米诺环素对人脐静脉内皮细胞-单核细胞黏附的抑制作用及其机制研究

发布时间：2018-02-01 20:10

本文关键词： 米诺环素内皮细胞单核细胞黏附黏附分子转录因子-κB p　出处：《中国药房》2015年31期 　论文类型：期刊论文

【摘要】：目的:研究米诺环素(MC)对肿瘤坏死因子α(TNF-α)诱导内皮细胞-单核细胞黏附的抑制作用及其机制。方法:通过酶消化法从人脐带中获得原代人脐静脉内皮细胞(HUVECs)。将HUVECs细胞分为空白对照组、模型组和MC低、中、高剂量(1、10、100μmol/L)组,药物作用2 h后,除空白对照组外其余各组均加入10 ng/ml TNF-α诱导HUVECs与单核THP-1细胞的黏附,复制内皮细胞-单核细胞黏附模型。荧光显微镜观察黏附细胞数,荧光酶标仪测定其荧光强度;流式细胞仪检测细胞间黏附分子1(ICAM-1)的表达;Western blot法检测细胞核及细胞浆转录因子(NF)-κB p65蛋白的表达。结果:与空白对照组比较,模型组黏附细胞数及其荧光强度、ICAM-1表达、细胞核NF-κB p65蛋白表达均增加,细胞浆NF-κB p65蛋白表达减弱,差异均有统计学意义(P0.01)。与模型组比较,MC低、中、高剂量组黏附细胞数及其荧光强度、ICAM-1表达均降低;MC中、高剂量组细胞核NF-κB p65蛋白表达减弱,细胞浆NF-κB p65蛋白表达增强,差异均有统计学意义(P0.01或P0.05)。结论:MC能显著地减少TNF-α诱导的内皮细胞-单核细胞的黏附,其机制可能与降低HUVECs细胞表面ICAM-1的表达、抑制NF-κB p65蛋白的表达有关。
[Abstract]:Objective: to study the effect of minocycline (MMC) on tumor necrosis factor 伪 (TNF- 伪). Inhibitory effect of Endothelial Cell-Monocyte Adhesion and its Mechanism. Methods: primary Human umbilical vein Endothelial cells (HUVECs) were obtained from Human umbilical Cord by Enzymatic digestion. HUVECs cells were divided into blank control group. The model group and the low, medium and high dose of MC group (100 渭 mol / L) were treated with the drug for 2 h. 10 ng/ml TNF- 伪 was used to induce the adhesion of HUVECs to mononuclear THP-1 cells except the blank control group. Endothelial cell-monocyte adhesion model was established. The number of adherent cells was observed by fluorescence microscope and the fluorescence intensity was measured by fluorescence enzyme labeling instrument. The expression of ICAM-1 was detected by flow cytometry. Western blot method was used to detect the expression of NF 魏 B p65 protein in nucleus and cytoplasm. Results: compared with the control group, the number of adhesion cells and the fluorescence intensity of the model group were measured. ICAM-1 expression, nuclear NF- 魏 B p65 protein expression increased, and cytoplasm NF- 魏 B p65 protein expression decreased. Compared with the model group, the number of adhesion cells and the expression of ICAM-1 in the high dose group were lower than those in the model group. In MC, the expression of NF- 魏 B p65 protein was decreased and the expression of NF- 魏 B p65 protein in cytoplasm was increased in high dose group. Conclusion the TNF- 伪 -induced endothelial cell-monocyte adhesion can be significantly reduced by the addition of TNF- 伪. The mechanism may be related to the reduction of ICAM-1 expression on HUVECs cells and the inhibition of NF- 魏 B p65 protein expression.
【作者单位】：苏州市食品药品检验所;
【分类号】：R96
【正文快照】： *主管药师。研究方向:药学。电话:0512-68222721。E-mail:chenli7710@126.com血管内皮细胞的损伤在动脉粥样硬化(AS)形成过程中具有重要作用[1]。在AS形成早期,损伤的内皮细胞会促进单核细胞募集和黏附,单核细胞随后穿过内膜迁移至内膜下分化成巨噬细胞,摄取氧化低密度脂蛋白

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