Mishina Kei, Shinkai Masaharu, Shimokawaji Tadasuke, Nagashima Akimichi, Hashimoto Yusuke, Inoue Yoriko, Inayama Yoshiaki, Rubin Bruce K, Ishigatsubo Yoshiaki, Kaneko Takeshi
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Int Immunopharmacol. 2015 Dec;29(2):448-453. doi: 10.1016/j.intimp.2015.10.016. Epub 2015 Oct 24.
Mucus hypersecretion and goblet cell hyperplasia are common features that characterize asthma. IL-13 increases mucin (MUC) 5AC, the major component of airway mucus, in airway epithelial cells. According to the literature, IL-13 receptor activation leads to STAT6 activation and consequent induction of chloride channel accessory 1 (CLCA1) gene expression, associated with the induction of MUC5AC. Heme oxygenase-1 (HO-1) is an enzyme that catalyzes oxidation of heme to biliverdin, and has anti-inflammatory and anti-oxidant properties. We examined the effects of HO-1 on mucin production and goblet cell hyperplasia induced by IL-13. Moreover, we assessed the cell signaling intermediates that appear to be responsible for mucin production. Normal human bronchial epithelial (NHBE) cells were grown at air liquid interface (ALI) in the presence or absence of IL-13 and hemin, a HO-1 inducer, for 14 days. Protein concentration was analyzed using ELISA, and mRNA expression was examined by real-time PCR. Histochemical analysis was performed using HE staining, andWestern blotting was performed to evaluate signaling transduction pathway. Hemin (4 μM) significantly increased HO-1 protein expression (p b 0.01) and HO-1 mRNA expression (p b 0.001). IL-13 significantly increased goblet cells, MUC5AC protein secretion (p b 0.01) and MUC5AC mRNA (p b 0.001), and these were decreased by hemin by way of HO-1. Tin protoporphyrin (SnPP)-IX, a HO-1 inhibitor, blocked the effect of hemin restoring MUC5AC protein secretion (p b 0.05) and goblet cell hyperplasia. Hemin decreased the expression of CLCA1 mRNA (p b 0.05) and it was reversed by SnPP-IX, but could not suppress IL-13-induced phosphorylation of STAT6 or SAM pointed domain-containing ETS transcription factor (SPDEF) and Forkhead box A2 (FOXA2) mRNA expression. In summary, HO-1 overexpression suppressed IL-13-induced goblet cell hyperplasia and MUC5AC production, and involvement of CLCA1 in the mechanism was suggested.
黏液高分泌和杯状细胞增生是哮喘的常见特征。白细胞介素-13（IL-13）可增加气道上皮细胞中气道黏液的主要成分黏蛋白（MUC）5AC。根据文献报道，IL-13受体激活会导致信号转导和转录激活因子6（STAT6）激活，进而诱导与MUC5AC诱导相关的氯通道辅助蛋白1（CLCA1）基因表达。血红素加氧酶-1（HO-1）是一种催化血红素氧化为胆绿素的酶，具有抗炎和抗氧化特性。我们研究了HO-1对IL-13诱导的黏蛋白产生和杯状细胞增生的影响。此外，我们评估了似乎与黏蛋白产生有关的细胞信号中间体。正常人支气管上皮（NHBE）细胞在气液界面（ALI）培养，分别在有或无IL-13和HO-1诱导剂血红素的情况下培养14天。使用酶联免疫吸附测定（ELISA）分析蛋白质浓度，通过实时聚合酶链反应（PCR）检测mRNA表达。使用苏木精-伊红（HE）染色进行组织化学分析，并进行蛋白质印迹法以评估信号转导途径。血红素（4μM）显著增加HO-1蛋白表达（p<0.01）和HO-1 mRNA表达（p<0.001）。IL-13显著增加杯状细胞、MUC5AC蛋白分泌（p<0.01）和MUC5AC mRNA（p<0.001），而血红素通过HO-1降低了这些指标。HO-1抑制剂锡原卟啉（SnPP）-IX阻断了血红素恢复MUC5AC蛋白分泌（p<0.05）和杯状细胞增生的作用。血红素降低了CLCA1 mRNA的表达（p<0.05），且被SnPP-IX逆转，但不能抑制IL-13诱导的STAT6磷酸化或含SAM结构域的ETS转录因子（SPDEF）以及叉头框A2（FOXA2）mRNA表达。总之，HO-1过表达抑制了IL-13诱导的杯状细胞增生和MUC5AC产生，并提示CLCA1参与了该机制。
