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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 m RNA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 m RNA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 m RNA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 m RNA的表达有关。
Objective: Intercellular adhesion molecule (ICAM)-1 plays an important role in the development of intestinal inflammation in patients with inflammatory bowel disease. Tumor necrosis factor (TNF)-α can induce overexpression of ICAM-1 in endothelial cells. In this study, curcumin was administered to human umbilical vein endothelial cells (HUVEC) to observe whether curcumin could affect the expression of ICAM-1 induced by TNF-α. METHODS: HUVECs were obtained from the umbilical cord of neonates for primary culture. Cells from the 3rd to 5th passages were collected and divided into 3 experimental groups: blank control group and no treatment. TNF-α group was added with 10 ng/ml. TNF-α was intervened for 3 h; curcumin group was treated with 10 μmol/L curcumin for 2 h, and 10 ng/ml TNF-α was added for 3 h. The expression of ICAM-1 on the surface of three groups of HUVEC cells was detected by flow cytometry. The fluorescence intensity of ICAM-1 was also observed in three groups of cells by immunofluorescence; the ICAM-1 mRNA in three groups of cells was detected by Real-time PCR. expression. RESULTS:1 Flow cytometry showed that the expression of ICAM-1 on HUVEC surface in TNF-α group was significantly higher than that in blank control group [(88.69±3.14)% vs (9.82±1.21)%, P<0.01]; Curcumin group The expression of ICAM-1 in the medium [(41.85±8.39)%] was significantly lower than that in the TNF-α group (P<0.01), but higher than that in the blank control group (P<0.01). 2 Immunofluorescence assay also showed that in the TNF-α group The intensity of ICAM-1 expression on HUVEC cells was significantly higher than that of the blank control group, while the expression intensity of ICAM-1 in the curcumin group was weaker than that in the TNF-α group. 3Real-time PCR showed that the expression of ICAM-1 mRNA in the TNF-α group was higher than that in the blank control group (34.70±14.99 vs 1.03±0.26, P<0.05); the expression of ICAM-1 mRNA in the curcumin group was 15.34± 8.42) decreased compared with TNF-α group (P<0.01) and increased compared with blank control group (P<0.05). CONCLUSION: Curcumin can inhibit the expression of ICAM-1 protein on HUVEC surface induced by TNF-α, which is related to its ability to inhibit the expression of ICAM-1 mRNA in cells.