脑室周围白质软化新生大鼠脑组织炎症因子变化  

A study on changes of brain tissue inflammatory cytokines in neonatal rats with periventricular leukomalacia

作　　者：李健[1] 欧阳颖[1] 黄科[2] 董红[1] 王琴[1] 

机构地区：[1]中山大学附属孙逸仙纪念医院新生儿科,广州510000 [2]中山大学附属孙逸仙纪念医院儿科血液专科,广州510000

出　　处：《中国新生儿科杂志》2017年第2期150-154,共5页Chinese Journal of Neonatology

基　　金：广东省自然科学基金自由申请项目（S2013010016308）

摘　　要：目的 探讨脑室周围白质软化（PVL）新生大鼠的炎症损伤情况.方法 选择3日龄SD大鼠80只,随机分为假手术组和PVL组各40只,每组又分为12、24、48、72 h4个亚组各10只.PVL组通过单侧颈总动脉结扎及术后吸入8％氧气＋92％氮气混合气2.5h制备缺血缺氧型PVL新生大鼠动物模型;假手术组仅游离左侧颈总动脉,不予结扎及缺氧.两组大鼠于4个时间点分别断头取脑组织,应用苏木精-伊红（HE）染色观察脑组织病理改变;实时荧光定量逆转录-聚合酶链反应（RT-qPCR）法检测肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β） mRNA表达水平;酶联免疫吸附试验（ELISA）法检测TNF-α及IL-1β3蛋白含量.结果 HE染色可见PVL组大鼠脑组织较假手术组明显充血、水肿,左侧脑室扩大,脑室周围白质结构稀疏、紊乱.PVL组TNF-α及IL-1β mRNA表达上调,均出现随时间推移先升高,24h达高峰,随后逐渐降低的趋势,且12、24、48 h两两比较,差异均有统计学意义（P ＜0.05）;72 h后TNF-α及IL-1β mRNA表达水平均下调,与同组48 h相比差异无统计学意义（P＞0.05）.PVL组各时间点TNF-α及IL-1β蛋白含量比较差异有统计学意义（P＜0.01）,且各时间点TNF-α蛋白含量均高于假手术组[12 h（189.2 ±20.4） pg/ml比（131.4 ±5.2） pg/ml、24h（213.8±16.7）pg/ml比（127.7±7.4） pg/ml、48 h（181.7±15.0）pg/ml比（126.3±6.0）pg/ml、72 h（159.6±25.3）pg/ml比（131.4±6.0） pg/ml,P＜0.01];IL-1β蛋白含量仅在24、48 h与假手术组相比差异有统计学意义[（121.8±30.0） pg/ml比（67.4±13.7） pg/ml、（83.3±15.7） pg/ml比（65.3±14.9）pg/ml,P＜0.05）].假手术组TNF-α及IL-1β蛋白含量各时间点比较,差异无统计学意义（P＞0.05）.结论 TNF-α与IL-1β共同参与缺血缺氧诱导的PVL,且具有先升高后降低的时间趋势,动态监测其变化有望成为PVL早期诊断、疗效观察、预后及评估的重要手段.Objective To study inflammatory cytokines changes in brain tissue of neonatal rats with periventricular leukomalacia （PVL）.Method A total of 80 neonatal SD rats （P3） were randomly assigned into 2 equal groups,sham-operated group and PVL group.Rats in each group were further assigned into four subgroups （12,24,48,72 h）,with 10 rats in each subgroup.The hypoxic-ischemic PVL modal were established following the procedure：first,isolation and ligation of left common carotid artery,and then exposed to 8％ O2 and 92％ N2 for 2.5 h.The sham-operated rats were processed with isolation of left common carotid artery only.Rats of the four subgroups were sacrificed at 12,24,48 h and 72 h respectively,then the brains were rapidly removed in corresponding time.Pathological changes of brain tissues were observed using HE stain.The mRNA expression levels of tumor necrosis factor α（TNF-α）and inter leukin-1β （IL-1β） were assessed using real-time quantitative PCR assays,the protein levels of TNF-α and IL-1β were detected using enzyme linked immunosorbent assay method.Result The brains tissues of rats in PVL group showed remarkably hyperemia and edema,with left ventricle enlargement.Periventricular white matter structure was disintegrated comparing with sham-operated group.The expression of TNF-α and IL-1 β mRNA in PVL group increased significantly,reaching peak by 24 h and then gradually decreased 72 h after the procedure.The mRNA levels of TNF-c and IL-1 β were significantly different between each two time points of 12,24 h and 48 h in PVL group （P ＜0.05）.However,there was no differences between 72 h and 48 h within PVL and sham-operated group group（P ＞0.05）.In PVL group,the protein expression trends of TNF-α and IL-1 β were similar to mRNA expression trends.Moreover,the protein levels were significantly different between each two time points of TNF-α and IL-1 β,respectively （P ＜ 0.01）.The protein expression levels of TNF-α were different at each time point between PVL group a

关 键 词：白质软化病 脑室周围 肿瘤坏死因子Α 白细胞介素1Β 大鼠 

分 类 号：R742[医药卫生—神经病学与精神病学]