E-选择素和细胞间粘附分子-1反义寡核苷酸对人脐静脉内皮细胞与单核细胞粘附功能的影响

发布时间：2018-04-25 14:30

  本文选题：E-选择素 + 细胞间粘附分子-1　； 参考：《福建医科大学》2008年硕士论文

【摘要】： 目的:探讨E-选择素(E-selectin)和细胞间粘附分子-1(ICAM-1)反义寡核苷酸(asODN)及其与辛伐他汀(simvastatin)联合应用对人脐静脉内皮细胞(HUVEC) ICAM-1和E-selectin表达的抑制和对外周血单个核细胞粘附的影响。 方法:原代培养人脐静脉内皮细胞(HUVEC),并传代;应用脂质体转染法将人工合成E-selectin asODN和ICAM-1 asODN分别导入HUVECs;实验共分8组,分别为空白对照(basal)组、TNF-α诱导组、liposome对照组、liposome-control asODN组、裸asODN (nude asODN)组、脂质体asODN (lipo-asODN)组、simvastatin处理组、lipo-asODN与simvastatin联合组,除空白组外,其余7组均以TNF-α诱导内皮损伤;以流式细胞技术测定HUVECs细胞膜E-selectin和ICAM-1蛋白的表达;以RT-PCR半定量技术测定HUVECs E-selectin mRNA和ICAM-1 mRNA的表达;以细胞粘附试验测定HUVECs与单个核细胞的粘附率;应用辛伐他汀预先处理HUVECs,再用E-selectin lipo-asODN和ICAM-1 lipo-asODN转染HUVECs,观察E-selectin asODN和ICAM-1 asODN与辛伐他汀联合应用对HUVECs E-selectin和ICAM-1的表达以及对HUVECs与外周血单个核细胞粘附抑制的影响。 结果:⑴与TNF-α诱导组相比,E-selectin asODN和ICAM-1 asODN以及E-selectin lipo-asODN和ICAM-1 lipo-asODN均明显降低了HUVECs相应的E-selectin和ICAM-1蛋白及mRNA的表达(P0.01);⑵E-selectin asODN和ICAM-1 lipo-asODN与simvastatin联合应用可降低对HUVECs相应的E-selectin和ICAM-1的蛋白及mRNA表达;⑶lipo-asODN与simvastatin联合组与1ipo-asODN组比较,E-selectin表达水平显著下降(P0.01),而ICAM-1的表达水平下降无显著意义(P0.05),表明E-selectin lipo-asODN与simvastatin联合作用有更好的抑制效果。⑷与TNF-α刺激组相比,lipo-asODN组及1ipo-asODN与simvastatin联合组HUVECs与单个核细胞粘附率均显著下降(P0.01),尤其以HUVECs与CD3+T淋巴细胞的粘附率下降最为明显,但1ipo-asODN与simvastatin联合组与1ipo-asODN组比较,未见明显变化(P0.05)。 结论:E-selectin asODN和ICAM-1 asODN可抑制HUVECs相应的蛋白及mRNA的表达,并可抑制HUVECs与单个核细胞的粘附,尤其对HUVECs与CD3+ T细胞粘附的抑制作用更为明显。1ipo-asODN与simvastatin联合应用比单独1ipo-asODN的抑制作用更加明显。本研究为E-selectin asODN和ICAM-1 asODN应用于临床抗动脉粥样硬化的基因治疗的前景开拓新的思路和提供理论依据。
[Abstract]:Aim: to investigate the inhibition of ICAM-1 and E-selectin expression in human umbilical vein endothelial cells (HUVECs) by Eselectin E-selectin (E-selectin) and intercellular adhesion molecule-1 (ICAM-1) antisense oligodeoxynucleotides (ASODN) and its combination with simvastatin in human umbilical vein endothelial cells (HUVECs) and the effects on the adhesion of peripheral blood mononuclear cells (PBMC). Methods: human umbilical vein endothelial cells (HUVECs) were cultured and subcultured, the synthetic E-selectin asODN and ICAM-1 asODN were transfected into HUVECs by liposome transfection, and were divided into 8 groups: TNF- 伪 induced liposome-control asODN group and naked asODN nude asODN group. The endothelial damage was induced by TNF- 伪 in the liposome asODN liposome lipo-asODN group and simvastatin treated group, and the expression of E-selectin and ICAM-1 protein in the HUVECs cell membrane was measured by flow cytometry. The expression of HUVECs E-selectin mRNA and ICAM-1 mRNA was detected by RT-PCR semi-quantitative technique, and the adhesion rate of HUVECs to mononuclear cells was measured by cell adhesion test. HUVECs were pretreated with simvastatin and transfected with E-selectin lipo-asODN and ICAM-1 lipo-asODN. The effects of E-selectin asODN and ICAM-1 asODN combined with simvastatin on the expression of HUVECs E-selectin and ICAM-1 and the inhibition of HUVECs adhesion to peripheral blood mononuclear cells were observed. Results compared with TNF- 伪 induced group, the expression of E-selectin asODN and ICAM-1 asODN, E-selectin lipo-asODN and ICAM-1 lipo-asODN, E-selectin and ICAM-1 protein of HUVECs and the expression of mRNA, P0.01E-selectin asODN and ICAM-1 lipo-asODN combined with simvastatin decreased the corresponding E-selectin and ICAM-1 protein and mRNA expression of HUVECs. The expression of E-selectin in 3lipo-asODN combined with simvastatin group was significantly lower than that in 1ipo-asODN group, while the expression level of ICAM-1 was not significantly decreased in 1ipo-asODN group. It indicated that the combined effect of E-selectin lipo-asODN and simvastatin was better than that of TNF- 伪 stimulation group. 4. The inhibitory effect was better than that of TNF- 伪 -stimulated group and 1ipo-asODN and simvastatin group. In the combined group, the adhesion rate of HUVECs to mononuclear cells decreased significantly, especially the adhesion rate of HUVECs to CD3 T lymphocytes. However, there was no significant difference between 1ipo-asODN and simvastatin group and 1ipo-asODN group. Conclusion asODN and ICAM-1 asODN can inhibit the expression of HUVECs protein and mRNA, and inhibit the adhesion of HUVECs to mononuclear cells. In particular, the inhibitory effect on HUVECs adhesion to CD3 T cells was more obvious. 1Ipo-asODN combined with simvastatin had more obvious inhibitory effect than 1ipo-asODN alone. This study provides new ideas and theoretical basis for the application of E-selectin asODN and ICAM-1 asODN in gene therapy against atherosclerosis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;R543

【参考文献】

相关期刊论文 前9条

1 彭创;汤恢焕;;反义寡核苷酸在恶性肿瘤治疗中的作用及机制[J];湘南学院学报(医学版);2007年03期

2 周娟;崔玉霞;杨锡强;王莉佳;蒋利萍;;特异性反义寡核苷酸抗小鼠呼吸道合胞病毒感染的研究[J];第三军医大学学报;2007年02期

3 张向阳;卢启明;;Survivin反义寡核苷酸诱导胃癌SGC7901细胞凋亡及对白藜芦醇的增敏作用[J];第四军医大学学报;2007年01期

4 唐可京;ICAM-1和VCAM-1的结构与表达调控[J];国外医学(分子生物学分册);2002年03期

5 牛俊奇;汪杨;;病毒性肝炎基因治疗研究的现状[J];内科理论与实践;2007年04期

6 李东良;吕联煌;林振兴;陈英玉;;新型bcl-2基因反义寡核苷酸F951诱导人白血病HL60细胞凋亡[J];中国药理学通报;2007年02期

7 张敏;袁方;陈晖;方唯一;;辛伐他汀对CD40L介导的ECV-304细胞E选择素表达及粘附功能的影响[J];中国药理学通报;2007年12期

8 ;Inhibitory effect of antisense oligodeoxynucleotide to p44/p42 MAPK on angiotensin Ⅱ-induced hypertrophic response in cultured neonatal rat cardiac myocyte[J];Acta Pharmacologica Sinica;2004年01期

9 李远方,胡健;霉酚酸酯减少黏附分子P-选择素和ICAM-1在鼠主动脉表达并抑制粥样硬化病变形成[J];中国医科大学学报;2005年02期

，

本文编号：1801715