蚤休薯蓣皂苷对内皮细胞的保护作用涉及TLR4/NF-κB途径  

Effect of rhizome saponins on TLR4 signal transduction pathways on ox-LDL damaged human endothelial cells

作　　者：高琳琳[1] 李福荣[2] 姚树桐[1] 桑慧[1] 司艳红[1] 焦鹏[3] 秦树存[3] 

机构地区：[1]泰山医学院病理生理学教研室,山东泰安271000 [2]泰山医学院药物化学教研室,山东泰安271000 [3]泰山医学院动脉粥样硬化研究所,山东泰安271000

出　　处：《中国药理学通报》2012年第11期1598-1602,共5页Chinese Pharmacological Bulletin

基　　金：山东省自然科学基金资助项目(No ZR2011HL014);山东省高等学校科技计划项目(No J10LF18)

摘　　要：目的观察蚤休薯蓣皂苷(Rhizome Dioscin,RD)对氧化性低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HU-VEC)损伤对细胞生长、炎症反应Toll样受体4(TLR4)和细胞核因子κB(nuclear factor kappa B,NF-κB)表达的影响,探讨RD抗动脉粥样硬化(AS)的作用机制。方法体外培养HUVEC,RD 3个浓度预处理,加入ox-LDL,运用流式细胞术PI染色检测细胞周期,激光共聚焦显微镜JC-1染色检测线粒体电位(ΔΨm),Western blot检测TLR4和NF-κB蛋白的表达。结果与正常对照组相比,经ox-LDL损伤后HU-VEC细胞周期阻滞在G0/G1期,表现为G0/G1期细胞比例明显升高(P<0.01),S期细胞比例明显下降(P<0.01);细胞ΔΨm明显下降;TLR4和NF-κB的表达与正常组比较均升高。RD预处理能抑制ox-LDL诱导的细胞生长停滞,使细胞周期S期细胞比例增加(P<0.01),ΔΨm上升,TLR4和NF-κB的表达与损伤组相比均下降。结论抑制TLR4/NF-κB信号转导途径可能是RD抗内皮细胞氧化损伤、阻止经线粒体途径的细胞生长停滞的可能机制之一。Aim To observe the effect of Rhizome Di-oscin（RD）on cell growth,Toll like receptor 4（TLR4） and the nuclear factor kappa B（NF-κB）signal transduction pathways on human umbilical vein endothelial cells HUVEC damaged by oxidizing low density lipoprotein（ox-LDL）,and to discuss the anti-atherosclerosis（AS）mechanism of the RD.Methods Three concentration RD pretreated,HUVEC cultured in vitro,flow cytometry was used to detect cell cycle.Laser scanning confocal microscope JC-1 dyeing to detect mitochondria membrane potential（ΔΨm）and western blot to test TLR4 and the NF-κB protein expression. Results Cell cycle in ox-LDL injured group was blocked in G 0 / G 1 phase and cell number of G 0 / G 1 phase was increased（P 0.01）but cell number of S phase was decreased（P 0.01）.The mitochondrial membrane potential was obviously decreased,expression of TLR4 and the NF-κB in injured group were increased compared with normal control.RD group could block cell growth stagnation induced by ox-LDL,make the cell number of G 0 / G 1 phase decrease and S phase increase in the meantime（P 0.01）,ΔΨm rose. TLR4 and NF-κB expression were decreased compared with injured control.Conclusion RD can inhibit TLR4 / NF-κB signal transduction pathway on HUVEC, restrain the mitochondrial oxidative damage by the way of cell growth stagnation and this may be one of the possible mechanisms of anti-AS of RD.

关 键 词：蚤休薯蓣皂苷 氧化性低密度脂蛋白 内皮细胞 细胞周期 线粒体膜电位 Toll样受体4 细胞核因子ΚB 

分 类 号：R284.1[医药卫生—中药学] R322.123[医药卫生—中医学]