Effect of nuclear factor-κB p65 ASOND on I typ collagen expression and rat hepatic stellate cells proliferation

( views:, downloads: )
Author:
ZHOU Yu(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
CHEN Ke-quan(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
WANG Hao(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
TANG Zhi-ling(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
WEI Guo li(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
YE Wen-tao(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
YE Shi-cai(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
LIU Rong-huo(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
FENG Xiao(Department of Gastroenterology,Affiliated Hospital of Guangdong Medical College,Zhanjiang Guangdong 524001,China)
Journal Title:
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
Issue:
Volume 14, Issue 06, 2008
DOI:
Key Word:
Oligonucleotides,antisense;Nuclear factor-kappa B;Transfection;Hepatic stellate cells;Ⅰ type collagen;Proliferation;Hepatic fibrosis

Abstract: Objective Sstudy effect of nuclear factor-κB ASOND on I type collagen expression and rat hepatic stellate cells(HSC)proliferation.Methods Rat HSCs were separated by affusing and digestingof Ⅳ type collagenenzyme and density acentric method.Lipid-mediated NF-κB p65 ASOND(0.001,0.01,0.1,1μmol/L)Was transferred into rat HSCs.Toxicity of HSCs caused by NF-κB p65 ASOND and activity of LDH were determined by trypan blue staining.Proliferation affection of transferring NF-κB p65 ASOND into HSCs was determined by MTT.In different concentration NF-κB p65 ASOND.expression of Ⅰ type collagen stimulated by 1mg/L TNF-αwas determined by RT-PCR and ELISA.Results After transfection of NF-κB p65 ASODN,expression of NF-κB protein in HSCs was decreasing.Toxicity experiment indicated that NF-κB p65 ASOND of different concentration(0.001,0.01,0.1 and 1.0 μmol/L)had no effect on HSCs livability and LDH activity(P<0.05).Four different concentration NF-κ B p65 ASOND could restrain HSCs proliferation stimulated by 1 mg/L TNF-α.The expression of I type collagen and mRNA stimulated by 1mg/LTNF-αwas increased,and had a positive correlation with concentration(P>0.05).Conclusion NF-κB p65 ASOND may depress NF-κB activity to restrain HSCs proliferation and Ⅰ type collagen expression,and reduce extracellular matrix.

  • [1]Hellerbrand C,Jobin C,Iimuro Y,et al.Inhibition of NF-κB in activated rat hepatic stellate cells by proteasome inhibitors and an IkappaB super-repressor.Hepatology,1998,27:1285-1295.
  • [2]翁山耕,冷希圣,魏玉华,等.改良法大鼠肝星状细胞的分离培养及鉴定.北京大学学报(医学版),2001,33:83-86.
  • [3]王磊,刘海林,黄欣,等.醛固酮对肝星状细胞Ⅰ、Ⅲ型胶原合成及组织金属蛋白酶抑制因子-1 mRNA表达的作用.中华肝脏病杂志,2004,12:308-310.
  • [4]周宁,唐志凌,魏国丽,等.核因子κB p65反义寡核苷酸对大鼠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.中华肝脏病杂志,2008,16:762-766.
  • [5]Yury P,Eleonora P,Michael B,et al.Halofuginone induces matrix metalloproteinases inrat hepatic stellate cells via activation of p38 and NFkappaB.J Biol Chem,2006,281:15090-15098.
  • [6]Wang Y,Ausman LM,Greenberg AS,et al.Nonalcoholic steatohepatitis induced by a high-fatdiet promotes diethylnitrosamine-initiated early hepatocarcinogenesis in rats.Int J Cancer,2009,124:540-546.
  • [7]Saunder WB.Inhibition of inhibitor of κ B kinases stimulates hepatic stellate cell apoptosis sand accelerated recovery from rat liver fibrosis.Gastroenterology,2005,128:323-326.
  • [8]Planaguma A,Clafia J,Miquel R,et al.The selective cyclooxygenase-2 inhibitor SC-236 reduces liver fibrosis by mechanisms involving non-parenchymal cell apoptosis and PPARgamma activation.FASEB J,2005,19:1120-1122.
WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615 Email:yiyao@wanfangdata.com.cn