Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis

作者：Huiying Lu, Jian Lin, Chunjin Xu, Mingming Sun, Keqiang Zuo, Xiaoping Zhang, Mingsong Li, Hailiang Huang, Zhong Li, Wei Wu, Baisui Feng, Zhanju Liu · 发表于：Clinical and Translational Medicine · 年份：2021 · DOI：10.1002/ctm2.334 · 被引用次数：115 · 研究领域：Sirtuins and Resveratrol in Medicine、Neutrophil, Myeloperoxidase and Oxidative Mechanisms、Immune cells in cancer

BACKGROUND: Cyclosporine A (CsA) is routinely used to treat patients with steroid-refractory acute severe ulcerative colitis (ASUC). Here, we studied the underlying mechanisms of CsA-mediated alleviation in ASUC patients. METHODS: Neutrophil functions including expression of cytokines, apoptosis, and migration were measured by qRT-PCR, flow cytometry, and Transwell assay. Dynamic changes of glycolysis and tricarboxylic acid (TCA) cycle were measured by a Seahorse extracellular flux analyzer. Gene differences were determined and verified by RNA sequencing, qRT-PCR, and Western blotting. Small interfering RNA and inhibitors were used to knock down Sirtuin 6 (SIRT6) in HL-60 cells and block expression of SIRT6, hypoxia-inducible factor-1α (HIF-1α), and pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4) in neutrophils. RESULTS: We found that HIF-1α expression and glycolysis significantly increased, while the release of IL-8, myeloperoxidase (MPO) and reactive oxygen species (ROS), the apoptosis, and ability of migration markedly decreased in neutrophils of ASUC patients who responded to CsA (Response group) compared with those who did not respond to CsA (Nonresponse group). We also observed that CsA-induced functional alternation of neutrophils was initiated through suppressing SIRT6 expression, which is responsible for expression of the downstream signaling molecules (e.g., HIF-1α, PFKFB3) and PDK4 ubiquitination, leading to fueling neutrophil glycolysis and TCA cycle. Fur...