Wang Aili, Zheng Ni, Jia Qing, Chen Yu, Xu Shun
Department of Burn and Plastic Surgery, The Seventh People's Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200137, P.R. China.
Department of Nuclear Medicine, The Seventh People's Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200137, P.R. China.
Exp Ther Med. 2022 Sep 1;24(5):646. doi: 10.3892/etm.2022.11583. eCollection 2022 Nov.
S100 calcium-binding protein A9 (S100A9) is highly expressed in the serum exosomes of patients with burn injuries. The present study aimed to investigate the underlying mechanisms of burn injury-associated exosomes in the regulation of myocardial cell pyroptosis. Reverse transcription-quantitative PCR and western blotting were used to examine relative mRNA and protein levels. The morphology of exosomes was visualized using transmission electron microscopy. The expression levels of IL-1β and IL-18 in cells were examined using ELISA kits. Finally, cell pyroptosis was examined using flow cytometry. When AC16 cells were treated with the serum exosomes obtained from patients with burn injuries, pyroptosis was significantly promoted and the expression levels of IL-1β and IL-18 were increased. NLR family pyrin domain containing 3 (NLRP3), S100A9, caspase-1 and Gasdermin D (GSDMD)-N expression levels were also upregulated. However, these were significantly reversed by anti-S100A9 antibodies. Thereafter, CY-09, an NLRP3 inhibitor, was revealed to restore the increase in pyroptosis and IL-18, IL-1β, caspase-1, NLRP3 and GSDMD-N expression levels caused by recombinant S100A9 to be similar to the control. These findings suggested that burn injury-associated exosomes containing S100A9 can affect AC16 cell pyroptosis through NLRP3.
S100钙结合蛋白A9（S100A9）在烧伤患者的血清外泌体中高表达。本研究旨在探讨烧伤相关外泌体调控心肌细胞焦亡的潜在机制。采用逆转录定量PCR和蛋白质印迹法检测相关mRNA和蛋白质水平。利用透射电子显微镜观察外泌体的形态。使用酶联免疫吸附测定试剂盒检测细胞中白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）的表达水平。最后，采用流式细胞术检测细胞焦亡情况。当用烧伤患者的血清外泌体处理AC16细胞时，焦亡显著增加，IL-1β和IL-18的表达水平升高。含NLR家族pyrin结构域蛋白3（NLRP3）、S100A9、半胱天冬酶-1（caspase-1）和Gasdermin D（GSDMD）-N的表达水平也上调。然而，抗S100A9抗体可显著逆转这些变化。此后，研究发现NLRP3抑制剂CY-09可使重组S100A9引起的焦亡增加以及IL-18、IL-1β、caspase-1、NLRP3和GSDMD-N表达水平恢复至与对照组相似的水平。这些研究结果表明，含有S100A9的烧伤相关外泌体可通过NLRP3影响AC16细胞焦亡。
