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目的:探讨Akt信号通路在肾小管上皮细胞(NRK-52E)分泌转化生长因子β1(TGF-β1)和核因子-κB(NF-κB)表达中的作用。方法:体外培养NRK-52E细胞,分别加入不同浓度(5,15,30g/L)白蛋白和/或Akt抑制剂Ly294002。应用RT-PCR方法测定TGF-β1mRNA表达,Western blot测定Akt和TGF-β1蛋白的表达。应用凝胶电泳迁移率(EMSA)检测NF-κB活化。两两比较用t检验。结果:白蛋白呈浓度依赖性上调肾小管上皮细胞TGF-β1mRNA表达增加。TGF-β1蛋白表达也增加。白蛋白激活NF-κB活性,增加磷酸化Akt表达。Ly294002能够抑制白蛋白引起的NF-κB活性以及TGF-β1的表达。NF-κB活化与TGF-β1表达呈显著正相关(r=0.61,P<0.05)。结论:白蛋白刺激肾小管上皮细胞分泌TGF-β1和NF-κB的活性增加,其机制部分依赖于Akt的磷酸化作用。
AIM: To investigate the role of Akt pathway in the expression of transforming growth factor-β1 (TGF-β1) and nuclear factor-κB (NF-κB) in renal tubular epithelial cells (NRK-52E). Methods: NRK-52E cells were cultured in vitro, and different concentrations (5, 15 and 30 g / L) of albumin and / or Akt inhibitor Ly294002 were added respectively. The expression of TGF-β1 mRNA was detected by RT-PCR and the expression of Akt and TGF-β1 by Western blot. NF-κB activation was detected by gel electrophoretic mobility shift assay (EMSA). Pairwise comparisons using t test. Results: Albumin increased the expression of TGF-β1 mRNA in renal tubular epithelial cells in a concentration-dependent manner. TGF-β1 protein expression is also increased. Albumin activates NF-κB activity and increases phosphorylated Akt expression. Ly294002 inhibited albumin-induced NF-κB activity and TGF-β1 expression. There was a significant positive correlation between NF-κB activation and TGF-β1 expression (r = 0.61, P <0.05). Conclusion: Albumin stimulates the secretion of TGF-β1 and NF-κB in renal tubular epithelial cells. The mechanism is partially dependent on the phosphorylation of Akt.