核因子-KB p65反义寡核苷酸对大鼠肝星状细胞转化生长因子β1及细胞间黏附分子1表达的影响  

Effects of nuclear factor-kappa B p65 ASODN on transforming growth beta-1 and intercellular adhesion molecule-1 of rat hepatic stellate ceils

作　　者：周宇[1] 唐志凌[1] 魏国丽[1] 王皓[1] 叶石才[1] 陈科全[1] 陈元鸿[1] 刘荣火[1] 

机构地区：[1]广东医学院附属医院消化内科,湛江524001

出　　处：《中华肝脏病杂志》2008年第10期762-766,共5页Chinese Journal of Hepatology

基　　金：广东省自然科学基金（04011392）

摘　　要：目的探讨核因子-KB p65反义寡核苷酸（NF-KB p65 ASODN）对大鼠肝星状细胞HSC炎症因子转化生长因子β1（TGFβ1）及细胞间黏附分子1（ICAM—1）表达的影响。方法Ⅳ型胶原酶消化密度梯度离心法分离培养大鼠HSC；人工合成NF-KB p65 ASODN并行全程硫代磷酸化修饰；观察脂质体介导的不同浓度的NF—KB p65 ASODN（0．001、0．010、0．100、1．000μmol／L）对肿瘤坏死因子（TNF）q刺激HSC产生TGFβ1及ICAM-1 mRNA和蛋白质表达的影响，蛋白凝胶电脉迁移率变动分析法检测NF-KB p65 ASODN对TNFβ1刺激HSC NF-kB的活性影响。结果TNFβ1刺激HSC后，NF—KB活性和TGFβ1、ICAM-1的表达明显增强；应用NF—KB p65 ASODN（0．001～1．000μmol／L）作用后，NF—KB活性明显减弱，且呈剂量依赣洼，灰度值分别为16070±223，15715±199，14999±224，14447±228，各组比较，p值均〈0．05。同时TGFβ1、ICAM1的mRNA及蛋白质表达明显减少，亦呈剂量依赖性，P值均〈0．05。结论NF—KB p65 ASODN可特异性降低HSC的NFKB的活性，明显抑制TGFβ1、ICAM-1的表达，为NF-KB p65 ASODN治疗肝纤维化提供了理论依据。Objective Objectives To study the effects of nuclear factor （NF）-kappa B p65 ASODN on transforming growth factor beta-1 （TGF beta 1） and intercellular adhesion molecule-1 （ICAM-1） of rat hepatic stellate cells （HSC） and the mechanisms of NF-kappa B p65 ASODN in treating liver fibrosis. Meth- ods Type IV collagen enzyme digestion and density centrifugation methods were used to separate rat hepatic stellate cells. NF-kappa B p65 ASODN was manually synthesized and completely phosphorothioate-modified. The changes of TGF beta 1 and ICAM-1 mRNA were detected by RT-PCR and albumen of TGF beta 1 and ICAM-1 were detected by ELISA. The changes ofNF-kappa B activity were determined by ELISA. Results NF-kappa B activity and the expressions of ICAM-1 and TGF beta 1 increased after the HSC were trcated by TNF alpha, NF-kappa B activity weakened after being treated with NF-kappa B p65 ASODN （0.001-1.000μmol/L）, P 〈 0.05 in a dose dependent manner. Transferring NF-kappa B p65 ASODN （0.001-1.000 pmol/L） also weakened the expression of ICAM-1 and TGF beta 1 mRNA and the protein induced by TNF alpha in HSC. It was also in a dose dependent manner, P 〈 0.05. Conclusions After transferring NF-kappa B p65 ASODN into HSC, their NF-kappa B activity decreased, and their mRNA and protein expressions of ICAM- 1 and TGFbeta 1 also decreased. This may serve as a new way in treating hepatic fibrosis.

关 键 词：肝纤维化 核因子KB 反义寡核苷酸 肝星状细胞 转化生长因子Β1 细胞间黏附分子1 

分 类 号：R575.2[医药卫生—消化系统]