Department of Hematology, The Second Hospital of Hebei Medical University, Hebei Key Laboratory of Hematology, No. 215, Hepingxi Road, Shijiazhuang, 050000, Hebei, China.
J Physiol Biochem. 2021 Feb;77(1):85-92. doi: 10.1007/s13105-021-00785-1. Epub 2021 Jan 30.
This study aimed to investigate the mechanism of type I interferon (IFN) in aggravating sepsis in bacterial infection, focusing on the roles of Caspase-11 (Casp11) and Gasdermin D (Gsdmd) in this process. Type I interferons, including IFNα and IFNβ, were used to treat peritoneal macrophage harvested from wild-type or IFNα/βR1 knockout (KO) mice, of which the levels of Casp11 and Gsdmd were monitored using real-time polymerase chain reaction (RT-PCR) and Western blot, the exposure to phosphatidylserine was monitored by flow cytometry, and tissue factor (TF) activation was assessed by RT-PCR and TF chromogenic assay. Endotoxemia in wild-type mice led to upregulation of Casp11 and Gsdmd in myeloid cells, which in contrast was attenuated in IFNα/βR1 KO mice. IFNα or IFNβ treatment led to dose-dependent upregulation of Casp11 and Gsdmd in peritoneal macrophages harvested from wild-type mice, but induced negligible changes in IFNα/βR1 KO mice. Type I IFN promoted phosphatidylserine exposure in peritoneal macrophage from wild-type mice but not IFNα/βR1 KO mice. Type I IFN induced insignificant changes of TF expression levels in both wild-type mice and IFNα/βR1 KO mice, but the TF activity was markedly increased in wild-type mice after type I IFN treatment. Our data suggested that the upregulation of Casp11 and Gsdmd in myeloid cells and macrophages induced by endotoxemia was reliant on the expression of IFNα/βR1. IFNα or IFNβ treatment efficiently upregulated Casp11 and Gsdmd, phosphatidylserine exposure, and TF activity of macrophages. Therefore, type I IFN could aggravate sepsis through upregulating Casp11 and Gsdmd.
本研究旨在探讨 I 型干扰素（IFN）在加重细菌感染性败血症中的作用机制，重点研究 Caspase-11（Casp11）和 Gasdermin D（Gsdmd）在这一过程中的作用。采用 IFNα 和 IFNβ 处理来自野生型或 IFNα/βR1 敲除（KO）小鼠的腹腔巨噬细胞，采用实时聚合酶链反应（RT-PCR）和 Western blot 检测 Casp11 和 Gsdmd 的水平，采用流式细胞术检测磷脂酰丝氨酸的暴露，采用 RT-PCR 和 TF 显色测定法评估组织因子（TF）的激活。野生型小鼠内毒素血症导致髓系细胞中 Casp11 和 Gsdmd 的上调，而 IFNα/βR1 KO 小鼠则减弱。IFNα 或 IFNβ 处理导致来自野生型小鼠的腹腔巨噬细胞中 Casp11 和 Gsdmd 的剂量依赖性上调，但在 IFNα/βR1 KO 小鼠中诱导的变化可忽略不计。I 型 IFN 促进了来自野生型小鼠的腹腔巨噬细胞中磷脂酰丝氨酸的暴露，但在 IFNα/βR1 KO 小鼠中没有诱导明显的变化。I 型 IFN 在野生型小鼠和 IFNα/βR1 KO 小鼠中均未引起 TF 表达水平的显著变化，但在 I 型 IFN 处理后，野生型小鼠中的 TF 活性显著增加。我们的数据表明，内毒素血症诱导的髓系细胞和巨噬细胞中 Casp11 和 Gsdmd 的上调依赖于 IFNα/βR1 的表达。IFNα 或 IFNβ 处理有效地上调了 Casp11 和 Gsdmd、磷脂酰丝氨酸的暴露和巨噬细胞的 TF 活性。因此，I 型 IFN 可通过上调 Casp11 和 Gsdmd 加重败血症。
