曾朝涛 张萌 周天恩 蒋龙元

DOI：10.3760/cma.j.issn.1671-0282.2014.08.013

基金項目：广东省自然科学基金（06021323）

作者单位：510120 广州，中山大学附属孙逸仙纪念医院急诊医学科

通信作者：蒋龙元，Email：jly@China-em.com

【摘要】目的 观察乌司他丁对脓毒症大鼠血脑屏障及脑细胞凋亡的影响。

访法 清洁级雄性SD大鼠52只，随机（随机数字法）分为6组。假手术（Sham）6 h组、24 h组，每组6只；脓毒症（CLP）及乌司他丁（UTI）6 h组、24 h组，每组10只。Sham组只开腹后关腹，CLP组及UTI组用盲肠结扎穿孔法（CLP）建立脓毒症模型，UTI组建模后1 h股静脉注射乌司他丁50 000 U/kg，其他组注射等容量生理盐水。各组在建模后6 h、24 h断脑取材，取材前进行神经功能缺损评分，取材后HE染色、称量脑干湿质量、伊文思蓝渗透法测定血脑屏障通透性，TUNEL免疫荧光检测脑细胞凋亡。应用SPSS 13.0软件统计学分析，多组比较采用方差分析，两两比较采用LSD-t检验。

结果 UTI组大鼠神经功能评分低于Sham组（P<0.05），高于CLP组（P<0.05）。光镜下CLP组大鼠神经细胞变性、坏死、水肿较UTI组明显。CLP 24 h组大鼠血脑屏障通透性较UTI 24 h组显著升高（P<0.05）。CLP组大鼠脑细胞凋亡数较UTI组显著增多（P<0.05）。

结论 乌司他丁通过减轻血脑屏障损伤，抑制脑细胞凋亡，对脓毒症大鼠脑组织起保护作用。

【关键词】脓毒症；脑损伤；盲肠结扎穿孔；乌司他丁；伊文思蓝；血脑屏障；免疫荧光；凋亡

Protective effect of ulinastatin on cerebral tissue in septic rats Zeng Chaotao， Zhang Meng， Zhou Tianen， Jiang Longyuan. Emergency Department of Sun Yat-Sen Memorial Hospital Afflicted to Sun Yat-Sen University， Guangzhou 510120， China.

Corresponding author： Jiang Longyuan， Email： jly@China-em.com

【Abstract】Objective To explore the effect of Ulinastatin on blood brain barrier （BBB） and apoptosis of neural cells in septic rats. Methods Fifty-two clean level male Sprague-Dawley rats were randomly（random number table） divided into six groups： Sham groups at 6 h and 24 h， each group with six rats. Sepsis groups（CLP） and Ulinastatin treated groups （UTI） at 6 h and 24 h， each group with ten rats. In CLP and UTI groups， cecal ligation and puncture （CLP） were performed to induce sepsis. Sham group was only opened and closed abdomen. Ulinastatin （50 000 U/kg） was administered via femoral vein 1 h after CLP. The same volume of saline instead of Ulinastatin was administered in Sham and CLP groups. The neurological status was assessed by Neurological Deficit Scale Scores （NDSS） at 6 h and 24 h after CLP. Then the brain was harvested for HE staining and weighing water content. The BBB permeability was assayed by Evans Blue dye extravasations. Apoptosis of neural cells were detected by TUNEL immune fluorescence. Statistical analysis was performed with SPSS version 13.0， ANOVA was used for multiple groups comparison and t-test for paired comparison. Results TheNeurological Deficit Scale Scores of UTI group was lower than Sham group（P<0.05） but higher than that of CLP group（P<0.05）. Swelling， degeneration and edema were observed in cerebral cortex and hippocampal neurons in CLP group through light microscope， and were more serious than those in UTI group. Compared with UTI 24 h group， BBB permeability of CLP 24 h group significantly rose（P<0.05）. The number of apoptosis of neural cells increased more in CLP group than it did in UTI group （P<0.05）. Conclusions Ulinastatin could protect the cerebral tissue in septic rats by alleviating the damage of BBB and reducing the apoptosis of neural cells.

【Key words】Sepsis； Cerebral injury； Cecal ligation and puncture； Ulinastatin； Evans blue； Blood brain barrier； Immune fluorescence； Apoptosis

脓毒症脑病是脓毒症的常见并发症，其严重程度与患者的病死率正相关^[1]。目前并无特殊针对脓毒症脑病的治疗措施。乌司他丁是从健康成年男性新鲜尿液中分离纯化出来的一种蛋白酶抑制剂，具有稳定溶酶体膜，抑制溶酶体酶释放，清除氧自由基及抑制炎症介质释放的作用，研究发现乌司他丁对脓毒症多器官有保护作用^[2-4]，但对脑组织影响的研究较少。本文试图探讨乌司他丁对脓毒症大鼠脑组织是否有保护作用及可能机制。……