瓜蒌皮提取物对高糖诱导人脐静脉内皮细胞凋亡的影响及机制研究  

Effects of extractive pericarpium trichosanthes on apoptosis of HUVECs induced by high glucose

作　　者：刘思妤[1] 谷彬[1] 卢新华[1] 王俊杰[1] 谭斌[1] 

机构地区：[1]湘南学院基础医学部,湖南郴州423000

出　　处：《中国药理学通报》2015年第7期988-993,共6页Chinese Pharmacological Bulletin

基　　金：湖南省中医药管理局重点基金项目(No 201227);湖南省卫生厅科研基金项目(No B2011-073);湖南省教育厅十二.五重点建设学科基金项目资助(No湘教发[2011]76号);湖南省心脑血管天然药物研究重点实验室基金项目资助(No湘教通[2008]246号)

摘　　要：目的探讨瓜蒌皮提取物(extractive pericarpium trichosanthes,EPT)对体外高糖诱导人脐静脉内皮细胞凋亡的抑制作用及其机制。方法体外培养人脐静脉内皮细胞株(HUVECs),观察不同浓度瓜蒌皮提取物干预高糖处理的HUVECs凋亡的变化。MTT法检测HUVECs的存活率、采用Hoechst染色法检测细胞凋亡、采用比色法测定细胞内Caspase-3的活性、Western blot法以及免疫荧光染色法检测核因子-κB(nuclear factor kappa B,NF-κB)的蛋白表达和p65核移位情况。结果 30 mmol·L-1葡萄糖处理内皮细胞48h能引起细胞活性明显降低,细胞凋亡率及caspase-3的活性明显增加,细胞中NF-κB蛋白表达升高及p65核移位增加。用瓜蒌皮提取物(12.5、25、50 mg·L-1)预处理细胞1 h,可以增加细胞活性;降低细胞凋亡率、caspase-3的活性及NF-κB蛋白表达,呈浓度依赖性,且对高糖诱导的p65核移位有抑制作用。结论瓜蒌皮提取物对体外高糖诱导的HUVECs凋亡有抑制作用,其作用机制与下调NF-κB表达和降低caspase-3活性有关。Aim To study the inhibitory effects of ex-tractive pericarpium trichosanthes ( EPT) on high glu-cose-induced apoptosis in human umbilical vein endo-thelial cells ( HUVECs ) and its underlining mecha-nisms. Methods HUVECs were cultured. Effects of EPT at different concentrations on the high-glucose-in-duced apoptosis in HUVECs were observed. The cell viability of HUVECs was determined by MTT colori-metric method. Cell apoptosis was identified by Ho-echst staining. The intracellular activity of Caspase-3 was detected with colorimetry. Protein expression and p65 nuclear translocation of NF-κB were detected by Western blot and immunofluorescence staining. Re-sults Treated with 30 mmol · L-1 glucose for 48 hours had a significantly decrease on cell viability com-pared with control, and the apoptotic rate and Caspase-3 activity were increased markedly, the protein expres-sion of NF-κB was upregulated and p65 nuclear trans-location in HUVECs increased; Pre-incubation with EPT(12. 5,25,50 mg·L-1 ) for 1 hour enhanced the cell viability, and decreased the apoptotic rate and Caspase-3 activity and downregulated the expression of NF-κB in a dose-dependent manner. Moreover, EPT could inhibit p65 translocation. Conclusion EPT can protect HUVECs against the apoptosis induced by high glucose in vitro,and its mechanism may be related with downregulation of NF-κB expression and inhibition of the intracellular activity of Caspase-3 .

关 键 词：瓜蒌皮提取物 高糖 内皮细胞 凋亡 NF-κB CASPASE-3 MTT 

分 类 号：R-284.1[医药卫生] R322.123