School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Toxicol Appl Pharmacol. 2020 Apr 1;392:114921. doi: 10.1016/j.taap.2020.114921. Epub 2020 Feb 12.
Angioedema may occur during local anesthetic (LA) injection in the perioperative period. Histaminergic angioedema is the most common form of angioedema. It has been reported that LA is a potential exogenous ligand for histamine receptor 1 (HR). Whether HR participates in LA-induced angioedema is still controversial. By using a constructed HR high-expressed cell model, siRNA transfection, pharmacologic means, and genetically modified animal models, here we showed that HR mediated LA-induced hyperpermeability. LA with uncycled N-methyl scaffold in the side chain (procaine, tetracaine and lidocaine) had a better strength of drug-HR affinity than that for LA with cycled N atom (bupivacaine and ropivacaine) by the molecular docking assay and equilibrium dissociation constant (K values) obtained from the cell membrane chromatography (CMC) relative standard method. Procaine, tetracaine, and lidocaine triggered big calcium mobilization in HR-HEK293 cells and human umbilical vein endothelial cells (HUVECs) but much weaker in NC-HEK293 cells or HR knockdown HUVECs. Besides, the results of transendothelial resistance measurement, paracellular flux assay and immunofluorescence showed that procaine induced HR-dependent hyperpermeability, which involved in PLCγ/IP3R/PKC, ERK1/2, Akt signaling pathways, downstream vascular endothelial cadherin (VE-cad) destabilization. Furthermore, HR gene knockout prevented paw swelling and vascular leakage caused by procaine, tetracaine, and lidocaine in vivo. This study supported a key role of HR in LA-induced angioedema, and suggested that in the design of LA structure, the ring formation of the N-methyl scaffold on the side chain can properly avoid the angioedema.
血管性水肿可发生在围手术期局部麻醉（LA）注射期间。组织胺能血管性水肿是最常见的血管性水肿形式。据报道，LA 是组胺受体 1（HR）的潜在外源性配体。HR 是否参与 LA 诱导的血管性水肿仍存在争议。通过构建 HR 高表达细胞模型、siRNA 转染、药理手段和基因修饰动物模型，我们发现 HR 介导了 LA 诱导的高通透性。通过分子对接和细胞膜色谱（CMC）相对标准法获得的平衡解离常数（K 值），侧链中无环 N-甲基支架的 LA（普鲁卡因、丁卡因和利多卡因）与具有环 N 原子的 LA（布比卡因和罗哌卡因）相比，具有更强的药物-HR 亲和力。普鲁卡因、丁卡因和利多卡因在 HR-HEK293 细胞和人脐静脉内皮细胞（HUVEC）中引发大量钙动员，但在 NC-HEK293 细胞或 HR 敲低 HUVEC 中则弱得多。此外，跨内皮电阻测量、细胞旁通量测定和免疫荧光结果表明，普鲁卡因诱导的 HR 依赖性高通透性涉及 PLCγ/IP3R/PKC、ERK1/2、Akt 信号通路，下游血管内皮钙黏蛋白（VE-cad）不稳定。此外，HR 基因敲除可防止普鲁卡因、丁卡因和利多卡因体内引起的爪肿胀和血管渗漏。这项研究支持 HR 在 LA 诱导的血管性水肿中的关键作用，并表明在 LA 结构设计中，侧链上 N-甲基支架的环形成可以适当避免血管性水肿。
