內(nèi)源性調(diào)節(jié)性T細(xì)胞在SAH后腦血管痙攣及腦損傷中的作用ppt課件

1、 內(nèi)源性調(diào)理性?xún)?nèi)源性調(diào)理性T細(xì)胞在細(xì)胞在SAH后腦血后腦血管痙攣及腦損傷中的作用管痙攣及腦損傷中的作用匯報(bào)人：張金卉匯報(bào)人：張金卉 導(dǎo)師：孫保亮導(dǎo)師：孫保亮研討的目的及意義研討的目的及意義SAH后腦血管痙攣、炎癥反響及神經(jīng)功能預(yù)后的改善是一個(gè)非常復(fù)雜的病理 、生理、臨床過(guò)程。雖然進(jìn)展了大量研討，為止其 發(fā)活力制仍未完全明了。研討小鼠體內(nèi)內(nèi)源性調(diào)理性T細(xì)對(duì)SAH后腦血管痙攣、炎癥反響及神經(jīng)功能預(yù)后的改善仍將是今后一個(gè)時(shí)期的熱點(diǎn)。隨著研討的逐漸深化，將會(huì)對(duì)內(nèi)源性調(diào)理性T細(xì)胞移植SAH的臨床治療及神經(jīng)功能預(yù)后帶來(lái)深遠(yuǎn)的影響。國(guó)內(nèi)外相關(guān)文獻(xiàn)國(guó)內(nèi)外相關(guān)文獻(xiàn) The Kinetics of Lymphoc

2、yte Subsets andMacrophages 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

3、 of immune cells in the subarachnoid space after subarachnoid hemorrhage in the rat. 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

4、hemorrhage. 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.蛛網(wǎng)膜下腔出血后腦血管痙攣蛛網(wǎng)膜下腔出血后腦血管痙攣分子機(jī)制的研討進(jìn)展分子機(jī)制的研討進(jìn)展與NO代謝有關(guān)的CVSK 通道活性的改動(dòng)文氧合血紅蛋白的作用本磷酸二酯酶一V的作

5、用本NO和ET一1的平衡失調(diào)與與NO代謝無(wú)關(guān)的代謝無(wú)關(guān)的CVSRas蛋白的作用前列環(huán)素與血栓烷A2失衡自在基與脂質(zhì)過(guò)氧化物增多血管加壓素的作用ca 代謝紊亂Statin-Induced T-Lymphocyte Modulation and Neuroprotection Following Experimental Subarachnoid Hemorrhage Statins in fl uence immune system activities through echanisms independent of their lipidlowering properties. T cell

6、s can be subdivided based on cytokine secretion patterns into two subsets: T-helper cells type 1 (Th1) and 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 (

7、CNS) disease. This study was the fi rst to evaluate the immune modulating effects of statins in subarachnoid hemorrhage (SAH). The present study elucidated the potential role of a Th2 immune switch in statin provided neuroprotection following SAH.Matrix metalloproteinase-9 concentration in the cereb

8、ral extracellular fluid of patients during the acute phase of aneurysmal subarachnoid hemorrhage Elevated cerebral interstitial pro-MMP-9 relates toearly brain injury in aSAH patients. A larger prospectivestudy should be performed to confirm whether patients with prolonged elevation or a second peak

9、 of cerebral pro-MMP-9 would be more likely to develop DCI and to confirm whether MMP-9, a mediator of neurovascular injury, would be worth to consider as a predictor of vasospasm.Trehalose treatment suppresses inflammation,oxidative stress, and vasospasm induced by experimental subarachnoid hemorrh

10、age Subarachnoid hemorrhage (SAH) frequently results in several complications, 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 h

11、igh mortality after SAH.Trehalose is a non-reducing disaccharide in which 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

12、, inflammatory responses, and oxidative stress induced by blood in vitro and in vivo. 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 complicati

13、ons after SAH.properties within the cerebrospinal fluid 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. We functionally charact

14、erized inflammatory and vasoactive properties 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.研討內(nèi)容研討內(nèi)容小鼠體內(nèi)內(nèi)源性調(diào)理性T細(xì)胞

15、對(duì)蛛網(wǎng)膜下腔出血SAH后腦血管痙攣的影響小鼠體內(nèi)內(nèi)源性調(diào)理性T細(xì)胞移植SAH后神經(jīng)功能預(yù)后的改善作用小鼠體內(nèi)內(nèi)源性調(diào)理性T細(xì)胞移植對(duì)SAH后炎癥反響的影響 每項(xiàng)研討內(nèi)容均分4組1、Tregs刪除組2、SAH模型組3、假手術(shù)組4、PBS腹腔注射組體重、窩別、喂養(yǎng)方法一樣的小鼠隨機(jī)分組每組1015只小鼠；每只小鼠編號(hào)后按計(jì)算機(jī)隨機(jī)表分4組，雙盲研討者、擔(dān)任資料搜集和分析的人員也不了解分組情況較好地防止了偏倚。 研討方法研討方法實(shí)驗(yàn)動(dòng)物處置實(shí)驗(yàn)動(dòng)物處置1、Tregs刪除刪除腹腔注射0.25mg CD25特異性抗體小鼠參與肝素抗凝Hanks 液洗滌、ELS裂解注射后第2天取外周血參與標(biāo)志的抗細(xì)胞外表特

16、異性抗原的抗體PBS 洗兩遍4 下避光反響20-30分鐘，洗滌2 次參與標(biāo)志的抗細(xì)胞外表特異性抗原的抗體FACS Calibur 檢測(cè)CD4+CD25+T細(xì)胞300l染色緩沖液重懸細(xì)胞CD4+CD25+T細(xì)胞幾乎減少至0%為調(diào)理性T細(xì)胞刪除勝利Cell quest 搜集細(xì)胞并分析流式結(jié)果2、SAH模型制造模型制造覺(jué)得有阻力時(shí)再繼續(xù)前進(jìn)3mm, 總進(jìn)入長(zhǎng)度約為10mm(已刺破動(dòng)脈壁然后迅速將線栓拔出，動(dòng)脈血灌入，呵斥蛛網(wǎng)膜下腔出血將制備的線栓經(jīng)過(guò)切口經(jīng)頸動(dòng)脈插入頸內(nèi)動(dòng)脈，不斷送至大腦前動(dòng)脈與大腦中動(dòng)脈的分叉處類(lèi)似大腦中動(dòng)脈阻斷的方法首先制造線栓5-0單絲尼龍線，頭端磨成子彈頭狀，總長(zhǎng)度15mm，

17、直徑0.2mm3、假手術(shù)：遇到、假手術(shù)：遇到 阻力即停頓繼續(xù)插入，并迅速退出線栓阻力即停頓繼續(xù)插入，并迅速退出線栓4、PBS腹腔注射：經(jīng)腹腔注射與腹腔注射：經(jīng)腹腔注射與0.25mg CD25特異性抗體特異性抗體等體積的等體積的PBS。 一、內(nèi)源性調(diào)理性一、內(nèi)源性調(diào)理性T細(xì)胞對(duì)蛛網(wǎng)膜下腔出血細(xì)胞對(duì)蛛網(wǎng)膜下腔出血SAH后腦血管痙攣的檢測(cè)內(nèi)容后腦血管痙攣的檢測(cè)內(nèi)容Text基底動(dòng)脈形狀學(xué)檢測(cè)腦動(dòng)脈勻漿液中一氧化氮合酶mRNA和蛋白含量；NO含量；cGMP含量部分腦血流量部分腦血流量循環(huán)內(nèi)皮細(xì)胞檢測(cè)循環(huán)內(nèi)皮細(xì)胞檢測(cè)基底動(dòng)脈形狀學(xué)檢測(cè)腦動(dòng)脈勻漿液中一氧化氮合酶mRNA和蛋白含量；NO含量；cGMP含量部分

18、腦血流量部分腦血流量循環(huán)內(nèi)皮細(xì)胞檢測(cè)循環(huán)內(nèi)皮細(xì)胞檢測(cè)檢測(cè)內(nèi)容部分腦血流量regional cerebral blood flow, rCBF檢測(cè)：顱骨開(kāi)窗，在立體定向儀控制下，在SAH前及SAH后12小時(shí)內(nèi)或6小時(shí)用激光多普勒血流計(jì)動(dòng)態(tài)丈量動(dòng)物頂葉皮層rCBF。基底動(dòng)脈形狀學(xué)檢測(cè)：1、立體顯微鏡下察看基底動(dòng)脈BA、大腦中動(dòng)脈MCA和大腦前動(dòng)脈ACA并記錄它們的直徑2、冰凍切片上顯微鏡下察看基底動(dòng)脈形狀學(xué)變化，用圖像分析儀測(cè)定基底動(dòng)脈內(nèi)徑、外徑與管腔橫截面積改用10明膠和墨水混合液灌注13.5 mlmin)15s于SAH后2天、2周，經(jīng)心臟灌注，10福爾馬林灌注5.5 mlminmin 后循環(huán)內(nèi)

19、皮細(xì)胞檢測(cè)：于SAH后3天、2周取靜脈血標(biāo)本，用因子相關(guān)抗原免疫熒光染色檢測(cè)循環(huán)內(nèi)皮細(xì)胞circulating endothelial cell, CEC，以反映血管內(nèi)皮細(xì)胞的損傷。于SAH后3天、2周，取動(dòng)物腦底Willis動(dòng)脈環(huán)及其相連的動(dòng)脈制備勻漿液 PCR、Western blot等檢測(cè)腦動(dòng)脈勻漿液中一氧化氮合酶nitric oxide synthase, NOSmRNA和蛋白含量 硝酸酶復(fù)原法測(cè)定NO含量 放射免疫分析法測(cè)定cGMP含量二、內(nèi)源性調(diào)理性二、內(nèi)源性調(diào)理性T細(xì)胞移植細(xì)胞移植SAH后神經(jīng)功能后神經(jīng)功能預(yù)后的改善作用預(yù)后的改善作用運(yùn)動(dòng)評(píng)分共運(yùn)動(dòng)評(píng)分共12分分覺(jué)得評(píng)分覺(jué)得評(píng)分515分分認(rèn)知功能妨礙評(píng)價(jià)認(rèn)知功能妨礙評(píng)價(jià)振動(dòng)覺(jué)振動(dòng)覺(jué)13分分觸覺(jué)觸覺(jué)13分分視覺(jué)視覺(jué)13分分 對(duì)刺激的觸激反響對(duì)刺激的觸激反響13分分嗅覺(jué)嗅覺(jué)13分分覺(jué)得評(píng)分運(yùn)動(dòng)評(píng)分 平衡和協(xié)調(diào)才干3分 隨意活 動(dòng)3分 運(yùn)動(dòng)評(píng)分攀爬才干攀爬才干3 3分分 肢體活動(dòng)對(duì)稱(chēng)性3分 認(rèn)知功能妨礙評(píng)價(jià)認(rèn)知功能妨礙評(píng)價(jià)在2次測(cè)試中的后一次測(cè)試中，將平臺(tái)移去，讓動(dòng)物