摘要
目的探讨银杏内酯B对人脐静脉内皮细胞连接蛋白以及血管渗透性的影响。方法用银杏内酯B(0.2、0.4、0.6 g·L-1)孵育人脐静脉内皮细胞1 h,再用ox-LDL刺激细胞4 h,用Western blot及免疫荧光法检测JAM-A、Cx43的表达。用白细胞迁移实验观察ox-LDL损伤内皮细胞以及银杏内酯B对单核细胞向内皮细胞下迁移的影响。结果①ox-LDL刺激内皮细胞4 h,JAM-A表达增加了22%,Cx43表达增加了24%,银杏内酯B处理的细胞JAM-A、Cx43表达明显降低。免疫荧光检测获得了同样的结果。②PI3K特异性抑制剂LY294002同样抑制了ox-LDL刺激的JAM-A、Cx43表达,表明银杏内酯B降低JAM-A、Cx43表达与抑制Akt磷酸化相关。③白细胞迁移实验表明,银杏内酯B明显减少ox-LDL诱导的单核细胞向内皮细胞下迁移。结论银杏内酯B能抑制ox-LDL刺激的内皮细胞连接蛋白JAM-A、Cx43表达,从而改善血管渗透性,减少白细胞迁移,分子机制可能与抑制PI3K/Akt信号通路活化有关。
Aim To investigate the effect of ginkgolide B on junctional proteins in ox-LDL-stimulated human umbilical vein endothelial cells （HUVECs）. Methods After incubation with ginkgolide B （0.2,0.4,0.6 g . L-1） for 1 h, HUVECs were treated with ox-LDL （0. 1 g . L-1）for 4 h. The expressions of JAM-A and Cx43 were analyzed with Western blot and immunofluorescence. The effect of ginkgolide B on vascular permeability was analyzed by Transwell experiments. Results JAM-A and Cx43 expressions increased by 22% and 24% in ox-LDL-treated HUVECs, respectively. Whereas ginkgolide B significantly decreased JAM-A and Cx43 expressions. LY294002, a speeific inhibitor of PI3K, suppressed JAM-A and Cx43 expressions in ox-LDL-stimulated cells. Ginkgolide B potently reduced monocyte migration in ox-LDL-treated ceils. Conclusion Ginkgolide B significantly suppresses JAM-A and Cx43 expressions, and reduces monoeyte migration in ox-LDL-stimulated eells. This demonstrates that ginkgolide B can improve vaseular permeability. The mechanism might be associated with the inhibition of PI3K/Akt signaling pathway.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2014年第5期646-651,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81070231,81270379)
