柚皮苷抑制高糖诱导的脐静脉内皮细胞与单核细胞的粘附作用  

Naringin inhibits monocyte adhesion to high glucose-induced human umbilical vein endothelial cells

作　　者：熊莺[1,2] 王广发[1] 张俊艳[1] 吴少瑜[1] 徐伟[1] 张嘉杰[1] 吴曙光[1] 饶进军[1] 

机构地区：[1]南方医科大学药学院,广东广州510515 [2]中山大学附属第一医院,广东广州5100080

出　　处：《南方医科大学学报》2010年第2期321-325,共5页Journal of Southern Medical University

基　　金：广州市粤港关键领域重点突破项目(2006Z1-E6021);广东省自然科学基金(9151063201000008)

摘　　要：目的研究柚皮苷对高糖诱导的脐静脉内皮细胞(HUVECs)与单核细胞的粘附的抑制作用及机制。方法分离培养HUVECs后,采用不同剂量的柚皮苷预处理HUVECs,然后高糖诱导HUVECs;加入荧光染料标记单核细胞系THP-1细胞与HUVECs共同孵育,荧光显微镜下观察HUVECs与单核细胞的粘附作用并拍照,比较粘附密度;提取总蛋白后,Western blotting检测处理后HUVECs粘附分子的表达;荧光染料结合法检测处理后HUVECs活性氧的产生;提取处理的脐静脉内皮细胞核蛋白后,Western blotting检测核因子-κB在核内的量。结果高糖诱导HUVECs与单核细胞的粘附,柚皮苷可显著抑制高糖诱导的HUVECs与单核细胞的粘附。柚皮苷抑制高糖诱导的HUVECs细胞间粘附分子和血管内皮细胞粘附分子的表达,柚皮苷抑制高糖诱导的HUVECs活性氧产生,柚皮苷抑制高糖诱导的HUVECs核因子-κBp65活化。结论柚皮苷可能通过抑制高糖诱导的HUVECs活性氧产生,从而抑制核因子-κB的活化,进一步抑制细胞间粘附分子和血管内皮细胞粘附分子的表达,影响HUVECs与THP-1细胞的粘附,从而抑制高糖诱导的血管炎症。Objective To investigate the inhibitory effect of naringin on monocyte adhesion to high glucose-induced human umbilical vein endothelial cells（HUVECs）.Methods Cultured HUVECs isolated from human umbilical cords were pretreated with or without naringin and induced with high glucose（33 mmol/L） for 48 h.Human monocyte THP-1 cells,after labeling with BCECF-AM,were co-cultured with the HUVECs for 30 min.The labeled THP-1 cells adhering to HUVECs were observed under fluoroscence microscope,and the inhibitory effect of naringin on the cell adhesion was evaluated by measuring the adhering cell density.Western blot analysis was used to detect the expressions of the adhesion molecules in the HUVECs,and reactive oxygen species（ROS） production in the HUVECs was measured using an oxidation-sensitive fluorescent probe（DCFH-DA）.The nuclear extracts of the HUVECs were prepared to examine the expression of nuclear factor-kappa B（NF-κB） in the cell nuclei by Western blotting.Results HUVECs in high-glucose culture showed increased adhesion to THP-1 cells and enhanced expressions of the cell adhesion molecules,which were significantly attenuated by pretreatment with naringin（10-50 μg/ml）.High glucose induced DCF-sensitive intracellular ROS production in the HUVECs,and this effect was inhibited by naringin pretreatment of the cells.Naringin also suppressed high glucose-induced increment of NF-κB expression in the cell nuclei of HUVECs.Conclusion Naringin can suppress high glucose-induced vascular inflammation possibly by inhibiting ROS production and NF-κB activation in HUVECs.

关 键 词：柚皮苷 人脐静脉内皮细胞 单核细胞 粘附分子 活性氧 转录因子 糖尿病并发症 

分 类 号：R285.5[医药卫生—中药学]