1、内源性调节性T细胞在SAH后脑血管痉挛及脑损伤中的作用,汇报人：张金卉导师：孙保亮,2018/9/16,研究的目的及意义,SAH后脑血管痉挛、炎症反应及神经功能预后的改善是 一个非常复杂的病理 、生理、临床过程。虽然进行了大量研究， 为止其 发生机制仍未完全明了。研究小鼠体内内源性调节性T细 对SAH后脑血管痉挛、炎症反应及神经功能预后的改善仍 将是今后一个时期的热点。随着研究的逐渐深入，将 会对内源性调节性T细胞移植SAH的临床治疗及神经功能预后带来 深远的影响。,2018/9/16,国内外相关文献,The Kinetics of Lymphocyte Subsets and Macroph
2、ages in Subarachnoid Space After Subarachnoid Hemorrhage in Rats,it has been suggested that humoral immunity plays a role in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage, there has been no quantitative assay for cellular immunity. We studied the kinetics of immune cells in th
3、e subarachnoid space after subarachnoid hemorrhage in the rat.,2018/9/16,A serial response of immunoreactive cells, which resembles that of the chronic allergic reaction observed in autoimmune diseases or delayed-type hypersensitivity, exists in the subarachnoid space after subarachnoid hemorrhage.
4、The present results suggest that the initial response in cellular immunity, which is followed by humoral immunity and eicosanoid reactions, plays a role in eliciting the development of cerebral vasospasm.,2018/9/16,蛛网膜下腔出血后脑血管痉挛 分子机制的研究进展,2018/9/16,与NO代谢无关的CVS,2018/9/16,Statin-Induced T-Lymphocyte M
5、odulation and Neuroprotection Following Experimental Subarachnoid Hemorrhage,Statins in fl uence immune system activities through echanisms independent of their lipidlowering properties. T cells can be subdivided based on cytokine secretion patterns into two subsets: T-helper cells type 1 (Th1) and
6、type 2 (Th2). Independent laboratory studies have shown statins to be potent inducers of a Th2 switch in immune cell response and be neuroprotective in severalmodels of central nervous system (CNS) disease. This study was the fi rst to evaluate the immune modulating effects of statins in subarachnoi
7、d hemorrhage (SAH).,2018/9/16,The present study elucidated the potential role of a Th2 immune switch in statin provided neuroprotection following SAH.,2018/9/16,Matrix metalloproteinase-9 concentration in the cerebral extracellular fluid of patients during the acute phase of aneurysmal subarachnoid
8、hemorrhage,Elevated cerebral interstitial pro-MMP-9 relates to early brain injury in aSAH patients. A larger prospective study should be performed to confirm whether patients with prolonged elevation or a second peak of cerebral pro-MMP-9 would be more likely to develop DCI and to confirm whether MM
9、P-9, a mediator of neurovascular injury, would be worth to consider as a predictor of vasospasm.,2018/9/16,Trehalose treatment suppresses inflammation,oxidative stress, and vasospasm induced by experimental subarachnoid hemorrhage,Subarachnoid hemorrhage (SAH) frequently results in several complicat
10、ions, including cerebral vasospasm, associated with high mortality. Although cerebral vasospasm is a major cause of brain damages after SAH, other factors such as inflammatory responses and oxidative stress also contribute to high mortality after SAH.Trehalose is a non-reducing disaccharide in which
11、 two glucose units are linked by ,-1,1-glycosidic bond, and has been shown to induce tolerance to a variety of stressors in numerous organisms. In the present study, we investigated the effect of trehalose on cerebral vasospasm, inflammatory responses, and oxidative stress induced by blood in vitro
12、and in vivo.,2018/9/16,trehalose has suppressive effects on several pathological vents after SAH,including vasospasm, inflammatory responses, and lipid peroxidation. Trehalose may be a new therapeutic approach for treatment of complications after SAH.,2018/9/16,properties within the cerebrospinal fl
13、uid after subarachnoid hemorrhage in vivo and in vitro,To functionally characterize pro-inflammatory and vasoconstrictive properties of cerebrospinal fluid after aneurysmal subarachnoid hemorrhage (SAH) in vivo and in vitro.,2018/9/16,We functionally characterized inflammatory and vasoactive propert
14、ies of patients CSF after SAH in vivo and in vitro. This pro-inflammatory milieu in the subarachnoid space might play a pivotal role in the pathophysiology of early and delayed brain injury as well as vasospasm development following SAH.,2018/9/16,研究内容,2018/9/16,每项研究内容均分4组1、Tregs删除组2、SAH模型组3、假手术组4、P
15、BS腹腔注射组体重、窝别、喂养方法相同的小鼠随机分组每组1015只小鼠；每只小鼠编号后 按计算机随机表分4组，双盲（研究者、负责资料收集和分析的人员也不了解分组情况） 较好地避免了偏倚。,研究方法 实验动物处理 1、Tregs删除,2018/9/16,2、SAH模型制作,3、假手术：遇到 阻力即停止继续插入，并迅速退出线栓4、PBS腹腔注射：经腹腔注射与0.25mg CD25特异性抗体等体积的PBS。,2018/9/16,一、内源性调节性T细胞对蛛网膜下腔出血（SAH）后脑血管痉挛的检测内容,检测内容,2018/9/16,局部脑血流量（regional cerebral blood flow,
16、 rCBF）检测：颅骨开窗，在立体定向仪控制下，在SAH前及SAH后12小时内（或6小时）用激光多普勒血流计动态测量动物顶叶皮层rCBF。,2018/9/16,基底动脉形态学检测：,2018/9/16,循环内皮细胞检测：于SAH后3天、2周取静脉血标本，用因子相关抗原免疫荧光染色检测循环内皮细胞（circulating endothelial cell, CEC），以反映血管内皮细胞的损伤。,2018/9/16,于SAH后3天、2周，取动物脑底Willis动脉环及其相连的动脉制备匀浆液PCR、Western blot等检测脑动脉匀浆液中一氧化氮合酶（nitric oxide synthase,
17、 NOS）mRNA和蛋白含量硝酸酶还原法测定NO含量放射免疫分析法测定cGMP含量,2018/9/16,二、内源性调节性T细胞移植SAH后神经功能预后的改善作用,2018/9/16,感觉评分,2018/9/16,运动评分,2018/9/16,认知功能障碍评估,数据表示为动物找到沉水的平台所需要的时间（潜伏期）,2018/9/16,三、内源性调节性T细胞移植对SAH后炎症反应的影响,2018/9/16,小结,实施计划,The present study elucidated the potential role of a Th2 immune switch in statin provided
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