摘要
目的探讨核因子kappa B(NF-κB)p65的小干扰RNA(siRNA)对延迟性排斥反应的抑制作用。方法血管内皮细胞(EOMA)分为7组:A组(空白对照),B组(阴性对照),C组(TNF-α处理),D组(siRNA处理),E组(scramble siRNA处理),F组(TNF-α+siRNA处理),G组(TNF-α+scramble siRNA处理)。用阳离子脂质体LipofectamineTM2000作为转染试剂将化学合成的小鼠NF-κB p65siRNA转染入相应的EOMA细胞,24h后在C组、F组和G组中加入TNF-α(终浓度为20ng/ml)。于TNF-α加入后的6h提取各组细胞总RNA,RT-PCR法测定EOMA细胞内E选择素(E-selectin)、细胞间黏附分子1(ICAM-1)、白细胞介素1α(IL-1α)和血管细胞黏附分子1(VCAM-1)水平。结果 F组EOMA细胞内E-selectin、ICAM-1、IL-1α和VCAM-1表达明显高于A组,的明显低于C组和G组(P<0.01)。结论特异性siRNA抑制NF-κB p65表达可以有效抑制延迟性排斥反应。
Objective To explore the inhibitory effect of nuclear factor kappa B（NF-κB）p65 siRNA on delayed xenograft rejection. Methods The vascular endothelial cells （EOMA） were devided into seven groups of A（blank control, B（negative control）, C（TNF-α treated）, D（siRNA treated）, E（scramble siRNA treated）, F（TNF-α＋siRNA treated）, and G （TNF-α ＋s scramble siRNA treated）. The treatment of TNF-α was performed 24 h after transfection, and 6 h later, the expressions of E- selectin, intercellular adhesion molecule-1 （ICAM-1）, vascular cell adhesion molecule-1 （VCAM-1）, and interleukin-la（IL-lα） mRNA were detected by RT-PCIL Results The mRNA expression levels of E- selectin, ICAM-1 ,VCAM-1, IL-lα were higher in group F than those in group A,but were lower than those in groups of C and G（P〈0. 01）. Conclusion Inhibiting activation of NF-κB p65 by siRNA can efficiently suppress delayed xenograft rejection.
出处
《江苏医药》
CAS
北大核心
2013年第1期1-4,共4页
Jiangsu Medical Journal
基金
国家自然科学基金(39993430)
