圣草次苷调节SphK1/S1P通路对脂多糖诱导的脓毒症内皮细胞炎症损伤的影响  

The effect of eriocitrin on lipopolysaccharide-induced inflammation of endothelial cells in sepsis by regulating SphK1/S1P pathway

作　　者：张开亚 徐瑶瑶 杨琴[1] 李夏 Zhang Kaiya;Xu Yaoyao;Yang Qin;Li Xia(Department of Critical Medicine,Wenzhou Hospital of Traditional Chinese Medicine,Zhejiang Chinese Medicine University,Wenzhou 325000,China)

机构地区：[1]浙江中医药大学附属温州市中医院重症医学科,浙江温州325000

出　　处：《中国急救医学》2025年第2期157-162,共6页Chinese Journal of Critical Care Medicine

摘　　要：目的研究圣草次苷对脂多糖(LPS)诱导的脓毒症内皮细胞炎症损伤的修复作用,并基于鞘氨醇激酶1(SphK1)/1-磷酸鞘氨醇(S1P)通路探讨圣草次苷对脓毒症内皮细胞的作用机制。方法人脐静脉内皮细胞(HUVEC)分为空白对照组、模型组、圣草次苷低、中、高剂量组及阳性组,模型组、圣草次苷低、中、高剂量组及阳性组分别加入终浓度为100 ng/mL的脂多糖(LPS),继续培养24 h,制备脓毒症模型。圣草次苷低、中、高剂量组分别加入终浓度为100、200、400μg/mL的圣草次苷,阳性组加入终浓度为10μmol/L的新补骨脂异黄酮,空白对照组及模型组加入等量培养液。使用酶联免疫吸附法(ELISA)试剂盒测定HUVEC上清一氧化氮(NO)、白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、巨噬细胞趋化蛋白-1(MCP-1)、干扰素(IFN)-γ、IL-8和细胞髓过氧化物酶(MPO)、超氧化物歧化酶(SOD)、丙二醛(MDA)、C反应蛋白(CRP)、乳酸脱氢酶(LDH),使用跨膜电阻仪测定跨膜电阻(TER),荧光标记法测定HUVEC通透性,使用凋亡试剂盒测定HUVEC凋亡率,实时定量聚合酶链式反应(PCR)和免疫印记法(Western blot)分别测定各组HUVEC的SphK1及S1P信使核糖核酸(mRNA)及蛋白水平。结果与空白对照组比较,模型组HUVEC上清NO、IL-6、TNF-α、GM-CSF、MCP-1、IFN-γ及IL-8水平,MPO、MDA、CRP、LDH水平,HUVEC通透性及凋亡率,SphK1及S1P mRNA及蛋白水平升高,SOD及TER降低,差异有统计学意义(P<0.05);与模型组比较,圣草次苷低、中、高剂量组及阳性组HUVEC上清NO、IL-6、TNF-α、GM-CSF、MCP-1、IFN-γ及IL-8水平,MPO、MDA、CRP、LDH水平,HUVEC通透性及凋亡率,SphK1及S1P mRNA及蛋白水平降低,SOD及TER升高,差异有统计学意义(P<0.05)。结论圣草次苷能够显著抑制LPS诱导的脓毒症内皮细胞炎症反应,改善LPS诱导的脓毒症内皮细胞氧化损伤,修复脓毒症内皮细胞通透性,进�Objective To study the repair effect of eriocitrin on lipopolysaccharide(LPS)-induced inflammation of endothelial cells in sepsis,and to explore the mechanism of eriocitrin in endothelial cells in sepsis based on the sphingosine kinase 1/sphingosine-1-phosphate(SphK1/S1P)pathway.Methods Human umbilical vein endothelial cells(HUVEC)were divided into blank control group,model group,low-dose,medium-dose and high-dose eriocitrin groups and positive group.The model group,low-dose,medium-dose and high-dose eriocitrin groups and positive group were added with final concentration of 100 ng/mL LPS for 24 h,and the sepsis model was prepared.The low-dose,medium-dose and high-dose groups were added with the final concentration of 100,200 and 400μg/mL eriocitrin,the positive group was added with the final concentration of 10μmol/L of new psoralen isoflavones,the blank control group and the model group were added with the culture medium.Levels of nitric oxide(NO),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),granulocyte-macrophage colony-stimulating factor(GM-CSF),monocyte chemoattractant protein-1(MCP-1),interferon-γ(IFN-γ),interleukin-8(IL-8)in HUVEC supernatant and cellular myeloperoxidase(MPO),superoxide disumutase(SOD),malondialdehyde(MDA),C-reactive protein(CRP)and lactate dehydrogenase(LDH)were determined by enzyme-linked immunosorbent assay(ELISA)kit,transmembrane resistance(TER)was determined by transmembrane resistance meter,and HUVEC permeability was determined by fluorescence labeling method.The apoptotic rate of HUVEC was determined by apoptosis kit.The mRNA and protein levels of SphK1 and S1P of HUVEC in different groups were determined by real-time quantitative polymerase chain reaction(PCR)and Western blot,respectively.Results Compared with blank control group,HUVEC supernatant levels of NO,IL-6,TNF-α,GM-CSF,MCP-1,IFN-γand IL-8,MPO,MDA,CRP and LDH,HUVEC permeability and apoptosis rate,the mRNA and protein levels of SphK1 and S1P were increased in model group,SOD and TER were decreased,the differenc

关 键 词：圣草次苷 SphK1/S1P通路 脓毒症 炎症损伤 

分 类 号：R459.7[医药卫生—急诊医学]