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《Journal of Sun Yat-sen University (Medical Sciences)》 2005-01
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Expression of Fractalkine in Renal Tubular Epithelial Cells Mediate Chemotaxis of Macrophages

TANG Ke-jing, LI You-ji, XIE Can-mao, HUANG Shun-wei, LI Xiao-yan, YU Xue-qing(Department of Nephrology, The First Affiliated Hospital, SUN Yat-sen University, Guangzhou 510080, China)  
To investigate if tumor necrosis factor-alpha (TNF-α) could induce NRK-52E cells, a rat renal proximal tubular epithelial cell line, up-regulating expression of fractalkine, one of chemokines, and if the induced effect appear time- and dose-dependent, and at the same time to investigate if the expression of fractalkine in renal tubular epithelial cells mediate macrophages chemotaxis. Using immunofluorescence cytochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis to detect the effect of TNF-α on the expression of fractalkine in NRK-52E cells, and using Transwell to investigate the chemotaxis of RAW 264.7 cells, a murine monocyte/macrophage cell line, to NRK-52E cells, and to investigate the effect of anti-fractalkine neutralizing antibody to the chemotaxis. Normal cultured NRK-52E cells could express few fractalkine. Incubation of NRK-52E cells with 40 ng/ml TNF-α for 1 hour did not up-regulate fractalkine mRNA and protein expression significantly. The evident effect of TNF-α on NRK-52E cells fractalkine mRNA and protein expression was observed at 6 h and the maximal effect at 24 h to 48 h. We observed that 10 ng/mL TNF-α had the effect to up-regulate fractalkine mRNA and protein expression, and the concentrations of20 ng/mL or 40 ng/mL TNF-α had more stronger effect. The up-regulated expression of fractalkine in NRK-52E cells could enhance the chemotaxis of macrophages significantly, but this chemotaxis could be inhibited dramatically using anti-fractalkine neutralizing antibody. [Conclusion] The mRNA and protein expression of fractalkine in NRK-52E cells were up-regulated by TNF-α in a time- and dose-dependent manner, and fractalkine played an important role in mediating the chemotaxis of macrophages to renal tubular epithelial cells.
【Fund】: 教育部博士点科研基金资助项目(2000056916)
【CateGory Index】: R692
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【References】
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【Secondary References】
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