Zeng Xiaoyan, Li Jiuxi, Pei Liyuan, Yang Yaping, Chen Ya, Wang Xuejing, Zhang Ting, Zhou Ting
Department of Clinical Chinese Pharmacy, College of Pharmacy Hunan University of Chinese Medicine Changsha Hunan China.
College of Acupuncture, Massage and Rehabilitation Hunan University of Chinese Medicine Changsha Hunan China.
Pulm Circ. 2024 Oct 9;14(4):e12454. doi: 10.1002/pul2.12454. eCollection 2024 Oct.
This research aims to investigate the impact of Didang decoction (DD) on the formation of neutrophil extracellular traps (NETs) and cancer-associated thrombosis in lung cancer. BALB/c nude mice were used to establish xenograft models for inducing deep vein thrombosis. Tumor growth and thrombus length were assessed. The impact of DD on NET generation was analyzed using enzyme-linked immunosorbent assay, immunofluorescence staining, quantitative real-time PCR, and western blot analysis, both in vivo and in vitro. CI-amidine, a PAD4 inhibitor, was employed to evaluate the role of PAD4 in the generation of NETs. In vivo studies demonstrated that treatment with DD reduced tumor growth, inhibited thrombus formation, and decreased the levels of NET markers in the serum, tumor tissues, neutrophils, and thrombus tissues of mice. Additional data indicated that DD could suppress neutrophil counts, the release of tissue factor (TF), and the activation of thrombin-activated platelets, all of which contributed to increased formation of NETs in mouse models. In vitro, following incubation with conditioned medium (CM) derived from Lewis lung carcinoma cells, the expression of NET markers in neutrophils was significantly elevated, and an extracellular fibrous network structure was observed. Nevertheless, these NET-associated changes were partially counteracted by DD. Additionally, CI-amidine reduced the expression of NET markers in CM-treated neutrophils, consistent with the effects of DD. Collectively, DD inhibits cancer-associated thrombosis in lung cancer by decreasing PAD4-dependent NET formation through the regulation of TF-mediated thrombin-platelet activation. This presents a promising therapeutic strategy for preventing and treating venous thromboembolism in lung cancer.
本研究旨在探讨抵当汤（DD）对肺癌中性粒细胞胞外诱捕网（NETs）形成及癌症相关血栓形成的影响。采用BALB/c裸鼠建立诱导深静脉血栓形成的异种移植模型。评估肿瘤生长和血栓长度。通过酶联免疫吸附测定、免疫荧光染色、定量实时聚合酶链反应和蛋白质免疫印迹分析，在体内和体外分析DD对NET生成的影响。使用PAD4抑制剂CI-脒来评估PAD4在NET生成中的作用。体内研究表明，DD治疗可减少肿瘤生长、抑制血栓形成，并降低小鼠血清、肿瘤组织、中性粒细胞和血栓组织中NET标志物的水平。其他数据表明，DD可抑制中性粒细胞计数、组织因子（TF）的释放以及凝血酶激活的血小板的活化，所有这些因素都导致小鼠模型中NET形成增加。在体外，与Lewis肺癌细胞来源的条件培养基（CM）孵育后，中性粒细胞中NET标志物的表达显著升高，并观察到细胞外纤维网络结构。然而，这些与NET相关 的变化被DD部分抵消。此外，CI-脒降低了CM处理的中性粒细胞中NET标志物的表达，这与DD的作用一致。总体而言，DD通过调节TF介导的凝血酶-血小板活化，减少PAD4依赖性NET形成，从而抑制肺癌中的癌症相关血栓形成。这为预防和治疗肺癌中的静脉血栓栓塞提供了一种有前景的治疗策略。
