Han Jiayin, Zhao Yong, Zhang Yushi, Li Chunying, Yi Yan, Pan Chen, Tian Jingzhuo, Yang Yifei, Cui Hongyu, Wang Lianmei, Liu Suyan, Liu Jing, Deng Nuo, Liang Aihua
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China.
Front Pharmacol. 2018 Feb 12;9:87. doi: 10.3389/fphar.2018.00087. eCollection 2018.
Shuanghuanglian injection (SHLI) is a famous Chinese medicine used as an intravenous preparation for the treatment of acute respiratory tract infections. In the recent years, the immediate hypersensitivity reactions induced by SHLI have attracted broad attention. However, the mechanism involved in these reactions has not yet been elucidated. The present study aims to explore the characteristics of the immediate hypersensitivity reactions induced by SHLI and deciphers the role of the RhoA/ROCK signaling pathway in these reactions. SHLI-immunized mice or naive mice were intravenously injected (i.v.) with SHLI (600 mg/kg) once, and vascular leakage in the ears was evaluated. Passive cutaneous anaphylaxis test was conducted using sera collected from SHLI-immunized mice. Naive mice were administered (i.v.) with a single dose of 150, 300, or 600 mg/kg of SHLI, and vascular leakage, histamine release, and histopathological alterations in the ears, lungs, and intestines were tested. , human umbilical vein endothelial cell (HUVEC) monolayer was incubated with SHLI (0.05, 0.1, or 0.15 mg/mL), and the changes in endothelial permeability and cytoskeleton were observed. Western blot analysis was performed and ROCK inhibitor was employed to investigate the contribution of the RhoA/ROCK signaling pathway in SHLI-induced hypersensitivity reactions, both in HUVECs and in mice. Our results indicate that SHLI was able to cause immediate dose-dependent vascular leakage, edema, and exudates in the ears, lungs, and intestines, and histamine release in mice. These were pseudo-allergic reactions, as SHLI-specific IgE was not elicited during sensitization. In addition, SHLI induced reorganization of actin cytoskeleton and disrupted the endothelial barrier. The administration of SHLI directly activated the RhoA/ROCK signaling pathway both in HUVECs and in the ears, lungs, and intestines of mice. Fasudil hydrochloride, a ROCK inhibitor, ameliorated the SHLI-induced hypersensitivity reactions in both endothelial cells and mice indicating its protective effect. SHLI-induced pseudo-allergic reactions were mediated by the activation of the RhoA/ROCK signaling pathway. : This study presents a novel mechanism of SHLI-induced immediate hypersensitivity reactions and suggests a potential therapeutic strategy to prevent the associated adverse reactions.
双黄连注射液（SHLI）是一种著名的中药，用作静脉制剂治疗急性呼吸道感染。近年来，SHLI引起的速发型超敏反应引起了广泛关注。然而，这些反应所涉及的机制尚未阐明。本研究旨在探讨SHLI诱导的速发型超敏反应的特征，并解读RhoA/ROCK信号通路在这些反应中的作用。将经SHLI免疫的小鼠或未免疫的小鼠静脉注射（i.v.）一次SHLI（600mg/kg），并评估耳部的血管渗漏。使用从经SHLI免疫的小鼠收集的血清进行被动皮肤过敏试验。给未免疫的小鼠静脉注射单剂量150、300或600mg/kg的SHLI，并检测耳部、肺部和肠道的血管渗漏、组胺释放和组织病理学改变。将人脐静脉内皮细胞（HUVEC）单层与SHLI（0.05、0.1或0.15mg/mL）孵育，观察内皮通透性和细胞骨架的变化。进行蛋白质印迹分析并使用ROCK抑制剂研究RhoA/ROCK信号通路在HUVEC和小鼠中SHLI诱导的超敏反应中的作用。我们的结果表明，SHLI能够在小鼠的耳部、肺部和肠道中引起即刻的剂量依赖性血管渗漏、水肿和渗出物以及组胺释放。这些是假过敏反应，因为在致敏过程中未引发SHLI特异性IgE。此外，SHLI诱导肌动蛋白细胞骨架重组并破坏内皮屏障。SHLI的给药在HUVEC以及小鼠的耳部、肺部和肠道中直接激活了RhoA/ROCK信号通路。盐酸法舒地尔，一种ROCK抑制剂，改善了SHLI在内皮细胞和小鼠中诱导的超敏反应，表明其保护作用。SHLI诱导的假过敏反应是由RhoA/ROCK信号通路的激活介导的。：本研究提出了SHLI诱导的速发型超敏反应的新机制，并提出了预防相关不良反应的潜在治疗策略。
