Lan Hong-Tao, Zheng Ya-Ting, Tong Zhou-Jie, Zhang Cong, Cong Xiao-Yan, Wang Zhi-Hao
The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
Department of Geriatric Medicine, Qilu Hospital of Shandong University, Key Laboratory of Cardiovascular Proteomics of Shandong Province, Qilu Hospital of Shandong University, Jinan, China.
Evid Based Complement Alternat Med. 2022 Dec 21;2022:6606423. doi: 10.1155/2022/6606423. eCollection 2022.
We aimed to investigate the effects of the natural product humic acids (HA) on platelet activation and development of venous thromboembolism (VTE) in mice and further explore the relevant mechanism.
Eight-week C57BL/6 mice were randomly assigned to three groups: sham operation group ( = 7), VTE group ( = 8), and VTE + HA group ( = 10). Thrombi were harvested to hematoxylin-eosin staining to evaluate the thrombolysis and recanalization of the thrombus. In addition, flow cytometry was performed to detect the expression levels of protein disulfide isomerase on endothelial-derived exosomes and glycoprotein IIb/IIIa on the surface of the activated platelets surface in plasma. Furthermore, the protein expression level of glycoprotein IIb/IIIa in thrombus was determined by immunohistochemistry and immunofluorescence.
The length of thrombosis in the VTE + HA group was significantly shorter than that in the VTE group ( = 0.040). No significant differences were observed in thrombolysis and recanalization between the VTE + HA group and the VTE group ( > 0.05). The content of protein disulfide isomerase carried by endothelial-derived exosomes was significantly increased in the VTE group ( = 0.008) but significantly reduced by native humic acids ( = 0.012). Compared with the VTE group, the expression of glycoprotein IIb/IIIa on activated platelet surface in the VTE + HA group was significantly decreased ( = 0.002). The concentration of plasmatic P-selectin in the VTE group was significantly higher than that in the VTE + HA group ( < 0.001).
We demonstrate that HA possess a pharmacological property that decreases venous thrombus formation in mice. The underlying mechanism is that HA could inhibit the expression of glycoprotein IIb/IIIa on the activated platelets surface by suppressing endothelial-derived exosomes carrying on protein disulfide isomerase, thereby blocking platelet activation.
我们旨在研究天然产物腐殖酸（HA）对小鼠血小板活化及静脉血栓栓塞症（VTE）发展的影响，并进一步探究相关机制。
将8周龄的C57BL/6小鼠随机分为三组：假手术组（n = 7）、VTE组（n = 8）和VTE + HA组（n = 10）。采集血栓进行苏木精-伊红染色，以评估血栓的溶解和再通情况。此外，采用流式细胞术检测血浆中内皮细胞衍生外泌体上蛋白质二硫键异构酶的表达水平以及活化血小板表面糖蛋白IIb/IIIa的表达水平。此外，通过免疫组织化学和免疫荧光法测定血栓中糖蛋白IIb/IIIa的蛋白表达水平。
VTE + HA组的血栓长度明显短于VTE组（P = 0.040）。VTE + HA组与VTE组在血栓溶解和再通方面未观察到显著差异（P > 0.05）。VTE组中内皮细胞衍生外泌体携带的蛋白质二硫键异构酶含量显著增加（P = 0.008），但天然腐殖酸使其显著降低（P = 0.012）。与VTE组相比，VTE + HA组活化血小板表面糖蛋白IIb/IIIa的表达明显降低（P = 0.002）。VTE组血浆P-选择素浓度明显高于VTE + HA组（P < 0.001）。
我们证明HA具有降低小鼠静脉血栓形成的药理特性。潜在机制是HA可通过抑制携带蛋白质二硫键异构酶的内皮细胞衍生外泌体，从而抑制活化血小板表面糖蛋白IIb/IIIa的表达，进而阻断血小板活化。
