Liu Anding, Weiss Stefanie, Fang Haoshu, Claus Ralf A, Rödel Jürgen, Dirsch Olaf, Dahmen Uta
*Experimental Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; †Experimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, Jena, Germany; ‡Department of Pathophysiology, Anhui Medical University, Hefei, China; §Integrated Research and Treatment Center, Center for Sepsis Control and Care (CSCC), Jena University Hospital, Jena, Germany; ∥Institute of Medical Microbiology, Jena University Hospital, Jena, Germany; and ¶Institute for Pathology, Jena University Hospital, Jena, Germany.
Shock. 2015 May;43(5):497-503. doi: 10.1097/SHK.0000000000000338.
The effect of granulocyte colony-stimulating factor (G-CSF) on sepsis is discussed controversially in clinical studies. We previously demonstrated that G-CSF treatment induced lipopolysaccharide (LPS) sensitization via up-regulation of LPS-binding protein (LBP). We hypothesized that the futile effect of G-CSF-treatment in sepsis might be due to its ability to up-regulate LBP. Therefore, blockade of LBP may attenuate the G-CSF-induced LPS sensitization and protect animals from polymicrobial sepsis. Endogenous LBP levels were up-regulated by pretreatment with G-CSF, and the LBP protein was blocked by administration of a specific blocking peptide-LBPK95A. Polymicrobial sepsis was induced by intraperitoneal injection of feces slurry. Rats were monitored every 3 up to 72 h to observe the survival rate. Tissue injury, bacterial infiltration, local inflammatory response, and neutrophil infiltration at 0, 2, and 12 h after the septic insult were analyzed. The survival benefit of G-CSF pretreatment was improved when combined with LBPK95A treatment (control vs. G-CSF vs. combi: 36% vs. 56% vs. 93%; P < 0.05). Combined treatment of G-CSF and LBPK95A was associated with the minimal tissue damage. Treatment with LBPK95A significantly inhibited the neutrophil infiltration without interfering with the bacterial clearance. The G-CSF-induced inflammatory sensitization effect was inhibited by LBPK95A, indicated by the decrease of cytokines expression, and the activation of nuclear factor kappa B and signal transducer and activator of transcription 3 signaling pathway. In conclusion, these results suggested that the effect of prophylactic augmentation of the host's response via G-CSF pretreatment was further enhanced by inhibition of the up-regulation of LBP.
粒细胞集落刺激因子（G-CSF）对脓毒症的影响在临床研究中存在争议。我们之前证明，G-CSF治疗通过上调脂多糖（LPS）结合蛋白（LBP）诱导LPS致敏。我们推测，G-CSF治疗在脓毒症中产生无效作用可能是由于其上调LBP的能力。因此，阻断LBP可能减弱G-CSF诱导的LPS致敏，并保护动物免受多微生物脓毒症的影响。通过G-CSF预处理上调内源性LBP水平，并通过给予特异性阻断肽-LBPK95A来阻断LBP蛋白。通过腹腔注射粪便悬液诱导多微生物脓毒症。每3小时监测大鼠直至72小时，以观察存活率。分析脓毒症攻击后0、2和12小时的组织损伤、细菌浸润、局部炎症反应和中性粒细胞浸润。当与LBPK95A治疗联合使用时，G-CSF预处理的生存益处得到改善（对照组 vs. G-CSF组 vs. 联合治疗组：36% vs. 56% vs. 93%；P < 0.05）。G-CSF和LBPK95A的联合治疗与最小的组织损伤相关。LBPK95A治疗显著抑制中性粒细胞浸润，而不干扰细菌清除。细胞因子表达的降低以及核因子κB和信号转导子及转录激活子3信号通路的激活表明，LBPK95A抑制了G-CSF诱导的炎症致敏作用。总之，这些结果表明，通过抑制LBP的上调，可进一步增强通过G-CSF预处理预防性增强宿主反应的效果。
