Wang Hao-Min, Huang Ping, Li Quan, Yan Lu-Lu, Sun Kai, Yan Li, Pan Chun-Shui, Wei Xiao-Hong, Liu Yu-Ying, Hu Bai-He, Wang Chuan-She, Fan Jing-Yu, Han Jing-Yan
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China.
Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China.
Front Physiol. 2019 Oct 25;10:1320. doi: 10.3389/fphys.2019.01320. eCollection 2019.
: Lipopolysaccharide (LPS) causes microvascular dysfunction, which is a key episode in the pathogenesis of endotoxemia. This work aimed to investigate the effect of Qing-Ying-Tang (QYT), a compound Chinese medicine in cerebral microcirculation disturbance and brain damage induced by LPS. : Male C57/BL6 mice were continuously transfused with LPS (7.5 mg/kg/h) through the left femoral vein for 2 h. QYT (14.3 g/kg) was given orally 2 h after LPS administration. The dynamics of cerebral microcirculation were evaluated by intravital microscopy. Brain tissue edema was assessed by brain water content and Evans Blue leakage. Cytokines in plasma and brain were evaluated by flow cytometry. Confocal microscopy and Western blot were applied to detect the expression of junction and adhesion proteins, and signaling proteins concerned in mouse brain tissue. : Post-treatment with QYT significantly ameliorated LPS-induced leukocyte adhesion to microvascular wall and albumin leakage from cerebral venules and brain tissue edema, attenuated the increase of MCP-1, MIP-1α, IL-1α, IL-6, and VCAM-1 in brain tissue and the activation of NF-κB and expression of MMP-9 in brain. QYT ameliorated the downregulation of claudin-5, occludin, JAM-1, ZO-1, collagen IV as well as the expression and phosphorylation of VE-cadherin in mouse brain. : This study demonstrated that QYT protected cerebral microvascular barrier from disruption after LPS by acting on the transcellular pathway mediated by caveolae and paracellular pathway mediated by junction proteins. This result suggests QYT as a potential strategy to deal with endotoxemia.
脂多糖(LPS)可导致微血管功能障碍,这是内毒素血症发病机制中的关键环节。本研究旨在探讨中药复方清营汤(QYT)对LPS诱导的脑微循环障碍和脑损伤的影响。将雄性C57/BL6小鼠通过左股静脉连续输注LPS(7.5mg/kg/h)2小时。在给予LPS 2小时后口服QYT(14.3g/kg)。通过活体显微镜评估脑微循环的动态变化。通过脑含水量和伊文思蓝渗漏评估脑组织水肿。通过流式细胞术评估血浆和脑中的细胞因子。应用共聚焦显微镜和蛋白质免疫印迹法检测小鼠脑组织中连接蛋白、黏附蛋白及相关信号蛋白的表达。QYT治疗后可显著改善LPS诱导的白细胞黏附于微血管壁、脑静脉白蛋白渗漏和脑组织水肿,减轻脑组织中MCP-1、MIP-1α、IL-1α、IL-6和VCAM-1的增加以及脑中NF-κB的激活和MMP-9的表达。QYT改善了小鼠脑中claudin-5、occludin、JAM-1、ZO-1、IV型胶原的下调以及VE-钙黏蛋白的表达和磷酸化。本研究表明QYT通过作用于小窝介导 的跨细胞途径和连接蛋白介导的细胞旁途径保护LPS作用后的脑微血管屏障。该结果提示QYT可能是一种应对内毒素血症的潜在策略。