摘要
目的 ：细胞间黏附分子（intercellular adhesion molecule,ICAM）-1在炎症性肠病患者肠道炎症的发生中起着十分重要的作用。肿瘤坏死因子（tumor necrosis factor,TNF）-α可诱导内皮细胞ICAM-1的过度表达。本研究通过姜黄素干预人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVEC）,观察姜黄素是否能够影响TNF-α诱导ICAM-1的表达。方法：从新生儿脐带中获取HUVEC进行原代培养,取第3～5代的细胞,并将其分成3个实验组：空白对照组,不做处理;TNF-α组,加入10 ng/ml TNF-α干预3 h;姜黄素组,先加入10μmol/L姜黄素干预2 h,再加入10 ng/ml TNF-α干预3 h。通过流式细胞仪检测3组HUVEC细胞表面ICAM-1的表达量;同时通过免疫荧光技术观察3组细胞表达ICAM-1的荧光强度;Real-time PCR技术检测3组细胞中ICAM-1 mRNA的表达。结果：①流式细胞检测发现,TNF-α组中HUVEC表面ICAM-1的表达较空白对照组明显增加[（88.69±3.14）%vs（9.82±1.21）%,P〈0.01];姜黄素组中ICAM-1表达[（41.85±8.39）%]较TNF-α组明显下降（P〈0.01）,但高于空白对照组（P〈0.01）;②免疫荧光检测也显示,TNF-α组中HUVEC细胞表面ICAM-1的表达强度明显高于空白对照组,而姜黄素组ICAM-1的表达强度较TNF-α组减弱。③Real-time PCR显示,TNF-α组中ICAM-1 mRNA的表达较空白对照组增加（34.70±14.99 vs 1.03±0.26,P〈0.05）;姜黄素组中ICAM-1 mRNA表达（15.34±8.42）较TNF-α组下降（P〈0.01）,比空白对照组增加（P〈0.05）。结论：姜黄素可抑制TNF-α诱导的HUVEC表面ICAM-1蛋白的表达,这与其能够抑制细胞内ICAM-1 mRNA的表达有关。
Objective：Intercellular adhesion molecule（ICAM1）-1 plays a very important role in intestinal inflammation of patients with inflammatory bowel disease. Tumor necrosis factor-α（TNF-α） induces overexpression of ICAM-1 in endothelial cells. The aims of study were to investigate the effects of curcumin on human umbilical vein endothelial cells（HUVEC） and to observe whether curcumin can influence expression of ICAM-1 induced by TNF-α. Methods：HUVECs were obtained from umbilical cord of newborns and cultured and generated from primary to the fifth generation. The third to fifth generation of cells were selected and divided into three experimental groups,including control group,TNF-α group with 10 ng / ml TNF-α intervention for 3 hours and curcumin group.In curcumin group,HUVEC was cultured with 10 μmol / L curcumin for 2 hours and then continued to culture for 3 hours after adding10 ng / ml TNF-α. The expression of ICAM-1 was detected by flow cytometry and immunofluorescence. Flow cytometry was performed to detect the expression of ICAM-1 on HUVEC surface in the three groups. Meanwhile,immunofluorescence was performed to observe the fluorescence intensity of the expression of ICAM-1. The expression of ICAM-1 mRNA was measured by Real-time PCR. Results：①The expression of ICAM-1 in HUVEC in the TNF-α group was significantly increased than that in the control group [（88.69 ±3.14）% vs（9.82 ± 1.21）%,P〈0.01]. The expression of ICAM-1 in the curcumin group [（41.85 ± 8.39）%] was significantly decreased than that in the TNF-α group（P〈0.01）,but higher than that in the control group（P〈0.01）. ②The immunofluorescence results also showed that the expression intensity of ICAM-1 in the TNF-α group was significantly higher than that in the control group.The expression intensity of ICAM-1 in the curcumin group was decreased than that in the TNF-α group. ③The expression of ICAM-1mRNA in the TNF-α group was increased than that in the control group（34.70 ± 14.99 vs 1.03 ± 0.26,P〈0.05）. The expression of ICAM-1 mRNA in the curcumin group（15.34 ± 8.42） was significantly decreased than that in the TNF-α group（P〈0.01）,but higher than that in the control group（P〈0.05）. Conclusion：The results showed that curcumin could inhibit the expression of ICAM-1protein in HUVEC induced by TNF-α. The effect may be related to the inhibition of ICAM-1 mRNA expression.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第11期1476-1480,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
南京市医学科技发展项目(ZKX10014)
关键词
细胞间黏附分子-1
肿瘤坏死因子-α
人脐静脉内皮细胞
炎症性肠病
intercellular adhesion molecule-1
tumor necrosis factor-α
human umbilical vein endothelial cells
inflammatory bowel disease
