盐酸青藤碱诱导EA.hy926细胞自噬及其在抗炎中的作用  

Anti-inflammation activity of sinomenine-induced autophagy in EA.hy926 cells

作　　者：杨亭[1] 倪振洪[1] 龚薇[1] 何凤田[1] 

机构地区：[1]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038

出　　处：《第三军医大学学报》2013年第11期1084-1087,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81001677);中国博士后科学基金(2011M500153);重庆市自然科学基金(2010BB5171);第三军医大学青年人才创新基金(2010XQN03)~~

摘　　要：目的探讨中药单体提取物盐酸青藤碱诱导人内皮细胞EA.hy926自噬的机制及其在抗炎中发挥的作用。方法Western blot检测分别以终浓度为50、100μg/mL的盐酸青藤碱处理12h后的EA.hy926细胞中自噬相关蛋白LC3Ⅱ、ERK2、磷酸化ERK2及炎症细胞因子HMGB1表达变化情况;荧光显微镜观察吖啶橙染色的经盐酸青藤碱诱导后EA.hy926细胞酸性小体变化情况。结果经终浓度为50、100μg/mL的盐酸青藤碱处理EA.hy926细胞后,与对照组比较,100μg/mL的盐酸青藤碱可使自噬相关蛋白LC3Ⅱ表达(0.67±0.05)及ERK2的磷酸化水平(1.08±0.05)上调(P<0.05),ERK抑制剂U0126可使LC3Ⅱ表达下调(P<0.05)及EA.hy926细胞中酸性小体减少;盐酸青藤碱可抑制EA.hy926中LPS诱导的HMGB1表达,自噬抑制剂氯喹可逆转该细胞中盐酸青藤碱对LPS诱导HMGB1表达的抑制作用(P<0.05)。结论盐酸青藤碱可通过ERK通路诱导EA.hy926细胞自噬,该自噬过程是盐酸青藤碱下调炎症细胞因子HMGB1进而发挥抗炎活性的机制之一。Objective To investigate the underlying mechanism of sinomenine, a traditional Chinese herb extract monomer, in inducing autophagy and its anti-inflammation activity in EA.hy926 cells. Methods Western blotting was taken to detect the expression of autophagic markers LC3Ⅱ, ERK2, phosphorylation ERK2 and inflammatory cytokine HMGB1 in the EA.hy926 cells after the treatment of sinomenine at 50 or 100 μg/mL for 12 h. Acridine orange staining was used to observe the formation of acidic vesicular organelles in the cells by fluorescence microscopy. Results Western blotting revealed that the expression of LC3Ⅱ （0.67±0.05）and phosphorylation ERK2 （1.08±0.05）were remarkably increased after the treatment of sinomenine at 100 μg/mL （P〈0.05）. Autophagic vesicles were induced by sinomenine,while both LC3Ⅱ and autophagic vesicles were partially decreased by ERK-specific inhibitor U0126 （P〈0.05）. LPS-induced HMGB1 expression could be reduced by sinomenine,which was partially reversed by autophagy inhibitor CQ （P〈0.05）. Conclusion It may be one of the mechanisms of anti-inflammation activity of sinomenine that can reduce HMGB1 by promoting autophagy through the ERK pathway in EA.hy926 cells.

关 键 词：盐酸青藤碱 自噬 HMGB1 ERK 抗炎 

分 类 号：R282.71[医药卫生—中药学] R285.5[医药卫生—中医学]