Heme activates platelets and exacerbates rhabdomyolysis-induced acute kidney injury via CLEC-2 and GPVI/FcRγ
Heme activates platelets and exacerbates rhabdomyolysis-induced acute kidney injury via CLEC-2 and GPVI/FcRγ
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DOI：
10.1182/bloodadvances.2020001698
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发表时间：
2021-04-12
期刊：
影响因子：
7.5
通讯作者：
Suzuki-Inoue, Katsue
中科院分区：
文献类型：
--
作者：
Oishi, Saori;Tsukiji, Nagaharu;Suzuki-Inoue, Katsue
越来越多的证据表明，血小板参与除血栓形成和止血以外的多种病理生理过程，如免疫、炎症、胚胎发育和癌症进展。最近的一项研究表明，血红素（氯化血红素）激活的血小板诱导巨噬细胞胞外陷阱（MET），并加剧横纹肌溶解诱导的急性肾损伤（RAKI）;然而，氯化血红素如何激活血小板仍不清楚。在这里，我们报告，C型凝集素样受体-2（CLEC-2）和糖蛋白VI（GPVI）是血小板氯化血红素受体，并参与RAKI的恶化。我们研究了氯化血红素诱导的人和小鼠血小板聚集，氯化血红素与CLEC-2和GPVI的结合，小鼠模型中RAKI相关的表型，以及体外MET的形成。使用蛋白质印迹和表面等离子体共振，我们表明，氯化血红素激活人血小板刺激SYK和PLC γ 2的磷酸化和直接结合CLEC-2和GPVI。此外，氯高铁血红素诱导的鼠血小板聚集在CLEC-2耗尽和FcR γ缺陷（相当于GPVI缺陷）的血小板中部分减少，并且在CLEC-2耗尽的FcR γ缺陷（双敲除）的血小板中几乎完全抑制。此外，氯高铁血红素诱导的鼠血小板聚集被CLEC-2抑制剂钴血卟啉或GPVI抗体（JAQ-1）抑制。肾功能不全，肾小管损伤和MET形成在双敲除RAKI小鼠中减弱。此外，体外MET形成试验表明CLEC-2和GPVI的下游信号通路参与MET形成。我们认为血小板中的CLEC-2和GPVI在RAKI的发展中起重要作用。
There is increasing evidence that platelets participate in multiple pathophysiological processes other than thrombosis and hemostasis, such as immunity, inflammation, embryonic development, and cancer progression. A recent study revealed that heme (hemin)-activated platelets induce macrophage extracellular traps (METs) and exacerbate rhabdomyolysis-induced acute kidney injury (RAKI); however, how hemin activates platelets remains unclear. Here, we report that both C-type lectin-like receptor-2 (CLEC-2) and glycoprotein VI (GPVI) are platelet hemin receptors and are involved in the exacerbation of RAKI. We investigated hemin-induced platelet aggregation in humans and mice, binding of hemin to CLEC-2 and GPVI, the RAKI-associated phenotype in a mouse model, and in vitro MET formation. Using western blotting and surface plasmon resonance, we showed that hemin activates human platelets by stimulating the phosphorylation of SYK and PLC gamma 2 and directly binding to both CLEC-2 and GPVI. Furthermore, hemin-induced murine platelet aggregation was partially reduced in CLEC-2-depleted and FcR gamma-deficient (equivalent to GPVI-deficient) platelets and almost completely inhibited in CLEC-2-depleted FcRy-deficient (double-knockout) platelets. In addition, hemin-induced murine platelet aggregation was inhibited by the CLEC-2 inhibitor cobalt hematoporphyrin or GPVI antibody (JAQ-1). Renal dysfunction, tubular injury, and MET formation were attenuated in double-knockout RAKI mice. Furthermore, in vitro MET formation assay showed that the downstream signaling pathway of CLEC-2 and GPVI is involved in MET formation. We propose that both CLEC-2 and GPVI in platelets play an important role in RAKI development.