Department of Respiratory Medicine, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China.
Int Immunopharmacol. 2012 Apr;12(4):588-93. doi: 10.1016/j.intimp.2012.02.007. Epub 2012 Feb 29.
This study investigated whether aldose reductase (AR) inhibition affects interleukin (IL)-13-induced mucus production in the human bronchial epithelial cell line-16 (HBE16) cells. The HBE16 cells were cultured with AR inhibitors (zopolrestat) or were transfected with an AR small interfering (si)RNA. Subsequently, the cells were stimulated with 10 ng/ml IL-13 for 2h. The levels of mucin (MUC)5AC mRNA and protein were measured by using RT-PCR or ELISA. Intracellular reactive oxygen species (ROS) were measured fluorimetrically with the CM-H2DCFDA probe. Western blotting was performed to determine the levels of AR, phosphorylated signal transducer and activator of transcription 6 (p-STAT6) and phosphorylated Janus kinase 2 (p-JAK2). The results show that treatment with zopolrestat or transfection with AR siRNA significantly suppressed IL-13-stimulated MUC5AC mRNA and protein in the HBE16 cells (P<0.05). AR inhibition could suppress IL-13-induced ROS generation, the phosphorylation of JAK2/STAT6 pathway and the activation of nuclear factor (NF)-kappa B, thereby decreasing mucus production in vitro (all P<0.05). Therefore, the inhibition of AR could be a therapeutic target for mucus hypersecretion in chronic inflammation lung disease.
本研究旨在探讨醛糖还原酶(AR)抑制剂是否会影响人支气管上皮细胞系-16(HBE16)细胞中白细胞介素(IL)-13 诱导的粘蛋白产生。将 HBE16 细胞与 AR 抑制剂(唑泊司他)共培养或用 AR 小干扰(si)RNA 转染。随后,用 10ng/ml IL-13 刺激细胞 2h。通过 RT-PCR 或 ELISA 测定粘蛋白(MUC)5AC mRNA 和蛋白的水平。用 CM-H2DCFDA 探针通过荧光法测定细胞内活性氧(ROS)的水平。通过 Western blot 测定 AR、磷酸化信号转导和转录激活因子 6(p-STAT6)和磷酸化 Janus 激酶 2(p-JAK2)的水平。结果表明,唑泊司他处理或 AR siRNA 转染显著抑制了 HBE16 细胞中 IL-13 刺激的 MUC5AC mRNA 和蛋白(P<0.05)。AR 抑制可抑制 IL-13 诱导的 ROS 生成、JAK2/STAT6 通路的磷酸化和核因子(NF)-kappa B 的激活,从而减少体外粘液产生(均 P<0.05)。因此,AR 的抑制可能是慢性炎症性肺部疾病中粘液过度分泌的治疗靶点。