基质金属蛋白酶-9在结核性脑膜炎小鼠中的表达特点及意义

发布时间：2018-03-03 05:20

本文选题：结核性脑膜炎　切入点：基质金属蛋白酶-9　出处：《南昌大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的： 1、分析基质金属蛋白酶-9(MMP-9)在小鼠结核性脑膜炎病理过程中的表达特点及意义。 2、探讨单核细胞-星形胶质细胞网络对MMP-9表达的作用。方法： 1、从32只C57BL/6小鼠中随机抽取16只,向侧脑室注射10ul含（1.271±0.111）E+6cfu的H37RV菌悬液作为实验组，另16只用同样方法注射10ul生理盐水作为对照组,30天后处死所有小鼠。 2、分别从两组中随机抽取4只小鼠，取一侧脑组织进行结核菌培养，另一侧进行病理HE染色和免疫荧光共聚焦染色。 3、分别从两组中随机抽取4只小鼠，进行脑组织明胶酶谱检测。 4、分别从两组中随机抽取4只小鼠，进行血脑屏障通透性检测。 5、分别从两组中随机抽取4只小鼠，进行脑组织含水量检测。 6、采用两样本t检验比较两组差异，以P0.05为差异有统计学意义。结果： 1、实验组小鼠脑组织匀浆液培养出结核菌落(4.900±1.407)E+4cfu/ml，病理HE染色表现出蛛网膜下腔、脑室扩张，及大量炎症细胞浸润等典型脑膜炎病变，，对照组小鼠脑组织匀浆液均未培养出结核菌落，病理HE染色未见明显异常。 2、结核性脑膜炎小鼠脑组织MMP-9活性明显增强,实验组明胶酶谱法MMP-9条带累积光密度值为47821.25±19931.84,对照组为10081.50±3544.23(t=3.728,P=0.010)，差异有统计学意义。 3、结核性脑膜炎小鼠血脑屏障通透性增加，实验组脑组织伊文斯兰含量为11.8239±3.5954ug/g，对照组为4.6823±3.4009ug/g(t=2.887，P=0.028)，差异有统计学意义。 4、结核性脑膜炎小鼠脑组织含水量增加，实验组小鼠脑组织含水量比重为0.8492±0.0348，对照组为0.7752±0.0368(t=2.925，P=0.026)，差异有统计学意义。 5、免疫荧光染色显示实验组小鼠脑组织中MMP-9、GFAP和OX-42表达量均明显增多，且MMP-9与GFAP和OX-42均分别大量重合，表明结核性脑膜炎小鼠小胶质细胞、星形胶质细胞活化并分泌大量MMP-9。结论： 1、MMP-9在结核性脑膜炎小鼠中活性明显增强。 2、MMP-9通过分解细胞外基质破坏血脑屏障，促进白细胞、炎性介质进入脑组织参与结核性脑膜炎的病理过程。 3、小胶质细胞-星形胶质细胞网络参与调控MMP-9的分泌。
[Abstract]:Objective:. 1. To analyze the expression and significance of matrix metalloproteinase-9 (MMP-9) in the pathological process of murine tuberculous meningitis. 2. To investigate the effect of monocyte-astrocyte network on MMP-9 expression. Methods:. 1, 16 of 32 C57BL / 6 mice were randomly selected, and 10ul H37RV suspension containing 1.271 卤0.111Cfu was injected into the lateral ventricle as the experimental group. The other 16 mice were sacrificed after 30 days of injection of 10 ul normal saline as the control group with the same method. 2, 4 mice were randomly selected from the two groups, one side of brain tissue was cultured with tuberculous bacteria, the other side was stained with HE and immunofluorescence confocal staining. 3. Four mice were randomly selected from the two groups to detect the gelatinase spectrum of brain tissue. 4. Four mice were randomly selected from the two groups to detect the blood-brain barrier permeability. 5. Four mice were randomly selected from the two groups to measure the water content of brain tissue. 6, two samples of t test were used to compare the difference between the two groups, with P0.05 as the difference was statistically significant. Results:. 1. The mice in the experimental group were cultured in the homogenate of brain tissue of the experimental group. The mycobacterium tuberculosis was 4.900 卤1.407E 4cfu-ml. the pathological HE staining showed typical meningitis lesions such as subarachnoid space, ventricular dilatation, and a large number of inflammatory cell infiltration. In the control group, no tuberculous bacteria were found in the homogenate of brain tissue of the control group, and no obvious abnormality was found in the pathological HE staining. (2) the activity of MMP-9 in the brain tissue of mice with tuberculous meningitis was significantly increased. The cumulative optical density of MMP-9 bands in the experimental group was 47821.25 卤19931.84, and that in the control group was 10081.50 卤3544.23t0. 728P0.0100.The difference was statistically significant. 3, the permeability of blood-brain barrier increased in mice with tuberculous meningitis. The content of Ivanselam in brain tissue of experimental group was 11.8239 卤3.5954ugr / g, and that of control group was 4.6823 卤3.4009ugP / g / g (2.887g / g), the difference was statistically significant. (4) the water content of brain tissue increased in mice with tuberculous meningitis. The proportion of water content in brain tissue of experimental group was 0.8492 卤0.0348, and that of control group was 0.7752 卤0.0368. The difference was statistically significant. 5. Immunofluorescence staining showed that the expression of MMP-9 GFAP and OX-42 in the brain tissue of the experimental group was significantly increased, and that the expression of MMP-9, GFAP and OX-42 coincided with each other, indicating that the microglia and astrocytes of the mice with tuberculous meningitis were activated and secreted a large amount of MMP-9. Conclusion:. 1 the activity of MMP-9 was significantly increased in tuberculous meningitis mice. 2MMP-9 destroys the blood-brain barrier by breaking down the extracellular matrix and promotes the entry of leukocytes and inflammatory mediators. Brain tissue is involved in the pathological process of tuberculous meningitis. 3. Microglial-astrocyte network is involved in the regulation of MMP-9 secretion.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R529.3

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