神经干细胞移植治疗脑出血的抗炎作用及其机制研究

发布时间：2018-01-04 02:19

本文关键词：神经干细胞移植治疗脑出血的抗炎作用及其机制研究　出处：《吉林大学》2017年博士论文　论文类型：学位论文

【摘要】：背景与目的:脑出血(Intracerebral hemorahge)是脑卒中的一个亚型,占全部脑卒中的10%-15%,具有很高的发病率、致残率及致死率,对家庭及社会造成极大的影响及沉重的经济负担。脑出血后炎症反应与脑水肿、血脑屏障破坏及病情恶化息息相关。大量动物实验研究表明神经干细胞移植是脑出血的一种较有前景的治疗方法,主要是在抗炎、抗凋亡、促进血管新生等方面发挥作用。然而,其抗炎的具体机制尚不清楚。本研究旨在从体内、体外实验中共同明确神经干细胞治疗脑出血的抗炎作用并在信号传导水平上深入研究其抗炎机制。方法:1、采用胶原酶注射法构建大鼠脑出血模型,造模2小时后通过尾静脉注射植入神经干细胞,观察造模第3天神经干细胞移植对脑组织炎症反应、MAPK信号通路、脑水肿、血脑屏障及行为功能等方面的影响。2、采用溶血产物处理巨噬细胞,构建体外脑出血炎症模型,分别运用三种主要的MAPK通路抑制剂U0126、SP600125、SB203580,探讨这ERK1/2、JNK和p38通路阻断对巨噬细胞炎症反应的影响;通过神经干细胞与巨噬细胞共同培养,在体外探讨神经干细胞在出血性疾病中的抗炎作用及其机制。结果:1、采用立体定位向SD大鼠左侧纹状体内注射0.4U胶原酶构建脑出血模型;脑出血大鼠第3天血肿周围发生了严重的炎症反应,如小胶质细胞/巨噬细胞的激活并伴随有COX-2、IL-1?和TNF-α的表达上调,以及ERK1/2、p38和JNK这三种MAPKs信号通路的激活。神经干细胞移植后能减少血肿周围小胶质细胞/巨噬细胞激活,降低COX-2、IL-1?和TNF-α的表达以及上述三种信号通路的激活,并能减轻脑水肿,保护血脑屏障,促进神经功能恢复。2、溶血产物作用于巨噬细胞后,巨噬细胞胞体增大,细胞表面诸多突起,且炎症介质IL-1?,TNF-α和COX-2的表达均上调且呈动态改变,说明该模型可成功模拟脑出血后炎症反应;溶血产物可迅速激活巨噬细胞内ERK1/2,p38和JNK这三种主要的MAPK信号传导通路,且它们共同参与调控巨噬细胞内炎症反应。神经干细胞与巨噬细胞共培养后能显著抑制溶血产物诱导的巨噬细胞内ERK1/2、p38和JNK信号通路的激活及炎症介质IL-1?,TNF-α和COX-2的表达。结论:本研究证明神经干细胞移植可减轻脑出血后炎症反应,减轻脑水肿及血脑屏障破坏,促进神经功能恢复,并从体内及体外实验同时说明其抗炎作用可能是通过抑制巨噬细胞内ERK1/2、p38和JNK通路激活来实现的。
[Abstract]:Background & objective: intracerebral hemorrhage (ICH) is a subtype of stroke, accounting for 10-15% of all stroke, with a high incidence. The disability rate and fatality rate have a great impact on the family and society and a heavy economic burden. Inflammation and brain edema after cerebral hemorrhage. A large number of animal experiments show that neural stem cell transplantation is a promising treatment for cerebral hemorrhage, mainly in anti-inflammatory, anti-apoptosis. However, the specific mechanism of its anti-inflammatory mechanism is not clear. This study aims at in vivo. In vitro experiments together to clarify the anti-inflammatory effect of neural stem cells in the treatment of intracerebral hemorrhage and to further study its anti-inflammatory mechanism at the level of signal transduction. Methods: 1. The rat model of intracerebral hemorrhage was established by collagenase injection. Neural stem cells were implanted through caudal vein 2 hours after modeling. On the 3rd day, the inflammatory response of brain tissue to MAPK signal pathway and brain edema were observed. Effects of blood-brain barrier and behavioral function. 2. Macrophages were treated with hemolytic products to construct inflammatory model of intracerebral hemorrhage in vitro. U0126, three main inhibitors of MAPK pathway, were used respectively. SP600125 / SB203580 was used to investigate the effects of ERK1 / 2, JNK and p38 pathway blockade on macrophage inflammatory response. The anti-inflammatory effect and mechanism of neural stem cells in hemorrhagic diseases were studied in vitro by co-culture of neural stem cells and macrophages. Results: 1. The model of intracerebral hemorrhage was established by injecting 0.4U collagenase into the left striatum of SD rats by stereotactic localization. Severe inflammatory reactions occurred around hematoma in rats with intracerebral hemorrhage on day 3, such as activation of microglia / macrophages accompanied by COX-2 / IL-1? The expression of TNF- 伪 and TNF- 伪 was up-regulated, as well as ERK1/2. Activation of three MAPKs signaling pathways p38 and JNK. Neural stem cell transplantation can reduce the activation of microglia / macrophage around hematoma and decrease the expression of COX-2 / IL-1. The expression of TNF- 伪 and the activation of the above three signaling pathways can reduce brain edema, protect blood-brain barrier, promote the recovery of nerve function. The hemolytic product acts on macrophages. Macrophage cell body enlarged, cell surface many protrusions, and inflammatory mediator IL-1? The expression of TNF- 伪 and COX-2 were up-regulated and showed dynamic changes, indicating that the model could successfully simulate the inflammatory response after intracerebral hemorrhage. Hemolytic products can rapidly activate three major MAPK signal transduction pathways, ERK1 / 2, p38 and JNK, in macrophages. Neural stem cells co-cultured with macrophages could significantly inhibit ERK1/2 induced by hemolytic products. Activation of p38 and JNK signaling Pathway and Interleukin-1? Conclusion: neural stem cell transplantation can attenuate inflammatory response after cerebral hemorrhage, reduce brain edema and damage of blood-brain barrier, and promote the recovery of nerve function. Both in vivo and in vitro experiments indicated that the anti-inflammatory effect was probably achieved by inhibiting the activation of ERK1 / 2 p38 and JNK pathway in macrophages.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R743.34

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