Liu Zhaohui, Wang Zhengping, Han Guangwei, Huang Lina, Jiang Jihong, Li Shitong
Department of Anesthesiology, Cangzhou Central Hospital, Hebei Medical University, Hebei, China.
Department of Anesthesiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China.
J Surg Res. 2016 Feb;200(2):593-603. doi: 10.1016/j.jss.2015.08.032. Epub 2015 Aug 28.
High mobility group box-1 (HMGB1) acts as an inflammatory mediator and has been implicated in pathophysiological damage of vascular inflammatory diseases. Ketamine, an anesthetic agent with sedative and analgesic properties, has been shown to have potent anti-inflammatory effects in a variety of models of systemic inflammation. However, the effects of ketamine on HMGB1-mediated proinflammatory responses have not been fully investigated. In the present study, we investigated the effects of ketamine on HMGB1-activated endothelial cells and explored the underlying mechanisms.
Human endothelial cells were incubated with or without HMGB1 (1 μg/mL) in the presence or absence of ketamine, an nuclear factor (NF)-κB inhibitor (PDTC), anti-toll-like receptor (TLR)2/4 antibody, or small interfering RNA (siRNA). The anti-inflammatory activities of ketamine were determined by measuring solute flux, leukocyte adhesion and migration, and activation of proinflammatory proteins in HMGB1-activated endothelial cells. The effect of ketamine on TLR-2/4 and NF-κB activation was evaluated using enzyme-linked immunosorbent assays and immunofluorescence confocal microscopy assay.
We found that ketamine inhibited the HMGB1-mediated barrier disruption, neutrophil adhesion and migration, and expression of cell adhesion molecules in a dose-dependent manner. Furthermore, ketamine downregulated the TLR-2 and -4, expression in HMGB1-activated endothelial cells. Treatment with ketamine also significantly inhibited the activation of TLR2/4 and the nuclear translocation of NF-κB p50/p65. Furthermore, our study shows that the HMGB1-induced release of inflammatory mediators was suppressed by PDTC, anti-TLR2/4 antibody, and siRNA.
Our study has demonstrated that ketamine exerts anti-inflammatory effects in HMGB1-mediated proinflammatory responses in a dose-dependent manner. The mechanism responsible for these effects involves the TLR2/4 and NF-κB signaling pathway.
高迁移率族蛋白B1（HMGB1）作为一种炎症介质，与血管炎性疾病的病理生理损伤有关。氯胺酮是一种具有镇静和镇痛特性的麻醉剂，已被证明在多种全身炎症模型中具有强大的抗炎作用。然而，氯胺酮对HMGB1介导的促炎反应的影响尚未得到充分研究。在本研究中，我们研究了氯胺酮对HMGB1激活的内皮细胞的影响，并探讨了其潜在机制。
在有或无氯胺酮、核因子（NF）-κB抑制剂（PDTC）、抗Toll样受体（TLR）2/4抗体或小干扰RNA（siRNA）的情况下，将人内皮细胞与HMGB1（1μg/mL）一起或不与HMGB1一起孵育。通过测量溶质通量、白细胞粘附和迁移以及HMGB1激活的内皮细胞中促炎蛋白的激活来确定氯胺酮的抗炎活性。使用酶联免疫吸附测定和免疫荧光共聚焦显微镜测定法评估氯胺酮对TLR-2/4和NF-κB激活的影响。
我们发现氯胺酮以剂量依赖性方式抑制HMGB1介导的屏障破坏、中性粒细胞粘附和迁移以及细胞粘附分子的表达。此外，氯胺酮下调了HMGB1激活的内皮细胞中TLR-2和-4的表达。氯胺酮治疗还显著抑制了TLR2/4的激活和NF-κB p50/p65的核转位。此外，我们的研究表明，PDTC、抗TLR2/4抗体和siRNA抑制了HMGB1诱导的炎症介质释放。
我们的研究表明，氯胺酮在HMGB1介导的促炎反应中以剂量依赖性方式发挥抗炎作用。这些作用的机制涉及TLR2/4和NF-κB信号通路。
