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免疫荧光检测核因子-κB活化的定量分析 被引量:1

Quantitative Analysis of Immuno-fluorescence of Nuclear Factor-κB Activation
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摘要 免疫荧光可定性分析细胞内蛋白核转位现象,NF-κB/p65蛋白核转位提示NF-κB信号通路的活化,我们尝试运用自主研发设计的免疫荧光分析软件计算荧光图像中胞核和胞质位置的荧光值变化,定量分析核转位情况,并通过蛋白质免疫印迹法验证软件分析结果。应用该软件分析内毒素诱导0.5、1、2和4h后原代人脐静脉内皮细胞的NF-κB信号通路活化,结果提示2h为核转位高峰期,与蛋白质免疫印迹法验证结果一致,表明自主研发的免疫荧光分析软件可应用于免疫荧光的定量分析。 Immuno-fluorescence technique can qualitatively determine certain nuclear translocation,of which NF-κB/p65 implicates the activation of NF-κB signal pathways.Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm.We verified the quantitative analysis by Western Blot.When we applied the software to analysis of nuclear translocation in lipopolysaccharide(LPS)induced(0.5h,1h,2h,4h)primary human umbilical vein endothelial cells(HUVECs),we found that nuclear translocation peak showed up at 2has with calculated Western blot verification results,indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
出处 《生物医学工程学杂志》 EI CAS CSCD 北大核心 2015年第3期669-674,共6页 Journal of Biomedical Engineering
基金 国家自然科学基金资助项目(81160193) 江西省卫生厅科技计划资助项目(20121053)
关键词 免疫荧光 荧光定量分析 NF-ΚB 人脐静脉内皮细胞 内毒素 immuno-fluorescence fluorescence quantitative analysis NF-kappa B human umbilical vein endothelialcells lipopolysaeeharide
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参考文献16

  • 1TIERI P, TERMANINI A, BELLAVISTA E, et al. Char- ting the NF-kB pathway interactome map [J]. PLoS One, 2012, 7(3): e32678.
  • 2ROGERS J A, FUSELER J W. Regulation of NF-kappaB ac- tivation and nuclear translocation by exogenous nitric oxide (NO)donors in TNF-alpha activated vascular endothelial ceils [J]. Nitric Oxide, 2007, 16(3): 379-391.
  • 3GUZMAN J R, KOO J S, GOLDSMITH J R, et al. Oxymatrine prevents NF-kB nuclear translocation and ameliorates acute intes- tinal inflammation[J]. Sei Rep, 2013, 3: 1629.
  • 4KADAM S S, TIWARI S, BHONDE R R. Simultaneous iso- lation of vascular endothelial ceils and mesenehymal stem cells from the human umbilical cord [J]. In Vitro Cell Dev Biol Anita, 2009, 45(1/2): 23-27.
  • 5SIOW R C. Culture of human endothelial cells from umbilical veins[J]. Methods Mol Biol, 2012, 806 : 265-274.
  • 6WANG P, CHEN R, LUO W, et al. Research and imple- mentation of conglutinated macrophage image segmentation based on improved watershed algorithm [J].Adv Mat Res, 2011, 225: 483-487.
  • 7王平,张力,周长其.基于种子点的粘连巨噬细胞图像的分割方法[J].液晶与显示,2012,27(6):808-813. 被引量:5
  • 8游迎荣,范影乐,庞全.基于距离变换的粘连细胞分割方法[J].计算机工程与应用,2005,41(20):206-208. 被引量:32
  • 9SWEENEY C, LI L, SHANMUGAM R, et al. Nuclear fac- tor-κB is constitutively activated in prostate Cancer in vitro and is overexpressed in prostatic intraepithelial neoplasia and adenocarcinoma of the prostate [J]. Clin Cancer Res, 2004, 10(16) : 5501-5507.
  • 10HELLMAN L M, FRIED M G. Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid interac- tions [J]. Nat Protoc, 2007, 2(8) : 1849-1861.

二级参考文献53

  • 1林巧爱,陈向敏,夏克栋,董海燕,陈韶,张丽芳.NF-κB、COX-2在宫颈癌组织中的表达及其与HPV16感染的关系[J].癌变.畸变.突变,2007,19(2):113-115. 被引量:10
  • 2Ingber D. How cells (might) sense microgravity [ J]. FASEB J, 1999, 13(Suppl) :3 - 15.
  • 3Qian AR, Hu LF, Gao X, et al. Large gradient high magnetic field affects the association of MACF1 with actin and microtubnle cytoskeleton [ J]. Bioelectromagnetics, 2009, 30 ( 7 ) : 545 -555.
  • 4Carmeliet G, Nys G, Bouillon R. Microgravity reduces the differentiation of human osteoblastic MG-63 cells [ J ]. J Bone Miner Res, 1997, 12(5) :786 -794.
  • 5Doty SB, Morey-Holton ER, Dumova GN, et al. Cosmos 1887: morphology, histochemistry, and vasculature of the growing rat tibia [J]. FASEB J, 1990, 4(1):16 -23.
  • 6Hughes-Fulford M, Rodenacker K, Jutting U. Reduction of anabolic signals and alteration of osteoblast nuclear morphology in microgravity [J]. J Cell Biochem, 2006, 99(2):435-449.
  • 7Braun M, Sievers A. Role of the microtubule cytoskeleton in gravisensing Chara rhizoids [J]. Eur J Cell Biol, 1994, 63(2) :289 -298.
  • 8Montague PR, Friedlander MJ. Morphogenesis and territorial coverage by isolated mammalian retinal ganglion cells [ J ]. J Neurosci, 1991, 11(5) :1440 -1457.
  • 9Gazit Y, Baish JW, Safabakhsh N, et al. Fractal characteristics of tumor vascular architecture during tumor growth and regression [ J]. Microcirculation, 1997, 4 (4) : 395 - 402.
  • 10Caldwell CB, Moran EL, Bogoch ER. Fractal dimension as a measure of altered trabecular bone in experimental inflammatory arthritis [J]. J Bone Miner Res, 1998, 13(6) :978 -985.

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