溶酶体组织蛋白酶B增加自噬保护缺氧诱导的心脏微血管内皮细胞损伤  

Role of lysosomal cathepsin B in endothelial cell injury induced by hypoxia

作　　者：高路[1] 姚瑞[1] 李亚彭[1] 梁翠[1] 肖莉丽[1] 张彦周[1] GAO Lu;YAO Rui;LI Ya-peng;LIANG Cui;XIAO Li-li;ZHANG Yan-zhou(Dept of Cardiology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院心血管内科,河南郑州450052

出　　处：《中国药理学通报》2022年第1期53-60,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81970201);河南省优秀青年科学基金资助项目(No 212300410076)。

摘　　要：目的探讨CTSB是否参与缺氧诱导的心脏微血管内皮细胞损伤。方法采用缺氧诱导内皮细胞损伤模型;分离CTSB基因敲除小鼠心脏微血管内皮细胞,采用腺病毒过表达CTSB,采用巴弗洛霉素阻断细胞自噬;采用ELISA法检测炎症因子的释放水平;采用TUNEL染色法检测细胞凋亡数量;采用caspase-3试剂盒检测细胞caspase-3的活性;细胞感染LC3-GFP-mCherry双标病毒,检测细胞自噬流的水平。结果CTSB基因敲除可明显加重缺氧诱导的内皮细胞炎症、凋亡,增加细胞自噬。CTSB过表达可减轻缺氧诱导的内皮细胞炎症、凋亡,增加细胞自噬。但是巴弗洛霉素处理可明显抵消CTSB过表达对细胞炎症、凋亡的抑制作用和对细胞自噬的保护作用。结论 CTSB基因敲除加重缺氧诱导的内皮细胞炎症和凋亡,而CTSB基因过表达减轻缺氧诱导的内皮细胞损伤。CTSB通过维持内皮细胞正常的自噬降解发挥作用。Aim To investigate whether CTSB is involved in hypoxia-induced injury of cardiac microvascular endothelial cells. Methods A hypoxia-induced endothelial cell injury model was used. Cardiac microvascular endothelial cells were isolated from CTSB gene knockout mice. CTSB was overexpressed by adenovirus delivery system, and bafilomycin was used to block autophagy. ELISA was used to detect the release of inflammatory factors. Tunel staining was used to detect the number of cell apoptosis. caspase-3 kit was used to detect the activity of cell caspase-3. Cells were infected with LC3-GFP-mCherry double-labeled adenovirus to detect cell autophagy flow.Results CTSB gene knockout could significantly aggravate the inflammation and apoptosis of endothelial cells induced by hypoxia,and increased autophagy. Overexpression of CTSB reduced the inflammation and apoptosis of endothelial cells induced by hypoxia, and increased autophagy.But bafilomycin treatment could significantly offset the inhibitory effect of CTSB overexpression on cell inflam-mation and apoptosis and the protective effect on cell autophagy.Conclusions CTSB knockout aggravates inflammation and apoptosis induced by hypoxia in endothelial cells;while the overexpression of CTSB ameliorates endothelial cell injury induced by hypoxia.CTSB maintains normal autophagy degradation in endothelial cells. BAF blocks the protective effect of CTSB on endothelial cells by inhibiting autophagy degradation.

关 键 词：心脏缺氧损伤 内皮细胞损伤 溶酶体组织蛋白酶B 自噬 凋亡 炎症 

分 类 号：R-332[医药卫生] R332.11