核因子-κBp65反义寡核苷酸对肝纤维化作用的实验研究  

The experimental study on effects of nuclear factor-κBp65 antisense oligonucleotide on liver fibrosis

作　　者：周宇[1] 陈科全[1] 叶石才[1] 王壮[1] 梁坚[1] 刘荣火[1] 唐志凌[1] 王皓[1] 冯晓[1] 叶文桃[1] 

机构地区：[1]广东医学院附属医院消化科,湛江524001

出　　处：《中华消化杂志》2009年第4期254-257,共4页Chinese Journal of Digestion

基　　金：广东省自然基金资助项目（04011392）

摘　　要：目的探讨核因子（NF）-κBp65反义寡核苷酸（ASODN）对肝星状细胞（HSC）NF-κB活性和白细胞介素（IL）-6表达的影响。方法分离培养大鼠HSC，用锥虫蓝染色法和电泳迁移位移试验（EMSA）分别检测NF-κBp65 ASODN对HSC毒性和NF-κB活性的影响，RT—PCR法和ELISA法分别检测对IL-6mRNA和蛋白表达的影响。结果ASODN在0．001～1．0μmol／L浓度时，对体外培养的HSC无明显毒性。HSC经肿瘤坏死因子（TNF）-α刺激后，NF-κB活性增强，在0．001～1．0μmol／L的ASODN作用后，NF-κB活性明显减弱，且呈剂量依赖性。同时转染ASODN（0．001～1．0μmol／L）后，TNF-α诱导HSC的IL-6 mRNA及蛋白表达明显降低，亦呈剂量依赖性。结论ASODN能特异性降低NF-κB活性，同时减少HSC的IL-6表达，为其治疗肝纤维化提供了理论基础。Objective To investigate the effect of the nuclear factor （NF）-κBp65 antisense oligonucleotide （ASODN） on NF-κB activity and expression of interleukin（IL）-6 in hepatic stellate cells （HSC）. Methods The HSC were separated from rats and cultured. The toxicity of NF-κBp65 ASODN on HSC were detected by Trypan blue exclusion staining and the NF-κB activity was determined by EMSA. The expressions of IL-6 mRNA and protein were meaured by RT-PCR and ELISA, respectively. Results In vitro, no toxicity of ASODN on HSC was observed at the concentrations of 0. 001 to 1. 0 μmol/L. NF-κB activity was increased after stimulating HSC with tumor necrosis factor （TNF）α, whereas it was weakened in a dose dependent manner when HSC were cultured with ASODN （concentration from 0. 001 to 1.0μmol/L）. At the same time, the expressions of IL-6 mRNA and protein induced by TNFa were decreased after transfected with ASODN at concentrations of 0. 001- 1. 0μmol/L in a dose dependent manner. Conclusion ASODN may specifically inhibit either the activiy of NF-κB or expression of IL-6, which provides the theoretical basis that ASODN may use to treat fibrosis of the liver.

关 键 词：肝硬化 核因子-ΚB 肝星状细胞 白细胞介素6 

分 类 号：R686[医药卫生—骨科学]