全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
-  2015 

bFGF反义寡核苷酸对大鼠肺成纤维细胞增殖信号通路的影响 Role of Antisense Oligonucleotides Basic Fibroblast Growth Factor on the Signaling Pathway of TGF-β_1-Induced Pulmonary Fibroblasts Proliferation

Keywords: bFGF反义寡核苷酸,肺成纤维细胞,增殖,分化,TGF-β1-Smad信号通路

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的:观察碱性成纤维细胞生长因子(bFGF)反义寡核苷酸(AODN)对大鼠肺成纤维细胞增殖、转化信号通路的影响。方法:分离成年大鼠肺成纤维细胞,分为4组:对照组(未转染肺成纤维细胞);TGF-β1组(未转染肺成纤维细胞+5ng/ml TGF-β1刺激);TGF-β1+AODN组(转染AODN肺成纤维细胞+5ng/ml TGF-β1刺激);TGF-β1+RODN(正义寡核苷酸)组(转染RODN肺成纤维细胞+5ng/ml TGF-β1刺激)。每组设3复孔,并作细胞爬片,镜下计数肺成纤维细胞数量,绘制生长曲线。MTT比色法测定肺成纤维细胞增殖率。免疫细胞化学方法检测肌成纤维细胞特异性标志物α-平滑肌肌动蛋白(α-SMA)。通过ELISA法检测培养液中bFGF、CTGF及Ⅰ型胶原蛋白含量。用RT-PCR法分析肺成纤维细胞Smad3 mRNA、Smad7 mRNA、Ⅰ型胶原mRNA表达水平。结果:1细胞计数显示:TGF-β1+RODN组各时段成纤维细胞的数目均较对照组、TGF-β1组、TGF-β1+AODN组明显增多(P<0.05);TGF-β1+AODN组中成纤维细胞增殖数比TGF-β1组明显减低(P<0.05)。2MTT结果显示:TGF-β1+RODN组肺成纤维细胞光密度值明显高于对照组、TGF-β1组、TGF-β1+AODN组(P<0.05);TGF-β1+AODN组中肺成纤维细胞光密度值明显比TGF-β1组减低(P<0.05)。3免疫细胞化学染色结果显示:TGF-β1+RODN组肺成纤维细胞与对照组、TGF-β1组、TGF-β1+AODN组比较,α-SMA染色的平均光密度值(IOD)明显增加(P<0.05);TGF-β1+AODN组中肺成纤维细胞α-SMA染色的平均光密度值(IOD)比TGF-β1组明显减低(P<0.05)。4 ELISA法检测结果显示:在TGF-β1+RODN组肺成纤维细胞培养液中bFGF、CTGF及Ⅰ型胶原蛋白含量明显高于对照组、TGF-β1组、TGF-β1+AODN组(P<0.05);TGF-β1+AODN组肺成纤维细胞培养液中bFGF、CTGF及Ⅰ型胶原蛋白含量比TGF-β1组明显减低(P<0.05)。5RT-PCR法检测结果显示:在TGF-β1+RODN组肺成纤维细胞Smad3mRNA及Ⅰ型胶原蛋白mRNA表达明显高于对照组、TGF-β1组、TGF-β1+AODN组(P<0.05);TGF-β1+AODN组肺成纤维细胞Smad3mRNA及Ⅰ型胶原蛋白mRNA表达明显比TGF-β1组明显减低(P<0.05);TGF-β1+RODN组肺成纤维细胞Smad7mRNA表达明显低于对照组、TGF-β1组、TGF-β1+AODN组(P<0.05);TGF-β1+AODN组肺成纤维细胞Smad7 mRNA表达比TGF-β1组明显增加(P<0.05)。结论:bFGF反义寡核苷酸可抑制肺成纤维细胞的增殖、转化及胶原合成,bFGF反义寡核苷酸可能通过下调TGF-β1-Smad信号通路来发挥作用

References

[1]  Henke C,Fiegel V,Peterson M,et al.Identification and partial characterization of angiogenesis bioactivity in the lower respiratory tract after acute lung injury[J].J Clin Invest,1991,88:1 386-1 395.
[2]  Osaki T,Yoneda K,Tatemoto Y,et al.Peplomycin,a bleomycin derivative,induces myofibroblasts in pulmonary fibrosis[J].Int J Exp Pathol,2001,82(4):231-241.
[3]  Moustakas A,Souehelnytskyi S,Heldin CH.Smad regulation in TGF-beta signal transduction[J].J Cell Sci,2001,114(24):4 359-4 369.
[4]  Lawson WE,Polosukhin VV,Zoia O,et al.Characterization of fibroblast-specific protein 1in pulmonary fibrosis[J].Am J Respir Crit Care Med JT,2005,8(171):899-907.
[5]  Grozio A,Catassi A,Cavalieri Z.Nicotine,lung and cancer[J].Anticancer Agents Med Chem,2007,4(7):461-466.
[6]  Chilosi M,Zamo A,Doglioni C,et al.Migratory marker expression in fibroblast foci of idiopathic pulmonary fibrosis[J].Respiratory Research,2006,7:95.
[7]  Ogata S,Yorioka N,Kohno N.Glucose and prednisolone alter basic fibroblast growth factor expression in peritoneal mesothelial cells and fibroblasts[J].J Am Soc Nephrol,2001,12(12):2 787-2 796.
[8]  Dennler S,Goumans MJ,ten Dijke P.Transforming growth factor-beta signal transduction[J].J Leukoc Biol,2002,71:731-740.
[9]  Verhaeghe C,Tabruyn SP,Oury C,et al.Intrinsic pro-angiogenic status of cystic fibrosis airway epithelial cells[J].Biochem Biophys Res Commun,2007,356(3):745-749.
[10]  Khalil N,Connor R.Idiopathic pulmonary fibrosis:current understanding of the pathogenesis and the status of treatment[J].CMAJ,2004,171:153-160.
[11]  Selman M,Pardo A.Idiopathic pulmonary fibrosis:misunderstandings between epithelial cells and fibroblasts[J]?Sarcoidosis Vasc Diffuse Lung Dis,2004,21:165-172.
[12]  Ward JE,Gould H,Harris T,et al.PPAR gamma ligands,15-deoxy-delta12,14-prostaglandin J2and rosiglitazone regulate human cultured airway smooth muscle proliferation through different mechanisms[J].British Journal of Pharmacology,2004,3(141):517-525.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133