缺氧缺血后脑水肿与水通道蛋白9表达及甘露醇作用相关机制研究  

Research on the expression of aquaporin 9 gene and efficacy of mannitol in neonatal rats with hypoxia-ischemic brain edema

作　　者：王芳会[1] 焦书真 樊书娟[1] 艾婷[1] 肖谧[1] 林晓洁[1] 刘俐[1] Wang Fanghui;Jiao Shuzhen;Fan Shujuan;Ai Ting;Xiao Mi;LinXiaojie;Liu Li(Department of Neonatology,First Affiliated Hospital of Xi＇an Jiaotong University,Xi＇an 710061,China)

机构地区：[1]西安交通大学第一附属医院新生儿科,710061 [2]西安医学院第一附属医院儿科

出　　处：《中华新生儿科杂志（中英文）》2018年第5期376-380,共5页Chinese Journal of Neonatology

基　　金：国家自然科学基金（81070539）

摘　　要：目的探讨新生大鼠缺氧缺血性脑损伤（hypoxia－ischemic brain damage，HIBD）后脑水肿与水通道蛋白（aquaporin，AQP）9的关系，以及甘露醇治疗脑水肿的机制。方法选择健康7日龄SD大鼠108只，随机分为HIBD组、假手术组、甘露醇组各36只。HIBD组和甘露醇组制备缺血缺氧模型，甘露醇组在HIBD后0、24、48 h腹腔注射甘露醇，动物处死前均经腹腔注射2%伊文斯蓝2 ml/kg，各组在HIBD后0、6、12、24、48、72 h处死并检测脑组织含水量，逆转录聚合酶链反应和免疫荧光染色测定AQP 9表达变化，用伊文斯蓝作为血脑屏障通透性指示剂测定血脑屏障通透性。结果HIBD组脑组织含水量从0 h起高于假手术组（P〈0.05），随时间延长持续增加，48 h达顶峰（89.3%±1.9%），随后渐消退；甘露醇组脑组织含水量在应用甘露醇1 h后开始下降，12 h达低谷（86.5%±0.6%），72 h上升（87.2%±1.7%），0～48 h均低于HIBD组（P〈0.05）。HIBD组伊文斯蓝含量0 h开始增加，48 h达峰值，各时间点均高于假手术组（P〈0.05）；甘露醇组伊文斯蓝含量各时间点均低于HIBD组，除0 h外，其余时间点差异均有统计学意义（P〈0.05）。HIBD组AQP 9 mRNA在0 h后下降，48 h达最低（0.09±0.07），0、6、72 h高于假手术组，12、24、48 h低于假手术组（P〈0.05）；除48 h外，甘露醇组各个时间点AQP 9 mRNA表达均高于HIBD组和假手术组（P〈0.05）；免疫荧光染色显示AQP 9广泛存在于软脑膜、室管膜等与水代谢密切相关的界面，缺血缺氧后早期AQP 9表达增加，随后下降，晚期再增加。结论AQP 9存在于脑组织各部位，通过多种途径参与脑水肿的形成和消退，在甘露醇减轻脑水肿中发挥作用。ObjectiveTo study the relationship between aquaporin 9（AQP 9） gene and brain edema in neonatal rats of hypoxic-ischemic brain damage （HIBD）and the therapeutic mechanism of mannitol.MethodHealthy and 7-day-old SD rats were randomly assigned into three groups： sham-operated group, HIBD group and mannitol group. Both HIBD and mannitol group were established on HIBD model. The mannitol group was given mannitol intraperitoneally at 0, 24, 48 h of HIBD. 2 ml/kg of 2% Evans blue （EB） were injected intraperitoneally before sacrifice. 0, 6, 12, 24, 48 and 72 h after HIBD, the outcomes were analyzed including the brain water content, the expression of AQP 9 mRNA measured using RT-PCR and immunofluorescence staining methods, and the permeability of blood-brain barrier （BBB） measured with EB.ResultIn HIBD group, the brain water content was higher comparing with sham-operated group at 0 h after HIBD（P〈0.05 ）, and gradually increased over time, reaching peak at 48 h （89.3%±1.9%）and then decreased. In mannitol group, brain water content started to decrease from 1 h after mannitol administration to the bottom at 12 h （86.5%±0.6%）, then increased to peak at 72 h （87.2%±1.7%）, and brain water content were decreased during 0~48 h comparing with HIBD group. HIBD group′s EB were higher than sham-operated group （P〈0.05）; Mannitol group′s EB were decreased comparing with HIBD group （except 0 h, P〈0.05）. AQP 9 mRNA expression in the HIBD group was decreased at 0 h, and reached the bottom at 48 h （0.09±0.07）. Comparing with sham-operated group, it was higher in the HIBD group at 0, 6, 72 h, and lower （P〈 0.05） at 12, 24, 48 h. Higher AQP 9 mRNA expression were detected in mannitol group than HIBD group and sham-operated group at each time point （with the exception of 48 h） （P〈 0.05）. AQP 9, which was closely related to water metabolism, were widely found in the pia mater and ependyma using immunofluorescence staining. After ischemia and hypoxia insult, an increas

关 键 词：缺氧缺血 脑 脑水肿 血脑屏障 水通道蛋白9 甘露醇 

分 类 号：R722.1[医药卫生—儿科]