象,随机分为ApoEKO组、脂欣康组、联合干预组,分别给予生理盐水、脂欣康、脂欣康+维生素E灌胃,每日一次,连续灌胃60d,以同龄C57BL/6J小鼠作为正常对照组,之后将所有小鼠断头处死,采用HE染色观察各组小鼠海马神经元组织形态学改变,采用免疫组化技术与计算机图像分析技术检测ApoEKO小鼠海马内ChAT和P38的表达。结果与C57BL/6J小鼠相比,ApoEKO组小鼠海马区神经细胞结构破坏,ChAT阳性神经细胞数目与P38阳性染色颗粒明显减少,染色变浅,ChAT与P38平均灰度值明显增高(P<0.05)。干预组的反应结果介于两者之间,联合干预组表达优于脂欣康组。结论脂欣康能够减轻ApoEKO小鼠海马区神经损害,增强ChAT与P38的表达,与维生素E间有协同作用。     

茶多酚对兔颈总动脉血管成形术后再狭窄的影响

为探讨茶多酚对血管成形术后动脉中膜平滑肌增生及胶原增生的影响,以及与组织型纤溶酶原激活物和血管紧张素Ⅱ活性改变的关系,将雄性新西兰白兔30只随机分为对照组、低剂量茶多酚组和高剂量茶多酚组,用球囊导管剥脱右颈总动脉内皮,造成内皮及中膜损伤,分别在术前、术后3、7、11、14、22和28 d采动脉血应用酶联免疫法测血浆组织型纤溶酶原激活物活性及放射免疫法测血管紧张素Ⅱ血清水平,术后28 d处死动物并取右颈总动脉观察动脉中膜平滑肌和胶原增生程度.结果发现,高剂量茶多酚组组织型纤溶酶原激活物血浆活性为0.169±0.067 IU/L,低剂量茶多酚组为0.141±0.043 IU/L,对照组为0.126±0.043 IU/L,高剂量茶多酚组高于对照组和低剂量茶多酚组,差异具有显著性(P<0.05).高剂量茶多酚组血管紧张素Ⅱ血清水平为1 229±283 ng/L,低剂量茶多酚组为1 302±284 ng/L,对照组为1 309±263 ng/L,三组动物术后血管紧张素Ⅱ血清水平比较差异无显著性.高剂量茶多酚组动脉中膜胶原含量为50.1%+5.82%、低剂量茶多酚组为56.7%±2.3%,对照组为62.8%±2.1%,高剂量茶多酚组低于对照组及低剂量茶多酚组,差异具有显著性 (P<0.05);低剂量茶多酚组低于对照组,差异具有显著性(P<0.001).高剂量茶多酚组中膜平滑肌细胞计数为0.022±0.006/μm2,低剂量茶多酚组为0.034±0.008/μm2,对照组为0.033±0.007/μm2,高剂量茶多酚组低于对照组及低剂量茶多酚组,差异具有显著性(P<0.01).结果提示,高剂量茶多酚可提高血管成形术后血浆组织型纤溶酶原激活物活性,对血管紧张素Ⅱ血清水平无显著性影响,可抑制动脉中膜胶原及平滑肌细胞的增生,这可能有助于减轻或预防动脉血管成形术后再狭窄.     

Hyperactive glial cells contribute to axonal pathologies in the spinal cord of Npc1 mutant mice

Niemann‐Pick disease type C1 (NPC1) is a neurodegenerative disease with various progressive pathological features, for example, neuronal loss, dysmyelination, abnormal axon swelling, and gliosis, in the brain. Pathological activation of p38‐mitogen‐activated protein kinase (MAPK) results in hyperphosphorylation of tau protein, which contributes to the development of neurodegenerative diseases. In this study, axonal varicosities or spheroids and presynaptic aggregates in the spinal cord of the Npc1 mutant mice were found from postnatal day (P) 35 onwards, as indicated by the increased hyperphosphorylated neurofilament and synaptophysin immunoreactivity as well as the findings from electron microscopy. However, activities of astrocytes and microglia in the Npc1 mutant spinal cord were progressively increased earlier from P10 onwards, accompanied by increased expression of interleukin‐1β and apolipoprotein E, as well as up‐regulated p38‐MAPK activity and enhanced phosphorylated tau protein, but not cyclin‐dependent kinase 5/p35 complex and glycogen synthase kinase‐3β. Taken together, our data suggest that the axonal pathologies in the Npc1 mutant spinal cord are strongly correlated with the increase of activated glial cells, which produce IL‐1β and ApoE, resulting in the activation of p38‐MAPK signaling pathway and enhanced phosphorylated tau protein. GLIA 2014;62:1024–1040     

Rho-kinase inhibition improves ischemic perfusion deficit in hyperlipidemic mice

Hyperlipidemia is a major cardiovascular risk factor associated with progressive cerebrovascular dysfunction and diminished collateral perfusion in stroke. Rho-associated kinase (ROCK) may be an important mediator of hyperlipidemic vascular dysfunction. We tested the efficacy of acute or chronic ROCK inhibition on the size of dynamic perfusion defect using laser speckle flowmetry in hyperlipidemic apolipoprotein E knockout mice fed on a high-fat diet for 8 weeks. Mice were studied at an age before the development of flow-limiting atherosclerotic stenoses in aorta and major cervical arteries. Focal ischemia was induced by distal middle cerebral artery occlusion (dMCAO) during optical imaging. The ROCK inhibitor fasudil (10 mg/kg) was administered either as a single dose 1 hour before ischemia onset, or daily for 4 weeks. Fasudil decreased both baseline arterial blood pressure and cerebrovascular resistance (CVR) by ∼15%, and significantly improved tissue perfusion during dMCAO. Interestingly, peri-infarct depolarizations were also reduced. Chronic treatment did not further enhance these benefits compared with acute treatment with a single dose. These data show that ROCK inhibition improves CVR and ischemic tissue perfusion in hyperlipidemic mice.     

Hippocampus-specific deficiency in RNA editing of GluA2 in Alzheimer's disease

Adenosine to inosine (A-to-I) RNA editing is a base recoding process within precursor messenger RNA, catalyzed by members of the adenosine deaminase acting on RNA (ADAR) family. A notable example occurs at the Q/R site of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid glutamate receptor subunit GluA2. Abnormally, low editing at this site leads to excessive calcium influx and cell death. We studied hippocampus and caudate samples from Alzheimer's disease (AD) patients and age-matched healthy controls, using direct sequencing and a high accuracy primer-extension technique to assess RNA editing at the Q/R GluA2 site. Both techniques revealed lower, more variable RNA editing in AD, specific to the hippocampus and the GluA2 site. Deficient editing also characterized the hippocampus of apolipoprotein ε4 allele carriers, regardless of clinical diagnosis. In AD, messenger RNA expression of neuronal markers was decreased in the hippocampus, and expression of the Q/R-site editing enzyme ADAR2 was decreased in caudate. These findings provide a link between neurodegenerative processes and deficient RNA editing of the GluA2 Q/R site, and may contribute to both diagnosis and treatment of AD.     

载脂蛋白E基因多态性与芪参益气滴丸治疗气虚血瘀型冠心病的关系

目的探讨载脂蛋白E(ApoE)基因多态性与芪参益气滴丸治疗气虚血瘀型冠心病疗效的关系。方法符合诊断标准的冠心病患者49例,服用芪参益气滴丸1个月。提取白细胞DNA,扩增ApoE基因第4外显子,测序检测多态性位点。结果发现2个有义突变位点,第2888位突变与气短、神疲乏力的治疗效果有关;第2903位突变对胸痛、自汗的治疗效果有显著影响。结论ApoE基因多态性与芪参益气滴丸治疗气虚血瘀型冠心病证候的疗效有相关性。