Zhong Xin, Zhao Xueying, Zhang Liyuan, Liu Ning, Shi Sa, Wang Yuwen
Department of Pathophysiology, Harbin Medical University, Harbin, 150081, China.
Department of Clinical Laboratory, The second Affiliated Hospital of Harbin Medical University, Harbin, 150081, China.
Biochem Biophys Res Commun. 2022 May 28;606:1-9. doi: 10.1016/j.bbrc.2022.03.072. Epub 2022 Mar 18.
Hydrogen sulfide (HS) prevents endothelial cells injury. However, the complicated mechanism of sodium hydrosulfide (NaHS, a donor that produces HS) which inhibits the endothelial cells injury which correlated the activation of neutrophil in the type 1 diabetes mellitus (T1DM) rats has not been previously investigated.
In the experiment, the T1DM animal model was established, the IL-1β, IL-8 were determined by western blotting and ELISA, the expressions of the Bax and Bcl-2 of endothelial cells and the CXCR2, CSE, phosphor-IκBα and NF-kB of neutrophils were measured by western blotting. Additionally, the concentration of serum dsDNA was tested by PicoGreen commercial Kits, changes in the HS concentration of neutrophils were determined by Multiskan spectrum microphate spectrophotometer, the cellular ROS levels of neutrophils were detected by DCFH-DA staining and flow cytometry. The IL-1β, IL-8 concentration and expression increased, the endothelial cells injury which stimulated by high glucose and the concentration of dsDNA in serum increased, the expression of CXCR2, phosphor-IκBα and NF-kB increased while the expression of CSE and concentration of HS decreased in neutrophils in the T1DM group compared to the control group. NaHS significantly inhibited the injury of endothelial cell, the production of ROS in neutrophils, reversed the expressions of CXCR2, CSE, phosphor-IκBα and NF-κB and decreased concentration of dsDNA in serum which were caused by T1DM.
Our results demonstrated that the donor of HS inhibits endothelial cells injury and neutrophils activation via the IL-8/CXCR2/ROS/NF-κB axis in T1DM rat.
硫化氢（HS）可预防内皮细胞损伤。然而，硫氢化钠（NaHS，一种产生HS的供体）抑制内皮细胞损伤的复杂机制，该机制与1型糖尿病（T1DM）大鼠中性粒细胞的激活相关，此前尚未得到研究。
在实验中，建立T1DM动物模型，通过蛋白质免疫印迹法和酶联免疫吸附测定法测定白细胞介素-1β（IL-1β）、白细胞介素-8（IL-8），通过蛋白质免疫印迹法测量内皮细胞中Bax和Bcl-2的表达以及中性粒细胞中CXC趋化因子受体2（CXCR2）、胱硫醚-γ-裂解酶（CSE）、磷酸化核因子κB抑制蛋白α（phosphor-IκBα）和核因子κB（NF-κB）的表达。此外，通过PicoGreen商用试剂盒检测血清双链DNA（dsDNA）浓度，用多功能酶标仪测定中性粒细胞中HS浓度的变化，通过2′,7′-二氯二氢荧光素二乙酸酯（DCFH-DA）染色和流式细胞术检测中性粒细胞的细胞活性氧（ROS）水平。与对照组相比，T1DM组中IL-1β、IL-8浓度和表达增加，高糖刺激引起的内皮细胞损伤和血清中dsDNA浓度增加，中性粒细胞中CXCR2、phosphor-IκBα和NF-κB的表达增加，而CSE的表达和HS的浓度降低。NaHS显著抑制内皮细胞损伤、中性粒细胞中ROS的产生，逆转了由T1DM引起的CXCR2、CSE、phosphor-IκBα和NF-κB的表达，并降低了血清中dsDNA的浓度。
我们的结果表明，HS供体通过T1DM大鼠中的IL-8/CXCR2/ROS/NF-κB轴抑制内皮细胞损伤和中性粒细胞激活。
