Huang Xiao, Han Shasha, Liu Xiangyong, Wang Tao, Xu Haixiao, Xia Bingyan, Kong Guiqing, Li Jiankui, Zhu Weiwei, Hu Haoran, Hao Dong, Wang Xiaozhi
Department of Respiratory Medicine and Intensive Care Unit, Binzhou Medical University Hospital, Binzhou, Shandong 256603, P.R. China.
Department of Cell Biology, Binzhou Medical University, Yantai, Shandong 264003, P.R. China.
Exp Ther Med. 2020 Feb;19(2):913-922. doi: 10.3892/etm.2019.8285. Epub 2019 Dec 5.
Sepsis commonly progresses to disseminated intravascular coagulation and induces the activation of heparanase (HPA) and the shedding of endothelial glycocalyx constituents, including syndecan-1 (SDC-1) and heparan sulphate (HS). However, the degradation of glycocalyx and its association with coagulation disorders remains undetermined. The present study aimed to evaluate the effect of unfractionated heparin (UFH) and N-acetylheparin (NAH), which is a non-anticoagulant heparin derivative, on endothelial glycocalyx and coagulation function in a lipopolysaccharide (LPS)-induced sepsis rat model, and to compare the differences observed in coagulation function between UFH and NAH. Experimental rats were randomly assigned to four groups: Control; LPS; UFH + LPS; and NAH + LPS. Rats were administered UFH or NAH and subsequently, ~1 min later, administered LPS (10 mg/kg; intravenous). The blood and lung tissues of rats were collected 0.5, 2 and 6 h after LPS injection, and were used for subsequent analysis. The results demonstrated that HPA activity and SDC-1 and HS levels increased, and this increase was associated with inflammatory cytokines and coagulation/fibrinolysis markers in the sepsis rat model. Histopathological examination was performed, and the lung injury score and lung wet/dry ratio indicated that UFH and NAH also significantly improved lung tissue injury. The results of the ELISA analysis demonstrated that UFH and NAH treatment: i) significantly decreased the levels of inflammatory cytokines including tumor necrosis factor-α and interleukin-6; ii) inhibited HPA activity and protected the integrity of the glycocalyx, which was identified by decreased HS and SDC-1 levels; and iii) decreased the levels of prothrombin fragment 1+2, thrombin-antithrombin complex, and plasminogen activator inhibitor-1 and increased the levels of fibrinogen and antithrombin-III. Preconditioning with UFH decreased the plasma activated partial thromboplastin time. These results indicated that UFH and NAH may alleviate sepsis-induced coagulopathy, and this effect may have been due to an inhibition of HPA activity and decrease in the shedding of the endothelial glycocalyx.
脓毒症通常会进展为弥散性血管内凝血,并诱导乙酰肝素酶(HPA)的激活以及内皮糖萼成分的脱落,包括多配体蛋白聚糖-1(SDC-1)和硫酸乙酰肝素(HS)。然而,糖萼的降解及其与凝血障碍的关联仍未明确。本研究旨在评估普通肝素(UFH)和N-乙酰肝素(NAH,一种非抗凝肝素衍生物)对脂多糖(LPS)诱导的脓毒症大鼠模型中内皮糖萼和凝血功能的影响,并比较UFH和NAH在凝血功能方面观察到的差异。将实验大鼠随机分为四组:对照组;LPS组;UFH + LPS组;以及NAH + LPS组。给大鼠注射UFH或NAH,随后,约1分钟后,注射LPS(10 mg/kg;静脉注射)。在注射LPS后0.5、2和6小时收集大鼠的血液和肺组织,用于后续分析。结果表明,在脓毒症大鼠模型中,HPA活性以及SDC-1和HS水平升高,且这种升高与炎性细胞因子和凝血/纤溶标志物相关。进行了组织病理学检查,肺损伤评分和肺湿/干比表明UFH和NAH也显著改善了肺组织损伤。ELISA分析结果表明,UFH和NAH治疗:i)显著降低了包括肿瘤坏死因子-α和白细胞介素-6在内的炎性细胞因子水平;ii)抑制了HPA活性并保护了糖萼的完整性,这通过HS和SDC-1水平降低得以证实;iii)降低了凝血酶原片段1+2、凝血酶-抗凝血酶复合物以及纤溶酶原激活物抑制剂-1的水平,并提高了纤维蛋白原和抗凝血酶-III的水平。用UFH预处理可缩短血浆活化部分凝血活酶时间。这些结果表明,UFH和NAH可能减轻脓毒症诱导的凝血病,且这种作用可能归因于对HPA活性的抑制以及内皮糖萼脱落的减少。