摘要
目的探讨IL-1β对血管内皮细胞损伤机制。方法以EA.hy926血管内皮细胞为研究对象,利用不同浓度IL-1β刺激24 h,采用Western blot方法检测IL-1β对β1整合素、Bcl-2、Bax、Caspase-3及N-乙酞氨基葡萄糖转移-V(GnT-V)表达的影响;通过Lectin blot方法显示IL-1β对GnT-V所修饰的所有糖蛋白量的影响;免疫共沉淀方法检测IL-1β对β1整合素糖基化修饰量的变化;流式细胞术检测细胞凋亡率的变化。结果 IL-1β可从蛋白翻译水平抑制β1整合素的表达,同时可以下调Bcl-2/Bax的比例,上调Caspase-3的表达。初步揭示IL-1β诱导血管内皮细胞凋亡可能是通过β1整合素N-连接糖基化作用来完成的。结论 IL-1β可通过调控EA.hy926细胞β1整合素的糖基化修饰,增加细胞凋亡率,损伤血管内皮细胞。
Aim To investigate the effect of IL-1β on damaging human vascular endothelial cells（EA.hy926）. Methods This study mainly takes the EA.hy926 vascular endothelial cells（VEC） as its object.After treated with different concentrations of IL-1β for 24h,the effect of IL-1β on the levels of β1-integrin,Bcl-2,Bax,Caspase-3 and GnT-V was detected by Western blot.The effects of IL-1β on GnT-V product in the whole glycoprotein lysate were examined by lectin blot.Immunoprecipitation was used to test GnT-V product（L-PHA reactivity） on β1-integrin glycoprotein induced by IL-1β.Cell apoptotic rate was detected by flow cytometry.Results IL-1β promote β1-integrin expression at protein translation level.Moreover,it could downregulate the Bcl-2/Bax ratio,and increase Caspase-3 expression.Apoptosis of vascular endothelial cells induced by IL-1β may be mediated by N-glycosylation modification of β1-integrin.Conclusion IL-1β induces EA.hy926 apoptosis via N-glycosylation modification on β1-integrin protein,which may cause dysfunction of vascular endothelial cells.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2012年第7期921-925,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81070222)
