摘要
目的探讨二苯乙烯苷(TSG)对肿瘤坏死因子α(TNF-α)诱导的内皮细胞损伤的保护性作用及其对内皮细胞NF-κB核转位的影响。方法取新生儿脐带分离原代人脐静脉内皮细胞(HUVEC),用因子Ⅷ鉴定内皮细胞。按照分组,HUVEC分别与TNF-α60μg·L-1,TNF-α+TSG 1,10,25,50和100μmo·lL-1进行孵育,TSG预孵24 h,再加TNF-α处理3 h。显微镜观察TSG对TNF-α诱导HUVEC的保护性作用,MTT法检测细胞存活率,免疫荧光法检测细胞NF-κB的核转位现象。结果因子Ⅷ相关抗原检测显示,分离的细胞为内皮细胞。显微镜观察显示,TSG对TNF-α诱导的HUVEC损伤具有明显的抑制作用,TSG 100μmo·lL-1使内皮细胞形态几乎接近正常组水平。MTT结果显示,TSG可显著提高TNF-α诱导后的细胞存活率,并呈浓度依赖性(r=0.9657,P<0.01)。免疫荧光检测表明,TNF-α诱导NF-κB发生核内转移,TSG作用后可抑制这种核转位,TSG 100μmol·L-1处理组的核转位抑制作用最明显。结论TSG可能通过抑制NF-κB的激活发挥保护血管内皮的作用。
OBJECTIVE To investigate the protective effect of 2,3,5,4＇-tetrahydroxystilbene-2-O-β-Dglucoside（ TSG） on human umbilical vein endothelial cells（ HUVECs） and on NF-κB nuclear translocation. METHODS HUVECs were isolated from the newborn umbilical cord and identified by factor Ⅷ. They were incubated with TNF-α 60 μg·L^- 1,and TNF-α ＋ TSG 1,10,25,50 and 100 μmol·L^- 1,respectively. They were treated with TSG for 24 h before treatment with TNF-α for anthor 3 h. Protective effects of TSG on HUVECs induced by TNF-α were observed under a microscope. Cell viability of HUVECs was analyzed by MTT assay. NF-κB nuclear translocation was tested by immunofluorescence.RESULTS Factor Ⅷ analysis revealed that the isolated cells were HUVECs. Microscopic observation indicated that TSG significantly inhibited TNF-α-induced dysfunction in HUVECs. The cell morphology of the group pretreated with TSG 100 μmo·lL^- 1was similar to that of the normal group. MTT analysis indicated that TSG notably improved cell viability after treatement with TNF-α in a concentration-dependent manner（ r = 0. 9657,P 0. 01）. The immunofluorescence results showed that TSG inhibited NF-κB nuclear translocation induced by TNF-α. TSG 100 μmol·L^- 1had the most obvious inhibitory effect on NF-κB nuclear translocation induced by TNF-α. CONCLUSION TSG can protect vascular endothelial cells by inhibiting NF-κB activation.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2014年第1期24-28,共5页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金资助项目(81202879)
南通大学人才引进启动费项目(122091)
南通市应用研究计划(BK2012087)
南通市科技计划项目(BK2011044)~~
关键词
二苯乙烯苷
动脉粥样硬化
NF-ΚB
肿瘤坏死因子Α
人脐静脉内皮细胞
核转位
2
3
5
4＇-tetrahydroxystilbene-2-O-β-D-glucoside
atherosclerosis
NF-κB
tumor necrosis factor-α
human umbilical vein endothelial cells
nuclear translocation
