Laboratory of Ethnopharmacology, Tissue-Orientated Property of Chinese Medicine Key Laboratory of Sichuan Province, West China School of Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Int J Mol Sci. 2023 Feb 19;24(4):4168. doi: 10.3390/ijms24044168.
Disseminated intravascular coagulation (DIC), which is closely related to platelet activation, is a key factor leading to high mortality in sepsis. The release of contents from plasma membrane rupture after platelet death further aggravates thrombosis. Nerve injury-induced protein 1 (NINJ1) is a cell membrane protein that mediates membrane disruption, a typical marker of cell death, through oligomerization. Nevertheless, whether NINJ1 is expressed in platelets and regulates the platelet function remains unclear. The aim of this study was to evaluate the expression of NINJ1 in human and murine platelets and elucidate the role of NINJ1 in platelets and septic DIC. In this study, NINJ1 blocking peptide (NINJ1) was used to verify the effect of NINJ1 on platelets in vitro and in vivo. Platelet αIIbβ3 and P-selectin were detected by flow cytometry. Platelet aggregation was measured by turbidimetry. Platelet adhesion, spreading and NINJ1 oligomerization were examined by immunofluorescence. Cecal perforation-induced sepsis and FeCl-induced thrombosis models were used to evaluate the role of NINJ1 in platelet, thrombus and DIC in vivo. We found that inhibition of NINJ1 alleviates platelet activation in vitro. The oligomerization of NINJ1 is verified in membrane-broken platelets, which is regulated by the PANoptosis pathway. In vivo studies demonstrate that inhibition of NINJ1 effectively reduces platelet activation and membrane disruption, thus suppressing platelet-cascade reaction and leading to anti-thrombosis and anti-DIC in sepsis. These data demonstrate that NINJ1 is critical in platelet activation and plasma membrane disruption, and inhibition of NINJ1 effectively reduces platelet-dependent thrombosis and DIC in sepsis. This is the first study to reveal the key role of NINJ1 in platelet and its related disorders.
弥散性血管内凝血（DIC）与血小板激活密切相关，是导致脓毒症高死亡率的关键因素。血小板死亡后，细胞膜破裂导致内容物释放，进一步加重血栓形成。神经损伤诱导蛋白 1（NINJ1）是一种质膜蛋白，通过寡聚化介导膜破裂，这是细胞死亡的典型标志。然而，NINJ1 是否在血小板中表达以及调节血小板功能尚不清楚。本研究旨在评估 NINJ1 在人源和鼠源血小板中的表达，并阐明 NINJ1 在血小板和脓毒症 DIC 中的作用。在这项研究中，使用 NINJ1 阻断肽（NINJ1）来验证 NINJ1 对血小板的体外和体内作用。通过流式细胞术检测血小板 αIIbβ3 和 P-选择素。通过比浊法测量血小板聚集。通过免疫荧光法检测血小板黏附、铺展和 NINJ1 寡聚化。使用盲肠穿孔诱导的脓毒症和 FeCl3 诱导的血栓形成模型来评估 NINJ1 在体内血小板、血栓和 DIC 中的作用。我们发现，抑制 NINJ1 可减轻体外血小板活化。在膜破裂的血小板中验证了 NINJ1 的寡聚化，这是由 PANoptosis 途径调节的。体内研究表明，抑制 NINJ1 可有效减少血小板活化和细胞膜破裂，从而抑制血小板级联反应，导致脓毒症中的抗血栓形成和抗 DIC。这些数据表明，NINJ1 在血小板活化和质膜破裂中起关键作用，抑制 NINJ1 可有效减少血小板依赖性血栓形成和脓毒症中的 DIC。这是首次揭示 NINJ1 在血小板及其相关疾病中的关键作用的研究。
