Department of Hematology, the Third Xiangya Hospital, Central South University, No. 138 Tongzipo Road, Changsha, Hunan, People's Republic of China.
Department of Nutrition, the Third Xiangya Hospital, No. 138 Tongzipo Road, Changsha, Hunan, People's Republic of China.
J Bioenerg Biomembr. 2022 Jun;54(3):155-162. doi: 10.1007/s10863-022-09940-9. Epub 2022 Jun 8.
Thrombocytopenia and impaired platelet function are associated with sepsis-induced organ failure. Numerous studies have shown that mitochondrial ROS (mtROS) and autophagy are related to organ injury in sepsis. However, the relationships between platelet mtROS, autophagy and sepsis organ failure remain unclear. Herein, we explored whether toll like receptor 4 (TLR4) inhibitor alleviates sepsis organ failure by inhibiting platelet mtROS production, autophagy, and GPIIb/IIIa expression.Mice were administrated with LPS, LPS + TAK242 or vehicle. The lungs and kidneys were harvested and analyzed using hematoxylin and eosin staining assay. Platelet rich plasma (PRP) was isolated from blood and platelets aggregation and TLR4 expression were analyzed using flow cytometer and western blot. PRP from healthy volunteers was treated with saline, LPS, or LPS + TAK242, and then mitoSOX and calcium were detected using flow cytometer, and NOX2 and LC3B were tested using western blot.Results showed that TAK242 effectively alleviated LPS-induced acute kidney and lung injury in mice, and decreased CD41 expression more significantly than CD62P. In vitro, by inhibiting TLR4, TAK242 suppressed Ca, mitoSOX fluorescence, NOX2 expression and LC3BII/LC3BI ratio in LPS treated platelets.TLR4 inhibitor TAK242 may effectively alleviate mouse lung and kidney injury by inhibition of mouse platelet GPIIb/IIIa, and reduce LPS-induced mtROS generation related to Ca influx, thus reducing platelet activation.
血小板减少和血小板功能障碍与脓毒症引起的器官衰竭有关。许多研究表明，线粒体 ROS（mtROS）和自噬与脓毒症中的器官损伤有关。然而，血小板 mtROS、自噬与脓毒症器官衰竭之间的关系尚不清楚。在此，我们探讨了 Toll 样受体 4（TLR4）抑制剂是否通过抑制血小板 mtROS 产生、自噬和 GPIIb/IIIa 表达来减轻脓毒症器官衰竭。用 LPS、LPS+TAK242 或载体处理小鼠。用苏木精和伊红染色法分析肺和肾。从血液中分离富含血小板的血浆（PRP），并用流式细胞仪和 Western blot 分析血小板聚集和 TLR4 表达。用生理盐水、LPS 或 LPS+TAK242 处理来自健康志愿者的 PRP，然后用流式细胞仪检测 mitoSOX 和钙，用 Western blot 检测 NOX2 和 LC3B。结果表明，TAK242 有效缓解了 LPS 诱导的小鼠急性肾和肺损伤，且 CD41 表达的降低比 CD62P 更显著。在体外，通过抑制 TLR4，TAK242 抑制了 LPS 处理的血小板中 Ca、mitoSOX 荧光、NOX2 表达和 LC3BII/LC3BI 比值。TLR4 抑制剂 TAK242 通过抑制小鼠血小板 GPIIb/IIIa，可能有效减轻小鼠肺和肾损伤，并减少与 Ca 内流相关的 LPS 诱导的 mtROS 生成，从而减少血小板激活。
