核因子κB反义寡核苷酸对转化生长因子β1诱导人肾小管上皮细胞转分化的影响

周磊;王俭勤;宋志霞;曲矿云;孔玉科

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中华肾脏病杂志 ›› 2008, Vol. 24 ›› Issue (7) : 493-497.
基础研究

核因子κB反义寡核苷酸对转化生长因子β1诱导人肾小管上皮细胞转分化的影响

  • 周磊;王俭勤;宋志霞;曲矿云;孔玉科
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Effect of nuclear factor κB antisense oligodeoxynucleotides on transforming growth factor β1-induced epithelial mesenchymal transition in human renal tubular epithelial cells

  • ZHOU Lei, WANG Jian-qin, SONG Zhi-xia, QU Kuang-yun, KONG Yu-ke
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摘要

目的 探讨在转化生长因子β1(TGF-β1)诱导下,核因子κB(NF-κB)反义寡核苷酸对体外培养的人肾小管上皮细胞(HK-2)转分化的影响。 方法 采用脂质体介导的方法将NF-κB反义寡核苷酸(AS-ODN)导入细胞,以TGF-β1(10 μg/L)刺激HK-2细胞24 h后,用RT-PCR方法检测细胞中NF-κB mRNA及α平滑肌肌动蛋白(α-SMA)mRNA表达,用荧光光谱法分析α-SMA蛋白的表达,并以倒置相差显微镜观察细胞转分化过程的形态变化。 结果 TGF-β1诱导24 h后,HK-2细胞中NF-κB mRNA的表达显著上调,为空白对照组的8倍以上(P < 0.01)。NF-κB反义寡核苷酸导入细胞后,可显著抑制TGF-β1诱导的HK-2细胞的 NF-κB mRNA表达,比TGF-β1组减少75%(P < 0.05),同时,α-SMA mRNA和蛋白表达亦较TGF-β1组均明显下调(P < 0.05)。 结论 NF-κB反义寡核苷酸可抑制TGF-β1诱导肾小管上皮细胞NF-κB的表达,抑制肾小管上皮细胞转分化,可能有利于肾间质纤维化的防治。

Abstract

Objective To investigate the effects of nuclear factor κB(NF-κB) antisense oligodeoxynucleotides (AS-ODNs) on transforming growth factor β1(TGF-β1)-induced epithelial mesenchymal transition (EMT) in human renal tubular epithelial cells. Methods NF-κB AS-ODNs were transferred into the human renal tubular epithelial cells (HK-2), and the cells were stimulated by 10 μg/L TGF-β1 for 24 hours. The expression of NF-κB mRNA and α-SMA mRNA were measured by RT-PCR. α-SMA protein expression was assessed by fluorescence spectrum. Results TGF-β1 significantly up-regulated the expression of NF-κB mRNA, which was 8 folds of blank control(P<0.01). TGF-β1-induced epithelial mesenchymal transition was inhibited by NF-κB AS-ODN and the NF-κB mRNA expression of AS-ODNs was decreased by 75%(P<0.05). The expression of α-SMA mRNA and protein was also down-regulated obviously(P<0.05). Conclusion NF-κB AS-ODN can inhibit the expression of NF-κB and the epithelial-mesenchymal transition, which may be a new therapeutic strategy against tubulointerstitial fibrosis.

关键词

核因子κB / 寡核苷酸类 / 反义 / 转化生长因子β / 肾小管 / 上皮细胞 / 转分化

Key words

NF-kappa B / Oligonucleotides / antisense / Transforming growth factor beta / Kidney tubules / Epithelial cells / Transdifferentiation

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周磊;王俭勤;宋志霞;曲矿云;孔玉科. 核因子κB反义寡核苷酸对转化生长因子β1诱导人肾小管上皮细胞转分化的影响[J]. 中华肾脏病杂志, 2008, 24(7): 493-497.
ZHOU Lei;WANG Jian-qin;SONG Zhi-xia;QU Kuang-yun;KONG Yu-ke. Effect of nuclear factor κB antisense oligodeoxynucleotides on transforming growth factor β1-induced epithelial mesenchymal transition in human renal tubular epithelial cells[J]. Chinese Journal of Nephrology, 2008, 24(7): 493-497.
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