中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
2期
322-324
,共3页
脑出血%脑损伤%凝血酶%自噬
腦齣血%腦損傷%凝血酶%自噬
뇌출혈%뇌손상%응혈매%자서
Intracerebral hemorrhage%Brain injury%Thrombin%Autophagy
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.
目的 探討凝血酶在腦齣血後腦損傷過程中與細胞自噬性死亡的關繫.方法 製作大鼠腦齣血模型,榦預組同時註射凝血酶抑製劑水蛭素.製作大鼠凝血酶腦損傷模型,對照組註射同體積生理鹽水,應用免疫組織化學染色和Western blot檢測自噬相關指標,電鏡直接觀察自噬細胞.結果 Western blot檢測提示,腦齣血大鼠註射水蛭素後的微管相關蛋白l輕鏈3(LC3)Ⅱ/LC3 Ⅰ為1.030±0.170,組織蛋白酶D(Cathepsin D)為(6.63±1.22)像素,低于單純腦齣血大鼠LC3Ⅱ/LC3 Ⅰ的1.410±0.080和Cathepsin D的9.12±0.72;大鼠註射凝血酶後的LC3Ⅱ/LC3 Ⅰ為0.730±0.070,Cathepsin D為10.34±2.22,高于對照組大鼠LC3Ⅱ/LC3 Ⅰ的0.402±0.104和Cathepsin D的4.94±1.11;電鏡下可在凝血酶處理大鼠腦內檢測到大量自噬細胞.結論 凝血酶可以促進大鼠腦齣血後細胞自噬性死亡的激活.
목적 탐토응혈매재뇌출혈후뇌손상과정중여세포자서성사망적관계.방법 제작대서뇌출혈모형,간예조동시주사응혈매억제제수질소.제작대서응혈매뇌손상모형,대조조주사동체적생리염수,응용면역조직화학염색화Western blot검측자서상관지표,전경직접관찰자서세포.결과 Western blot검측제시,뇌출혈대서주사수질소후적미관상관단백l경련3(LC3)Ⅱ/LC3 Ⅰ위1.030±0.170,조직단백매D(Cathepsin D)위(6.63±1.22)상소,저우단순뇌출혈대서LC3Ⅱ/LC3 Ⅰ적1.410±0.080화Cathepsin D적9.12±0.72;대서주사응혈매후적LC3Ⅱ/LC3 Ⅰ위0.730±0.070,Cathepsin D위10.34±2.22,고우대조조대서LC3Ⅱ/LC3 Ⅰ적0.402±0.104화Cathepsin D적4.94±1.11;전경하가재응혈매처리대서뇌내검측도대량자서세포.결론 응혈매가이촉진대서뇌출혈후세포자서성사망적격활.
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.
