摘要
目的探讨内毒素(LPS)致脑损伤大鼠模型额叶皮质、海马组织、脑脊液(CSF)及血浆中脑红蛋白(Ngb)的表达变化及其意义。方法SD大鼠70只,随机分为对照组(n=10)和LPS干预组(6个时间点,每个时间点10只)。LPS干预组向大鼠脑池内注射0.1mg/kg LPS0.2ml,对照组注入等量生理盐水。注射LPS后3、6、12、24、48、72h收集血浆、CSF,并处死大鼠留取额叶皮质和海马组织。应用ELISA、Western blotting和免疫组织化学SP法检测Ngb的表达变化;应用干湿重法检测鼠脑含水量;应用投射电子显微镜检测大鼠额叶皮质和海马组织神经元细胞器的肿胀程度。结果注射LPS后,鼠脑含水量明显高于对照组(P<0.01),48h达峰值,为83.33%±1.90%。电镜照片显示,注射LPS后12h额叶皮质和海马组织中神经元发生明显肿胀,48h后达峰值。ELISA结果显示,LPS干预组额叶皮质、海马组织、CSF和血浆中Ngb含量,均显著高于对照组(P<0.01),48h达峰值,分别为35.37±3.91、29.36±3.99、22.71±3.12、14.39±2.77ng/ml。Western blotting结果显示LPS干预各组Ngb蛋白表达量均显著高于对照组,以3h组表达量最低,48h组表达量最高。免疫组织化学检测结果显示,Ngb阳性物质主要位于神经元细胞质,呈黄褐色。结论在LPS所致的脑损伤中Ngb表达明显上调,此现象可能与神经元内源性保护机制有关。
Objective To study the expression of neuroglobulin （Ngb） in cortex of frontal lobe, hippocampus, cerebrospinal fluid （CSF） and plasma of rats during the process of brain edema induced by endotoxin （LPS）. Methods Seventy SD rats were randomly divided into the control group （n=10） and the LPS groups （10 each at six time points）. An intraventricular injection of LPS （0.1mg/kg, 0. 2ml） was given to rats in LPS groups, while the same amount of normal saline was given in control group. The plasma, CSF, hippocampus and the cortex of frontal lobe of rats were collected 3, 6, 12, 24, 48 and 72 hours after the injection. Ngb was detected by ELISA, Western blotting and immunohistoehemical （IHC） method. The brain water content was measured by wet/dry method. The swelling of organelles in hippoeampus and the cortex of frontal lobe were observed by transmission electron microscopy （TEM）. Remits The brain water content was significantly higher in LPS groups than in control group （P〈0. 01）, and peaked （83. 33%±1. 90%） at 48h after LIPS injection. TEM showed that the neurons in hippocampus and the cortex of frontal lobe swelled remarkably 12h after LPS injection, and swelling was most marked at 48h. ELISA showed that the Ngb concentrations in the cortex of frontal lobe, hippocampus, CSF and plasma were all significantly higher in I.PS groups （35. 37, 29. 36, 22. 71, 14. 39ng/ml, respectively） than in control group （P〈 0.01）, and peaked at 48h. Western blotting showed that the expression of Ngb was significantly higher in LPS groups than in control group, and reached the highest level at 48h, and the lowest level at 3b. IHC showed that Ngb-positive cells distributed mainly in the cytoplasm of pyramidal neurons, and were yellowbrown in color. Conclusions Ngb is over-expressed markedly during the process of brain edema induced by LPS in rat, and it might be one of the endogenous nerve protective mechanisms.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2009年第9期1140-1143,共4页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金资助项目(30270437
30771109)
教育部春晖计划资助项目(2003-26)
