暨南大学博士学位论文
MALT1蛋白酶活性在脊髓损伤中的免疫调控机制研究
Investigation on immune regulation mechanism of MALT1 paracaspa activity in spinal cord injury
作者姓名：张华
指导教师姓名及学位、职称：高云飞博士教授
学科、专业名称：生物学生物化学与分子生物学
学位类型：学术学位
论文提交日期：
论文答辩日期：2017年5月27日
答辩委员会主席：
论文评阅人：
学位授予单位和日期：
英文爱情故事
中文摘要
外伤性脊髓损伤是一种可以引起持久而严重的感知和运动功能障碍的毁灭性损伤。脊髓损伤后，血-脊髓屏障被破坏则有大量的免疫细胞浸润到损伤部位发挥作用。免疫细胞和炎症因子参与的炎症反应在脊髓损伤后的运动功能修复中至关重要。MALT1（Mucosa-associated lymphoid tissue lymphoma translocation 1）蛋白通过调控免疫细胞的生存和增殖分化在一些神经系统损伤疾病等多种疾病中发挥重要作用，然而MALT1在脊髓损伤中的作用尚未见报道。在本课题中，我们发现MALT1蛋白酶活性缺失或者受到抑制可以促进脊髓损伤后的运动功能的恢复。其中：首先我们在脊髓损伤病人的外周血提取细胞中发现MALT1蛋白酶活性增强；其次，构建MALT1蛋白酶活性缺失小鼠诱导脊髓损伤模型后，发现小鼠运动功能恢复增强，且在损伤部位巨噬细胞的浸润较野生鼠显著减少，除此之外，损伤后，MALT1蛋白酶活性缺失或者受到抑制可以促进损伤部位浸润的巨噬细胞向抑炎巨噬细胞转化，抑制了促炎巨噬细胞方向的转化，从而降低了损伤后24小时内的炎症爆发；我们进行体外实验进一步研究MALT1蛋白酶活性通过调控巨噬细胞来调控脊髓损伤的具体分子机制，发现MALT1蛋白酶活性缺失或者受到抑制（小分子抑制剂MI-2和MLT-827）可以通过抑制NF-κB信号通路的活化来抑制SOCS3的表达进而促进STAT6的活化，从而达到调控巨噬细胞转化的目的；最后，在人的外周血培养的巨噬细胞中应用MALT1蛋白酶活性抑制剂也发现了相同的分子机制以及巨噬细胞相同的转化模式。本研究结果为临床上应用MALT1蛋白酶活性抑制剂来治疗脊髓损伤提供了重要的理论基础。
关键词：MALT1 蛋白酶活性，脊髓损伤，促炎巨噬细胞，抑炎巨噬细胞，NF-κB信号通路，炎症因子
ABSTRACT
Spinal cord injury (SCI) is a devastating traumatic injury that caus persistent, vere motor and nsory dysfunction. After spinal cord injury, the blood-spinal cord barrier is destroyed and a large number of immune cells infiltrate into the injured site, causing an inflammatory respon. Mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1) has been shown to regulate the survival and differentiation of immune cells and plays a critical role in many dias, but its function in lesion recovery after SCI remains unclear. In this paper, deficiency or inhibition of MALT1 paracaspa activity would promote functional recovery after SCI. First, in patients with SCI, PBMC showed incread MALT1 paracaspa activity. Second, we generated KI mice with a point mutation (C472G) in the active center of MALT1 and revealed that the KI mice exhibited improved functional recovery after SCI. Fewer macrophages were recruited to the injury site in KI mice and the macrophages were differentiated into M2 cells. Furthermore, to investigate the specific molecular mechanism by which MALT1 protea activity regulates spinal cord injury by regulating macrophages, we found that macrophages from the KI Mice exhibited reduced phosphorylation of p65, which in turn resulted in decread SOCS3 expression and incread pSTAT3 and pSTAT6 lev
els. Inhibition of MALT1 paracaspa activity with the small molecule inhibitor “MI-2” or the more specific inhibitor “MLT-827” yielded similar results. Last but not least, human macrophages showed a reduced M1 phenotype and a stronger M2 phenotype following the inhibition of MALT1 paracaspa activity. The findings provided important clues suggesting that the inhibition of MALT1 paracaspa activity may improve lesion recovery in subjects with SCI in the clinic.
Keywords: anti-inflammatory macrophage, pro-inflammatory macrophage, spinal cord injury, NF-κB, MALT1 paracaspa activity, inflammatory cytokines
目录
中文摘要............................................................................................................................................ I I 1前言 .. (1)
1.1脊髓损伤 (1)
1.2巨噬细胞 (6)
1.3 MALT1蛋白 (10)
最大的蜗牛
2.实验材料和方法 (18)
2.1 实验小鼠及人体样本 (18)
文襄
2.2 实验材料 (19)
2.3 实验方法 (24)
2.4 数据的统计分析 (33)
在生活方面3.实验结果 (34)
3.1 脊髓损伤病人外周血中MALT1蛋白酶活性发生变化 (34)
3.2 MALT1蛋白酶活性的缺失或受到抑制促进小鼠脊髓损伤后的运动功能恢复 (36)
3.3 骨髓来源细胞的MALT1蛋白酶活性调控脊髓损伤后运动功能的恢复 (46)
3.4 具有MALT1蛋白酶活性的巨噬细胞调控脊髓损伤后的运动功能修复 (49)
3.5 MALT1蛋白酶活性通过巨噬细胞来调控脊髓损伤的运动功能恢复 (52)
长沙大学是几本
3.6 MA TI蛋白酶活性缺失调控体外培养BMDM的极化和炎症因子表达 (57)
3.7 MALT1通过调控NF-κB信号通路和SOCS通路调控巨噬细胞转化 (60)
3.8 脊髓损伤促进病人脑脊液中促炎巨噬细胞的浸润以及促炎因子的表达 (66)
3.9 人的巨噬细胞中NF-κB信号通路的活化同样需要MALT1蛋白酶活性的调控 (69)墨鱼怎么吃
韦鲁斯天赋3.10 MALT1蛋白酶活性可以调控人的巨噬细胞的转化 (71)
4.结论与讨论 (73)
分娩期并发症参考文献 (79)
附录 (89)
在校期间论文发表情况 (96)
致谢 (97)
