Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Korea.
Department of Stem cell and Regenerative Biotechnology, Konkuk University, Seoul 05029, Korea.
Int J Mol Sci. 2019 Apr 20;20(8):1949. doi: 10.3390/ijms20081949.
Interleukin (IL)-32θ, a newly identified IL-32 isoform, has been reported to exert pro-inflammatory effects through the association with protein kinase C delta (PKCδ). In this study, we further examined the effects of IL-32θ on IL-13 and IL-13Rα2 expression and the related mechanism in THP-1 cells. Upon stimulating IL-32θ-expressing and non-expressing cells with phorbol 12-myristate 13-acetate (PMA), the previous microarray analysis showed that IL-13Rα2 and IL-13 mRNA expression were significantly decreased by IL-32θ. The protein expression of these factors was also confirmed to be down-regulated. The nuclear translocation of transcription factors STAT3 and STAT6, which are necessary for IL-13Rα2 and IL-13 promoter activities, was suppressed by IL-32θ. Additionally, a direct association was found between IL-32θ, PKCδ, and signal transducer and activator of transcription 3 (STAT3), but not STAT6, revealing that IL-32θ might act mainly through STAT3 and indirectly affect STAT6. Moreover, the interaction of IL-32θ with STAT3 requires PKCδ, since blocking PKCδ activity eliminated the interaction and consequently limited the inhibitory effect of IL-32θ on STAT3 activity. Interfering with STAT3 or STAT6 binding by decoy oligodeoxynucleotides (ODNs) identified that IL-32θ had additive effects with the STAT3 decoy ODN to suppress IL-13 and IL-13Rα2 mRNA expression. Taken together, our data demonstrate the intracellular interaction of IL-32θ, PKCδ, and STAT3 to regulate IL-13 and IL-13Rα2 synthesis, supporting the role of IL-32θ as an inflammatory modulator.
白细胞介素(IL)-32θ 是一种新发现的 IL-32 同工型,据报道通过与蛋白激酶 C 三角洲(PKCδ)的结合发挥促炎作用。在这项研究中,我们进一步研究了 IL-32θ 对 THP-1 细胞中 IL-13 和 IL-13Rα2 表达的影响及相关机制。用佛波醇 12-肉豆蔻酸 13-乙酸酯(PMA)刺激表达和不表达 IL-32θ 的细胞后,先前的微阵列分析显示,IL-32θ 显著降低了 IL-13Rα2 和 IL-13 mRNA 的表达。这些因子的蛋白表达也被证实下调。转录因子 STAT3 和 STAT6 的核易位被 IL-32θ 抑制,这些因子对于 IL-13Rα2 和 IL-13 启动子活性是必需的。此外,发现 IL-32θ 与 PKCδ 和信号转导和转录激活因子 3(STAT3)之间存在直接关联,但与 STAT6 无关,表明 IL-32θ 可能主要通过 STAT3 发挥作用,并间接影响 STAT6。此外,IL-32θ 与 STAT3 的相互作用需要 PKCδ,因为阻断 PKCδ 活性消除了相互作用,从而限制了 IL-32θ 对 STAT3 活性的抑制作用。通过诱饵寡脱氧核苷酸(ODN)干扰 STAT3 或 STAT6 的结合,确定了 IL-32θ 与 STAT3 诱饵 ODN 具有协同作用,可抑制 IL-13 和 IL-13Rα2 mRNA 的表达。总之,我们的数据表明 IL-32θ、PKCδ 和 STAT3 之间的细胞内相互作用调节 IL-13 和 IL-13Rα2 的合成,支持 IL-32θ 作为炎症调节剂的作用。