【摘要】 目的:分析光化学法诱导小鼠局灶性脑梗死模型脑组织肿瘤坏死因子α表达和髓过氧化物酶活性的变化规律及其相关性。方法:实验于2004-07/2005-07在泰山医学院脑微循环实验室完成。选用雄性KM小鼠60只,随机分为3组,正常对照组(n=6)、假手术组(n=6)、局灶性脑梗死组(n=48),局灶性脑梗死组按光照后时间分为30min,1,3,6,12,24,48,72h8个亚组,每亚组6只。采用光化学法制作小鼠局灶性脑梗死模型,假手术组除不作光照外其他操作同局灶性脑梗死组。采用免疫组化、酶联免疫吸附法检测梗死侧皮质肿瘤坏死因子α的表达,比色法检测梗死侧皮质髓过氧化物酶的活性,髓过氧化物酶活性可以反映组织中中性粒细胞的浸润程度,活性越高浸润越严重。结果:60只小鼠均进入结果分析。①脑梗死后肿瘤坏死因子α阳性细胞主要为神经元及胶质细胞,分布于梗死区及其周围。假手术组皮质肿瘤坏死因子α的表达量为(615.7±16.1)ng/L,局灶性脑梗死组于光照后3h开始明显升高(792.2±17.8)ng/L,6h达高峰(921.9±23.9)ng/L,12h开始下降(848.0±30.6)ng/L,72h恢复至正常水平(625.3±14.3)ng/L。②假手术组皮质髓过氧化物酶的活性为(7.151±0.433)nkat/g,局灶性脑梗死组于光照后3h开始明显升高(10.469±0.600)nkat/g,12h达高峰(15.486±0.650)nkat/g,24h开始下降(11.052±0.617)nkat/g,72h恢复至正常水平(7.418±0.617)nkat/g。③髓过氧化物酶活性变化稍滞后于肿瘤坏死因子α的表达,相关性分析示二者呈正相关(r=0.953,P<0.01)。结论:在光化学法诱导小鼠局灶性脑梗死急性期,肿瘤坏死因子α表达水平与髓过氧化物酶活性呈正相关,提示肿瘤坏死因子α可诱导炎性细胞侵入缺血脑组织,参与脑缺血早期的炎症反应过程。更多还原

【Abstract】 AIM: To analyze the expression of tumor necrosis factor alpha (TNF-α),the changes of activity of myeloperoxidase and the correlation between them in focal cerebral infarction based on the principle of photochemical initiation. METHODS: The experiment was performed at the Laboratory of Cerebral Microcirculatin,Taishan Medical University from July 2004 to July 2005. Totally 60 male KM mice were selected and divided randomly into normal control group (n=6),sham operation group (n=6) and focal cerebral infarction group (n=48). Focal cerebral infarction group were divided into 8 subgroups,including 30 minutes,1,3,6,12,24,48 and 72 hours subgroups,according to the time after lighting with 6 mice in each subgroup. The mice focal cerebral infarction models were made by the principle of photochemical initiation. The sham operation group was not lighted and the other disposals were the same to the focal cerebral infarction group. The immunohistochemistrical method and enzyme-linked immunoadsorbent assay were used to detect the expression of TNF-α in the infarction cortex. The activity of myeloperoxidase in the infarction cortex was tested by chromatometry. The activity of myeloperoxidase could reflex the infiltrating degree of neutrophile granulocytes,and the higher activity represented more severe infiltration. RESULTS: All the 60 mice were involved in the result analysis. ①After cerebral infarction,the positive cells of TNF-αwere distributed in neurons and glial cells and most of them were in or around the infarction region. The expression of TNF-α in the sham operation group were (615.7±16.1) ng/L,and it increased significantly from the 3rd hour after lighting in the local cerebral infarction group (792.2±17.8) ng/L,peaked at the 6th hour (921.9±23.9) ng/L,decreased after the 12th hour (848.0±30.6) ng/L,and then declined to normal level at the 72nd hour (625.3±14.3) ng/L. ②The activity of myeloperoxidase in the infarction cortex was (7.151±0.433) nkat/g,and increased significantly from the 3rd hour after lighting (10.469 ±0.600) nkat/g,peaked at the 12th hour (15.486±0.650) nkat/g,decreased to (11.052±0.617) nkat/g after the 24th hour,and then declined to normal level at the 72nd hour (7.418±0.617) nkat/g . ③The changes of the activity of myeloperoxidase lagged lightly behind the expression of TNF-α in the infarction cortex. Both of them had positive correlation showed by correlation analysis (r=0.953,P < 0.01). CONCLUSION: In focal cerebral infarction based on the principle of photochemical initiation in mice,the expression of TNF-α has positive correlation with the activity of myeloperoxidase,which indicates that the TNF-α can induce leucocytes infiltrate to the cerebral ischemia tissue and participate in inflammatory course during early cerebral ischemia.更多还原