MMP-9、MMP-2、TIMP-1及TIMP-2在创伤性颅脑损伤患者中的表达及意义

发布时间：2018-04-05 18:21

本文选题：创伤性颅脑损伤　切入点：基质金属蛋白酶　出处：《河北大学》2013年硕士论文

【摘要】：目的：创伤性颅脑损伤（Traumatic brain injury）在西方国家早就引起人们的注意，，它的致死和致残的非常高[1]。改革开放以后这些年，我国的经济的如火箭升天般一样直速上升，汽车的普遍，高速公路的提速等，我国颅脑伤发生率和因颅脑伤致死致残的伤员也逐年增加。因此越来越多的学者开始关注并研究它。创伤性颅脑损伤遗留的偏瘫，智力障碍等重大残疾,给社会和家庭带来沉重负担，脑损伤后的直接结果是形成脑水肿,随着脑水肿的进行性加重,颅内压增高,常导致脑病而死亡,且死亡率非常高，随着相关研究的进一步深入,认识到脑损伤中血脑屏障(BBB)的损害在脑水肿的形成中起十分明显的作用,而血脑屏障损害的实质是一种膜系统受损的结果,对血脑屏障的超微结构研究发现,BBB由三层结构组成:脑毛细血管内皮细胞及其紧密连接而形成的基膜和胶质细胞足突，脑损伤再灌注的损害亦表现为这三种结构的破坏,从而改变其通透性致脑水肿的发生，基质金属蛋白酶(MMPS)是一个比较大的蛋白酶家族,主要分解细胞外基质(ECM),机体同时存在金属蛋白酶的组织抑制物(TIMPS),MMPS与TIMPS之间保持相对的特异性和功能上的动态平衡，在各种病理条件下打破了这种平衡,就会导致过分的细胞外基质分解,产生相应的病理损害。因此通过检测并分析脑外伤后的患者脑脊液及血液中MMP-9、MMP-2、TIMP-1及TIMP-2的表达与正常对照组中它们的表达的差异性，从而来探讨TBI患者的脑脊液及血液中MMP-9、MMP-2、TIMP-1及TIMP-2的表达水平与TBI患者的的严重程度以及临床预后等之间的关系。方法：收集2012年1月～2012年10月期间河北大学附属医院神经外科诊断为ＴＢＩ患者30例,30例患者按（Glasgow≤8分为重度组，8分的为轻度组）分为两组，全部TBI患者于发病24小时内入院,分别留取24h,72h和120h之内脑脊液及静脉血，30例ＴＢＩ患者测定MMP-9、MMP-2、TIMP-1及TIMP-2，对照组于采集时分别留取静脉血及脑脊液各5ml左右,血液标本（脑脊液标本中为血性脑脊液的）静置30分钟使血液凝集，其余标本可直接检验,室温1000Xg离心１０分钟,吸取上清液(血清),立即置于-70℃，冰箱保存待测，ＴＢＩ患者分别分为观察组轻重两组,同时选取10例对照组（标本为检验科赠送,脑脊液标本为排除内科颅内疾病时筛选的正常脑脊液，均经检测为正常，血液标本为健康体检时所采集的标本）,所有采集标本用双抗体夹心ELISA法检测观察因子。结果：本实验检测到MMP-9在TBI患者组脑脊液中24小时内达到高峰，最少可以持续72小时，统计学显示，24小时内与72小时内无显著性差别，但是与120小时内的检测有显著性差别。TIMP-1在120小时内可达高峰，相对第72小时内，第24小时内有显著性差异。MMP-2及TIMP-2在TBI患者的脑脊液中120小时内无明显变化。在血液中各检测因子随时间无明显变化，TBI患者与对照组的MMP-9、MMP-2、TIMP-1及TIMP-2无论CST还是血液中的表达都有显著性差异.重度TBI患者组的MMP-9在脑脊液水平显著高于轻度TBI患者组。然而MMP-2、TIMP-1及TIMP-2在脑脊液中的表达在重度与轻度组之间无显著性差异。在血液标本检测中MMP-9、MMP-2、TIMP-1及TIMP-2的表达在重度与轻度组之间无显著性差异。对所有的TBI患者组受伤后MMP-9（120小时）含量与TIMP-1（120小时）含量进行简单相关性分析，结果显示，两者之间有直线相关关系，呈负相关，相关系数-0.593（P0.01），MMP-2与TIMP-2之间在各检测点保持相关性。结论：本课题在TBI不同分级患者及正常脑组织中检测并分析了MMP-9、MMP-2、TIMP-1及TIMP-2的表达水平。研究发现如下： 1.TBI患者与对照组的MMP-9、MMP-2、TIMP-1及TIMP-2无论脑脊液还是血液中的表达都有显著性差异。MMP-9在TBI患者组脑脊液中24小时内达到高峰，最少可以持续72小时。对TBI患者来说，MMP-9的主要来源可能是神经细胞。 2.MMP-9（120小时）含量与TIMP-1（120小时）在脑脊液中表达的含量进行简单相关性分析，两者变化趋势相反。TIMP-1可能抑制MMP-9。 3.MMP-2与TIMP-2之间在在脑脊液中的表达于各检测点保持负相关性，两者变化趋势相反。TIMP-2可能特异性抑制MMP-2。 4.TIMP-1可能是内生源性保护因子。 5.MMP-9可以作为预测临床预后的指标。
[Abstract]:Objective: traumatic brain injury (Traumatic brain injury) in western countries have long attracted attention after its morbidity and mortality is very high [1]. these years of reform and opening up, China's economy such as the rocket like speed straight rise, the common highway speed, our brain injury the incidence of death and disability due to brain injury and the wounded also increased year by year. Therefore, more and more scholars begin to pay attention to and study it. Traumatic brain injury left hemiplegia, mental retardation and other major disability, bring heavy burden to society and family, a direct result of brain damage is the formation of brain edema, with progressive cerebral edema, increased intracranial pressure, often lead to encephalopathy and death, and the mortality rate is very high, with further research, recognizing the brain damage of blood brain barrier (BBB) damage on edema formation in the very Obviously, the essence of BBB damage is the result of a damaged membrane system, ultrastructural study on blood brain barrier, BBB is composed of three layers: basement membrane and glial cell processes of brain capillary endothelial cells and tight junctions, brain injury and reperfusion injury also show that three kinds of structural damage, so as to change the permeability of brain edema caused by the occurrence of matrix metalloproteinase (MMPS) is a large protein family. The main decomposition of extracellular matrix (ECM), and the body of tissue inhibitor of metalloproteinases (TIMPS), keep the dynamic balance of specificity and relative function between the MMPS and TIMPS broke this balance in a variety of pathological conditions, will lead to excessive extracellular matrix decomposition, produce corresponding pathological damages.
Therefore, through the detection and analysis of blood and cerebrospinal fluid in patients with MMP-9 after traumatic brain injury in MMP-2, the differences of their expression of expression of TIMP-1 and TIMP-2 and the normal control group, so as to explore the MMP-9, blood and cerebrospinal fluid of patients with TBI in MMP-2, the relationship between the severity of the expression level of TIMP-1 and TIMP-2 in patients with TBI. And the clinical prognosis.
Methods: from January 2012 to October 2012 for the diagnosis of the Department of neurosurgery in the Affiliated Hospital of Hebei University during the period of 30 TBI patients, 30 patients (according to Glasgow is less than or equal to 8 divided into severe group, 8 were mild group) were divided into two groups, all TBI patients within 24 hours of admission were collected within 24h, 72h and 120h in cerebrospinal fluid and venous blood were measured in 30 patients with MMP-9, patients with TBI MMP-2, TIMP-1 and TIMP-2 in the control group were collected venous blood and cerebrospinal fluid of all blood samples (about 5ml in cerebrospinal fluid for bloody cerebrospinal fluid) and set aside for 30 minutes to make blood clots, the remaining samples can be tested directly, 1000Xg at room temperature and centrifuged for 10 minutes draw, supernatant (serum), immediately at -70 DEG C, refrigerator tested, TBI patients were divided into observation group two weight group, and 10 cases of control group (specimens for laboratory presentation of cerebrospinal fluid samples to exclude intracranial disease medicine Normal cerebrospinal fluid screened during disease was normal, and blood samples were collected by physical examination. All samples were detected by double antibody sandwich ELISA.
Results: the detection of MMP-9 reached the peak at 24 hours in TBI patients in the cerebrospinal fluid, at least for 72 hours, according to statistics, 24 hours and 72 hours had no significant difference, but the detection and within 120 hours there was a significant difference between the.TIMP-1 is within 120 hours of peak, the relative within seventy-second hours. Within twenty-fourth hours there was significant difference between.MMP-2 and TIMP-2 in cerebrospinal fluid in patients with TBI within 120 hours. No significant changes in the blood of each detection factor has no obvious change over time, MMP-2 TBI patients and control group MMP-9, TIMP-1, TIMP-2 and CST are both expression in the blood of the severe TBI were significantly different. A group of patients with MMP-9 in cerebrospinal fluid of patients with mild TBI group were significantly higher than that of MMP-2. However, the expression of TIMP-1 and TIMP-2 in cerebrospinal fluid in severe and mild group had no significant difference between MMP-2. MMP-9 in blood samples, TIMP-1 And the expression of TIMP-2 in severe and mild group had no significant difference between the group of patients with TBI. All of the injured after MMP-9 (120 hours) and the content of TIMP-1 (120 hours) were simple correlation analysis, results show that there is a linear correlation between negative correlation, correlation coefficient -0.593 (P0.01), MMP-2 with TIMP-2 in the detection and remain relevant.
Conclusion: the level of expression of MMP-9, MMP-2, TIMP-1 and TIMP-2 was detected and analyzed in different grades of TBI and normal brain tissue.
MMP-2 1.TBI patients and control group, MMP-9, TIMP-1 and TIMP-2 both in the blood or cerebrospinal fluid expression had significant differences in.MMP-9 reached the peak at 24 hours in TBI patients in the cerebrospinal fluid, at least lasts 72 hours. For patients with TBI, the main source of MMP-9 may be the nerve cells.
There was a simple correlation between 2.MMP-9 (120 hours) content and TIMP-1 (120 hours) expression in CSF. The difference between them is opposite..TIMP-1 may inhibit MMP-9..
The expression of 3.MMP-2 and TIMP-2 in the cerebrospinal fluid is negatively correlated with each detection point, and the change trend is contrary to.TIMP-2, which may inhibit MMP-2. specifically.
4.TIMP-1 may be an endogenous protective factor.
5.MMP-9 can be used as a predictor of clinical prognosis.

【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R651.15

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