小鼠脑缺血再灌注损伤恢复期脂质介导的炎症反应的分子机制研究

DISSECTING LIPID-INDUCED INFLAMMATORY MOLECULAR PATHWAYS DURING BRAIN RECOVERY FOLLOWING MIDDLE CEREBRAL ARTERY OCCLUSION IN MICE

谢承滨

11649083

硕士

生物学

侯圣陶

2018-06-01

2018-07-05

哈尔滨工业大学

深圳

脑卒中已经成为威胁人类生命的最主要疾病之一，正严重威胁着居民的生存、健康和生活质量。而目前对于脑卒中的发病分子机理知之甚少，脑卒中治疗也只能通过通栓药的方式微弱进行。脂质组学作为代谢组学的一个分支，这些年来越来越多地应用在生物医学研究领域，用于相关疾病的生物标志物诊断、揭示疾病的发病分子机制。本论文旨在通过脂质组学技术研究脑卒中诱发的内源性神经脂质变化，找到卒中恢复期关键的脂质分子，探究卒中后神经炎症分子机理，发现保护神经细胞的脂质生物标记物，开发有效治疗脑卒中的新方法。本论文使用脑卒中的疾病动物模型 MCAO 模型，借助高灵敏度的UPLC-MS 分析平台，研究小鼠脑缺血/再灌注损伤恢复期大脑皮层区域不同脂质的变化，建立了大脑脂组代谢图谱，通过建立 PCA 模型和 OPLS-DA 模型对小鼠脑缺血/再灌注 1 天、3 天、7 天、14 天、28 天以及假手术组共 41 个脂质样品里的 578 个脂质分子分析各组间差异脂质，并结合 OPLS-DA 模型中的VIP > 1 及 t 检验的 p < 0.05 共筛选出 84 个主要的差异脂质化合物，通过统计分析 84 个差异脂质分子在恢复期的变化及 MCAO 行为学情况，得到 7 个潜在生物标志物，分别是：PC(20:4/22:6)、PC(16:0/18:2)、PC(16:0/16:0)、LPC(16:0)、PG(18:1/20:4)、PE(16:0p/16:1)和 Cer(d18:2/18:0)。其中 PC(16:0/16:0)和 LPC(16:0)脂质分子在恢复期的变化最显著，且 PC(16:0/16:0)在磷脂酶 A2 的作用下产生LPC(16:0)。通过研究 PC(16:0/16:0)和 LPC(16:0)与小胶质细胞激活的关系，发现PC(16:0/16:0)和 LPC(16:0)均不能使小胶质细胞激活产生炎症因子，通过建立LPS 诱导的小胶质细胞激活分泌 TNF-α、IL-1β 炎症模型，发现 PC(16:0/16:0)能显著降低 LPS 诱导的小胶质细胞分泌的 TNF-α、IL-1β。PC(16:0/16:0)分子作为潜在的神经保护剂对卒中后神经炎症的恢复有重要意义。

Stroke has become one of the most serious diseases that threaten human life and is seriously threatening the survival, health and quality of life of residents. At present, there is little known about the molecular mechanism of stroke and stroke therapy can only be performed in a weak way through the bolt. As a branch of metabonomics, lipidomics has been increasingly used in the field of biomedical research over the past years to diagnose biomarkers of related diseases and reveal the molecular mechanism of the disease. This reasearch aims to explore the changes of endogenous neurolipids induced by stroke using lipidomics technology, find the key lipid molecules in the recovery phase of stroke, explore the molecular mechanism of post-stroke neuroinflammation, and discover the lipid biomarkers that protect nerve cells. To develop new methods for effective treatment of stroke.In this dissertation, mouse MCAO model of cerebral apoplexy disease was used to study the changes of different lipids in the cerebral cortex of mice after cerebral ischemia/reperfusion injury using a high-sensitivity UPLC-MS analysis platformand establish a group of metabolic brain lipid profiles. The establishment of PCA model and OPLS-DA model was used to analyze 578 lipid molecules in 41 lipid samples collected from 1, 3, 7, 14 and 28 days after cerebral ischemia/reperfusion in mice and sham operation group. In combination with the OPS-DA model with VIP > 1 and the t-test, p < 0.05, a total of 84 major lipid compounds were screened out, and 84 changes in lipids during recovery and MCAO behaviors were statistically analyzed. Seven potential biomarkers were obtained: PC(20:4/22:6), PC(16:0/18:2), PC(16:0/16:0), LPC(16:0), PG(18:1/20:4), PE(16:0p/16:1) and Cer(d18:2/18:0). Among them, PC(16:0/16:0) and LPC(16:0) lipid molecules showed the most significant changes during the long time recovery period, and PC(16:0/16:0) produced LPC(16:0) under the action of phospholipase A2.By studying the relationship between PC(16:0/16:0) with LPC(16:0) and microglia activation, we found that neither PC(16:0/16:0) nor LPC(16:0) could make microglia activate and secret inflammatory cytokines. By establishingLPS-induced microglia activation and secretion of TNF-α and IL-1β inflammation models. We found that PC(16:0/16:0) can significantly reduce LPS-induced microglia cells secreted TNF-α, IL-1β. The PC(16:0/16:0) molecule as a potential neuroprotective agent has important implications for the recovery of post-stroke neuroinflammation.

脑卒中 中动脉栓塞模型 脂质组学 液质联用 小胶质细胞 炎症因子

Stroke MCAO model Lipidomics LC-MS Microglia Inflammatory cytokines

中文

联合培养

南方科技大学

谢承滨. 小鼠脑缺血再灌注损伤恢复期脂质介导的炎症反应的分子机制研究[D]. 深圳. 哈尔滨工业大学,2018.