Thrombin-induced autophagy: a potential role in intracerebral hemorrhage.
Thrombin-induced autophagy: a potential role in intracerebral hemorrhage.
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DOI：
10.1016/j.brainres.2011.09.062
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发表时间：
2011-11-18
期刊：
影响因子：
2.9
通讯作者：
Hua Y
中科院分区：
文献类型：
--
作者：
Hu S;Xi G;Jin H;He Y;Keep RF;Hua Y
脑出血（ICH）后脑内发生自噬，凝血酶参与ICH诱导的脑损伤和细胞死亡。在这项研究中，我们研究了凝血酶是否可以激活自噬（在体内和培养的星形胶质细胞）及其在ICH中的潜在作用。采用电子显微镜、轻链3（LC 3）从LC 3-I型转化为LC 3-II型、组织蛋白酶D蛋白质印迹和单丹酰尸胺（MDC）染色检测自噬空泡来检查自噬。3-甲基腺嘌呤（3-MA）被用作自噬抑制剂。在体内，我们发现尾状核内注射凝血酶增加了LC 3-I向LC 3-II的转化，组织蛋白酶D水平，以及同侧基底神经节中自噬空泡的形成。ICH诱导的LC 3-I向LC 3-II转化和组织蛋白酶D水平的上调被凝血酶抑制剂水蛭素降低。在培养的星形胶质细胞中，凝血酶增强了LC 3-I向LC 3-II的转化，并增加了MDC标记的自噬空泡。3-MA抑制凝血酶诱导的自噬空泡形成，并加剧凝血酶诱导的细胞死亡。这些结果表明凝血酶激活脑中的自噬，并且凝血酶在ICH诱导的自噬中起作用。
Autophagy occurs in the brain after intracerebral hemorrhage (ICH) and thrombin contributes to ICH-induced brain injury and cell death. In this study, we investigated whether thrombin may activate autophagy (in vivo and in cultured astrocytes) and its potential role in ICH. Autophagy was examined using electron microscopy, conversion of light chain 3(LC3) from the LC3-I form to LC3-II, cathepsin D Western blotting and monodansylcadaverine (MDC) staining to detect autophagic vacuoles. 3-Methyladenine (3-MA) was used as an autophagy inhibitor. In vivo, we found that intracaudate injection of thrombin increased conversion of LC3-I to LC3-II, cathepsin D levels, and formation of autophagic vacuoles in the ipsilateral basal ganglia. ICH-induced upregulation of LC3-I to LC3-II conversion and cathepsin D levels was reduced by a thrombin inhibitor, hirudin. In cultured astrocytes, thrombin enhanced the conversion of LC3-I to LC3-II and increased MDC-labeled autophagic vacuoles. 3-MA inhibited thrombin-induced autophagic vacuole formation and exacerbated thrombin-induced cell death. These results indicate that thrombin activates autophagy in the brain and that thrombin has a role in ICH-induced autophagy.