Department of Critical Care Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Peking Union Medical College, 1 Shuaifuyuan, Dongcheng District, Beijing, 100730, China.
Inflammation. 2018 Dec;41(6):2030-2040. doi: 10.1007/s10753-018-0846-z.
This study aimed to investigate the effect of probenecid (Pro) as an inhibitor of the pannexin-1 (Panx-1) channel-mediated release of intracellular ATP to the extracellular compartment on inflammation, cellular energy crisis, and organ injury in a rabbit sepsis model induced by Escherichia coli lipopolysaccharides (LPS). A total of 24 anesthetized and ventilated rabbits were randomly assigned to receive one of four treatments: infusion of LPS without Pro (LPS group), infusion of LPS with Pro (LPS + Pro group), sham operation without Pro (normal group), and sham operation with Pro (normal + Pro group). The LPS group had significantly higher serum ATP levels, serum inflammatory factor levels (TNF-α, IL-6, and IL-1β), and lower ATP concentrations and ATP/ADP ratios in the skeletal muscle tissue than the normal group. Compared to that at baseline, the expression of Panx-1 in peripheral blood cells increased significantly after the infusion of LPS (fluorescence intensity of Panx-1: T0 (baseline) vs. T1 (post-LPS) = 10 ± 1.2 vs. 84 ± 48, P < 0.0001; paired differences 73 ± 46, P = 0.024). Moreover, the LPS group exhibited higher expression of Panx-1 in the skeletal muscle tissue than the normal group. The serum ATP level was significantly positively correlated with IL-1β (R = 0.602, P = 0.001), IL-6 (R = 0.381, P = 0.033), and TNF-α (R = 0.514, P = 0.005) in 24 paired measurements. Compared to the LPS group, the LPS + Pro group had significantly lower levels of inflammatory factors (TNF-α, IL-6, and IL-1β) and serum ATP. In the skeletal muscle tissue, the LPS + Pro group also had a higher ATP concentration (1.1 ± 0.15 vs. 1.33 ± 0.17, P = 0.041) and ATP/ADP ratio (0.37 ± 0.03 vs. 0.51 ± 0.06, P = 0.002) and a lower histopathological damage score (4.67 ± 0.52 vs. 3 ± 0.63, P = 0.004). An overexpression of Panx-1 channel might be responsible for the strong inflammatory response, high serum ATP level, and skeletal muscle cellular energy crisis and histopathological damages in sepsis. Inhibiting Panx-1 channel-mediated release of intracellular ATP could decrease the above-mentioned injuries, and Panx-1 might be a potential therapeutic target in sepsis.
本研究旨在探讨丙磺舒（Probenecid，Pro）作为缝隙连接蛋白 1（Panx-1）通道介导的细胞内 ATP 向细胞外间隙释放的抑制剂，对大肠埃希菌脂多糖（LPS）诱导的兔脓毒症模型中的炎症、细胞能量危机和器官损伤的影响。共有 24 只麻醉和通气的兔子被随机分为四组：不给予 Pro 的 LPS 输注（LPS 组）、给予 Pro 的 LPS 输注（LPS + Pro 组）、不给予 Pro 的假手术（正常组）和给予 Pro 的假手术（正常 + Pro 组）。与正常组相比，LPS 组的血清 ATP 水平、血清炎症因子水平（TNF-α、IL-6 和 IL-1β）显著升高，骨骼肌组织中的 ATP 浓度和 ATP/ADP 比值显著降低。与基础值相比，LPS 输注后外周血中 Panx-1 的表达显著增加（Panx-1 的荧光强度：T0（基线）vs. T1（LPS 后）= 10 ± 1.2 vs. 84 ± 48，P < 0.0001；配对差异 73 ± 46，P = 0.024）。此外，LPS 组的骨骼肌组织中 Panx-1 的表达也高于正常组。血清 ATP 水平与 IL-1β（R = 0.602，P = 0.001）、IL-6（R = 0.381，P = 0.033）和 TNF-α（R = 0.514，P = 0.005）在 24 对测量中呈显著正相关。与 LPS 组相比，LPS + Pro 组的炎症因子（TNF-α、IL-6 和 IL-1β）和血清 ATP 水平显著降低。在骨骼肌组织中，LPS + Pro 组的 ATP 浓度（1.1 ± 0.15 vs. 1.33 ± 0.17，P = 0.041）和 ATP/ADP 比值（0.37 ± 0.03 vs. 0.51 ± 0.06，P = 0.002）更高，组织病理学损伤评分（4.67 ± 0.52 vs. 3 ± 0.63，P = 0.004）更低。缝隙连接蛋白 1 通道的过度表达可能是脓毒症中强烈的炎症反应、高血清 ATP 水平和骨骼肌细胞能量危机和组织病理学损伤的原因。抑制缝隙连接蛋白 1 通道介导的细胞内 ATP 释放可以减少上述损伤，缝隙连接蛋白 1 可能是脓毒症的一个潜在治疗靶点。
