凝血酶在脑出血后脑损伤过程中促进细胞自噬性死亡的研究  

The role of thrombin in intracerebral hemorrhage-induced autophagy

作　　者：胡枢坤[1] 吴靖宇[1] 黄峰平[1] 

机构地区：[1]复旦大学附属华山医院神经外科,上海200040

出　　处：《中华实验外科杂志》2014年第2期322-324,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 探讨凝血酶在脑出血后脑损伤过程中与细胞自噬性死亡的关系.方法 制作大鼠脑出血模型,干预组同时注射凝血酶抑制剂水蛭素.制作大鼠凝血酶脑损伤模型,对照组注射同体积生理盐水,应用免疫组织化学染色和Western blot检测自噬相关指标,电镜直接观察自噬细胞.结果 Western blot检测提示,脑出血大鼠注射水蛭素后的微管相关蛋白l轻链3（LC3）Ⅱ/LC3 Ⅰ为1.030±0.170,组织蛋白酶D（Cathepsin D）为（6.63±1.22）像素,低于单纯脑出血大鼠LC3Ⅱ/LC3 Ⅰ的1.410±0.080和Cathepsin D的9.12±0.72;大鼠注射凝血酶后的LC3Ⅱ/LC3 Ⅰ为0.730±0.070,Cathepsin D为10.34±2.22,高于对照组大鼠LC3Ⅱ/LC3 Ⅰ的0.402±0.104和Cathepsin D的4.94±1.11;电镜下可在凝血酶处理大鼠脑内检测到大量自噬细胞.结论 凝血酶可以促进大鼠脑出血后细胞自噬性死亡的激活.Objective Thrombin contributed to intracerebral hemorrhage （ICH） induced brain injury and cell death.Autophagy occurred after ICH.In this study,we are going to investigate the role of thrombin in ICH induced autophagy.Methods Male,adult Sprague-Dawley rats received infusion of autologus blood with or without hirudin （thrombin inhibitor） into the right caudate nucleus,the rats were sacrificed at day 7.Rats received an infusion of rat thrombin or vehicle into the right caudate nucleus and were killed 1,3 or 7 days later.Brains were used for Western blotting,immunohistochemistry and electron microscopy analyses.Results We found that microtubule-associated protein 1 light chain 3 （LC3） Ⅱ/LC3 Ⅰ（1.410 ± 0.080） and Cathepsin D （9.12 ± 0.72） levels in ipsilateral basal ganglia were upregulated after ICH and they were blocked by hirudin treatment （LC3 Ⅱ/LC3 Ⅰ 1.030 ±0.170 and Cathepsin D 6.63 ± 1.22）.LC3 Ⅱ/LC3 Ⅰ （0.730 ±0.070）,Cathepsin D （10.34 ±2.22） expression were increased in the ipsilateral basal ganglia after thrombin infusion comparing to saline injected （LC3 Ⅱ/LC3 Ⅰ 0.402 ±0.104,Cathepsin D 4.94 ± 1.11）,vacuole formation were observed under electron microscopy 3 days after thrombin injected.Conclusion Thrombin plays an important role in ICH induced autophagy.

关 键 词：脑出血 脑损伤 凝血酶 自噬 

分 类 号：R743.3[医药卫生—神经病学与精神病学]