CAJ | 학술논문

目的探讨在转化生长因子β1(TGF-β1)诱导下,核因子kB(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-kB反义寡核苷酸(AS-ODN)导入细胞,以TGF-μ1(10 μg/L)刺激HK-2细胞24 h后,用RT-PCR方法检测细胞中NF-kB 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-kB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-kB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.
목적 탐토재전화생장인자β1(TGF-β1)유도하,핵인자kB(NF-κB)반의과핵감산대체외배양적인신소관상피세포(HK-2)전분화적영향.방법 채용지질체개도적방법장NF-kB반의과핵감산(AS-ODN)도입세포,이TGF-μ1(10 μg/L)자격HK-2세포24 h후,용RT-PCR방법검측세포중NF-kB mRNA급α평활기기동단백(α-SMA)mRNA표체,용형광광보법분석α-SMA단백적표체,병이도치상차현미경관찰세포전분화과정적형태변화.결과 TGF-β1유도24 h후,HK-2세포중NF-κB mRNA적표체현저상조,위공백대조조적8배이상(P<0.01).NF-kB반의과핵감산도입세포후,가현저억제TGF-β1유도적HK-2세포적NF-κB mRNA표체,비TGF-β1조감소75%(P<0.05),동시,α-SMA mRNA화단백표체역교TGF-β1조균명현하조(P<0.05).결론 NF-κB반의과핵감산가억제TGF-β1유도신소관상피세포NF-kB적표체,억제신소관상피세포전분화,가능유리우신간질섬유화적방치.
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-indueed epithelial mesenchymal transition was inhibited by NF-kB AS-ODN and the NF-KB 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.