Departments of Otorhinolaryngology-Head and Neck Surgery, Inje University, College of Medicine, Pusan Paik Hospital, Busan, Korea.
Am J Rhinol Allergy. 2013 Jan;27(1):e26-31. doi: 10.2500/ajra.2013.27.3849.
Methotrexate (MTX) is a very effective treatment for chronic inflammatory diseases that induces apoptosis in nasal polyps (NPs). However, the precise apoptotic pathway in NPs remains unclear. The aim of this study was to identify the apoptotic signaling pathways activated by MTX in NPs.
NP tissues were organ cultured using an air-liquid interface method. Cultures were maintained in the presence or absence of MTX (10 or 100 μM) for 24 hours. To investigate apoptotic signaling in NPs, we performed reverse transcription-polymerase chain reaction and Western blotting.
MTX-treated NPs contained significantly increased amounts of the active forms of caspase 8, caspase 9, and caspase 3 and displayed increased cleavage of poly(ADP-ribose) polymerase. Expression of the proapoptotic molecules Bax and Bad at the mRNA and protein levels and of the activated molecules p-Bad and tBid was significantly higher in MTX-treated NPs than in nontreated NPs. In contrast, expression of the antiapoptotic molecule Bcl-2 at the mRNA and protein levels was significantly lower in MTX-treated NPs than in nontreated NPs. Expression of the phosphorylated forms of p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK) was significantly higher in MTX-treated NPs than in nontreated NPs. In contrast, expression of the phosphorylated form of Akt was significantly lower in MTX-treated NPs than in nontreated NPs.
MTX induces apoptosis in NPs via caspase cascades and both mitochondria-mediated and p38 MAPK/JNK pathways. We suggest that MTX can be used to treat NPs.
甲氨蝶呤(MTX)是一种非常有效的治疗慢性炎症性疾病的药物,可诱导鼻息肉(NPs)凋亡。然而,NPs 中确切的凋亡途径尚不清楚。本研究旨在确定 MTX 在 NPs 中激活的凋亡信号通路。
采用气液界面法对 NP 组织进行器官培养。在存在或不存在 MTX(10 或 100 μM)的情况下培养 24 小时。为了研究 NPs 中的凋亡信号,我们进行了逆转录-聚合酶链反应和 Western 印迹。
MTX 处理的 NPs 中含有明显增加的活性胱天蛋白酶 8、胱天蛋白酶 9 和胱天蛋白酶 3,并且多聚(ADP-核糖)聚合酶的切割增加。在 MTX 处理的 NPs 中,促凋亡分子 Bax 和 Bad 的 mRNA 和蛋白水平以及激活的分子 p-Bad 和 tBid 的表达明显高于未处理的 NPs。相比之下,MTX 处理的 NPs 中抗凋亡分子 Bcl-2 的 mRNA 和蛋白水平明显低于未处理的 NPs。MTX 处理的 NPs 中磷酸化形式的 p38 丝裂原活化蛋白激酶(MAPK)和 c-Jun N 端激酶(JNK)的表达明显高于未处理的 NPs。相比之下,MTX 处理的 NPs 中磷酸化形式的 Akt 的表达明显低于未处理的 NPs。
MTX 通过胱天蛋白酶级联和线粒体介导以及 p38 MAPK/JNK 途径诱导 NPs 凋亡。我们建议 MTX 可用于治疗 NPs。