Sodium trisulfide (Na2S3) releases hydrogen polysulfide (H2Sn) and is useful for the investigation of the effects of H2Sn on the cell functions. In the present study, we first examined the effects of Na2S3 on the gene expression of IEC-6 cells, a rat intestinal epithelial cell line. Microarray analysis and reverse transcription-polymerase chain reaction analysis revealed that Na2S3 increased the gene expression of early growth response 1 (EGR1) and Kruppel-like transcription factor 4 (KLF4). It was interesting that U0126, an inhibitor of the activation of extracellular signal-regulated kinase 1 (ERK1), ERK2, and ERK5, inhibited the Na2S3-induced gene expression of EGR1 and KLF4. Na2S3 activated ERK1 and ERK2 (ERK1/2) within 15 min. In addition to ERK1/2, Na2S3 activated ERK5. We noticed that the electrophoretic mobility of ERK5 was decreased after Na2S3 treatment. Phos-tag analysis and in vitro dephosphorylation of the cell extracts indicated that the gel-shift of ERK5 was due to its phosphorylation. The gel-shift of ERK5 was inhibited completely by both U0126 and ERK5-IN-1, a specific inhibitor of ERK5. From these results, we concluded that the gel-shift of ERK5 was induced through autophosphorylation by activated ERK5 after Na2S3 treatment. The present study suggested that H2Sn affected various functions of intestinal epithelial cells through the activation of the ERK1/2 and ERK5 pathways.

中文翻译：

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通过活化培养的肠上皮细胞中的ERK1 / 2和ERK5途径，多硫化物增加了EGR1和KLF4的表达。

三硫化钠（Na2S3）释放多硫化氢（H2Sn），可用于研究H2Sn对细胞功能的影响。在本研究中，我们首先检查了Na2S3对大鼠肠上皮细胞系IEC-6细胞的基因表达的影响。微阵列分析和逆转录-聚合酶链反应分析表明，Na2S3增加了早期生长反应1（EGR1）和Kruppel样转录因子4（KLF4）的基因表达。有趣的是，U0126是细胞外信号调节激酶1（ERK1），ERK2和ERK5激活的抑制剂，可抑制Na2S3诱导的EGR1和KLF4的基因表达。Na2S3在15分钟内激活了ERK1和ERK2（ERK1 / 2）。除了ERK1 / 2，Na2S3还激活了ERK5。我们注意到，Na2S3处理后ERK5的电泳迁移率降低了。细胞提取物的磷酸标签分析和体外去磷酸化表明ERK5的凝胶位移是由于其磷酸化引起的。U0126和ERK5的特异性抑制剂ERK5-IN-1均完全抑制ERK5的凝胶转移。从这些结果，我们得出结论，Na2S3处理后，活化的ERK5通过自磷酸化诱导ERK5的凝胶转变。本研究表明，H2Sn通过激活ERK1 / 2和ERK5途径影响肠道上皮细胞的各种功能。ERK5的特异性抑制剂。从这些结果，我们得出结论，Na2S3处理后，活化的ERK5通过自磷酸化诱导ERK5的凝胶迁移。本研究表明，H2Sn通过激活ERK1 / 2和ERK5途径影响肠道上皮细胞的各种功能。ERK5的特异性抑制剂。从这些结果，我们得出结论，Na2S3处理后，活化的ERK5通过自磷酸化诱导ERK5的凝胶迁移。本研究表明，H2Sn通过激活ERK1 / 2和ERK5途径影响肠道上皮细胞的各种功能。