乌司他丁对呼吸心跳骤停大鼠复苏早期脑组织谷氨酸代谢及GLT-1表达的影响

发布时间：2018-12-15 14:21

【摘要】：目的：窒息法制备大鼠心脏骤停模型和心肺复苏模型，探讨乌司他丁对海马组织病理变化、三磷酸腺苷及谷氨酸含量、谷氨酸转运体-1表达的影响。方法：成年SD雄性大鼠210只，随机数字表法分为假手术组（S组）：麻醉、气管切开及血管穿刺；乌司他丁复苏组（UTI组）：麻醉、气管切开及血管穿刺，窒息后常规复苏，ROSC后2min内和1.5h经股静脉推注乌司他丁(105U/kg)；生理盐水复苏组(NS组)：ROSC后相同时间点推注等剂量生理盐水。S组按气管切开、血管穿刺后分0.5h、1h、1.5h、2h、2.5h、4h、6h亚组；复苏组分为ROSC后0.5h、1h、1.5h、2h、2.5h、4h、6h亚组。光镜观察海马组织形态学改变，电镜观察海马组织超微结构变化；高效液相荧光测定大鼠海马组织Glu含量；高效液相法测定海马ATP含量；免疫组织化学法定位海马GLT-1的表达；Western blot测定GLT-1蛋白表达；RT-PCR测定GLT-1mRNA表达。结果：（1）NS组、UTI组大鼠海马组织主要病理改变包括海马神经元变性、坏死等，同时间点UTI组大鼠海马组织病理损伤轻于NS组。（2）与S组比较，NS组大鼠海马ATP含量在ROSC后七个时间点均降低，有统计学差异（P0.05，P0.01）；UTI组ATP含量在ROSC后0.5h、1h、1.5h、2h、2.5h较S组降低，有统计学差异（P0.05，P0.01）；UTI组与NS组比较，在ROSC后0.5h、4h、6h的ATP含量升高，有统计学差异（P0.01）。（3）与S组比较，NS组大鼠海马组织在ROSC后七个时间点Glu含量均明显升高，有统计学差异（P0.01）；UTI组较S组Glu在ROSC后1h、1.5h、2h、2.5h有所升高，有统计学差异（P0.05，P0.01）；与NS组比较，UTI组Glu浓度在ROSC后0.5h、2h、2.5h、4h、6h降低，有统计学差异（P0.05，P0.01）。（4）免疫组化定位海马组织GLT-1表达在神经胶质细胞胞膜上。（5）与S组比较，NS组GLT-1蛋白在4h、6h降低明显，，有统计学差异（P0.01）；UTI组与S组比较，在2.5h、4h、6h GLT-1表达升高，有统计学差异（P0.05，P0.01）；UTI组与NS组比较，GLT-1表达在2h、2.5h、4h、6h升高，有统计学差异（P0.05，P0.01）。（6）UTI组与S组比较，在ROSC后在4h、6h GLT-1mRNA表达的升高有统计学差异（P0.05）；UTI组与NS组比较，GLT-1mRNA自ROSC后2h起，其表达明显升高，这一状态一直维持到ROSC后6h，有统计学差异（P0.05）。结论：UTI可能通过改善能量代谢、上调GLT-1表达以及减少Glu含量，在一定程度上减轻了心肺复苏后海马组织病理损伤，对脑组织具有保护作用。
[Abstract]:Aim: to establish cardiac arrest model and cardiopulmonary resuscitation model in rats by asphyxia and to investigate the effects of ulinastatin on histopathological changes, adenosine triphosphate and glutamate content, and the expression of glutamate transporter 1 in hippocampus. Methods: 210 adult male SD rats were randomly divided into sham operation group (group S), anesthesia, tracheotomy and vascular puncture. Ulinastatin resuscitation group (UTI group): anesthesia, tracheotomy and vascular puncture, routine resuscitation after asphyxia, intravenous injection of ulinastatin (105U/kg) in 2min and 1.5 h after ROSC; Saline resuscitation group (NS group) was injected with the same dose of saline at the same time point after): ROSC. Group S was treated with tracheotomy. The resuscitation group was divided into subgroup of 0.5 h ~ 1 h ~ 1.5 h ~ 2 h ~ 4 h ~ 6 h after ROSC. Morphological changes of hippocampus were observed under light microscope, ultrastructure of hippocampus was observed by electron microscope, Glu content in hippocampus was measured by high performance liquid fluorescence (HPLC), ATP content in hippocampus was measured by HPLC. The expression of GLT-1 in hippocampus was determined by immunohistochemical method, and the expression of GLT-1 protein was detected by; Western blot and GLT-1mRNA by RT-PCR. Results: (1) the main pathological changes of hippocampal tissue in NS group and UTI group included degeneration and necrosis of hippocampal neurons. At the same time point, the pathological damage of hippocampus in UTI group was lighter than that in NS group. (2) compared with S group, the pathological changes of hippocampus in UTI group were lighter than those in S group. The content of ATP in hippocampus of NS group decreased at the seven time points after ROSC (P0.05-P0.01). The content of ATP in UTI group was significantly lower than that in S group at 0.5 h, 1.5 h and 2 h after ROSC (P0. 05 and P0. 01). Compared with NS group, the content of ATP in UTI group was significantly higher than that in NS group at 0.5 h after ROSC (P0.01). (3). Compared with S group, Glu content in hippocampus of NS group was significantly increased at seven time points after ROSC. There was statistical difference (P0.01). The Glu of UTI group was higher than that of S group at 1 hour, 1.5 h and 2 h after ROSC, and there was statistical difference (P0.05 卤P0.01). Compared with NS group, the concentration of Glu in UTI group was significantly lower than that in UTI group at 0.5 h, 2 h, 2.5 h, 4 h and 6 h after ROSC (P 0.05, P < 0.05). (5) compared with S group, the expression of GLT-1 protein in NS group was significantly lower than that in S group at 4 h or 6 h (P0.01). Compared with S group, the expression of GLT-1 in UTI group was significantly higher than that in S group at 2.5h, 4h and 6h (P0.05, P0.01). The expression of GLT-1 in UTI group was significantly higher than that in NS group (P0.05, P0.01). (6), and the expression of GLT-1 in UTI group was significantly higher than that in S group at 4 h after ROSC. The expression of GLT-1mRNA increased significantly at 6 h (P0.05). The expression of GLT-1mRNA in UTI group was significantly higher than that in NS group from 2 h after ROSC, which was maintained until 6 h after ROSC (P0.05). Conclusion: UTI may improve energy metabolism, up-regulate the expression of GLT-1 and decrease the content of Glu, which may alleviate the pathological injury of hippocampus after cardiopulmonary resuscitation to some extent, and has protective effect on brain tissue.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R614

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