Department of Respiratory Medicine, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, China.
Department of Respiratory Medicine, Fangshan Hospital, Beijing University of Chinese Medicine, Beijing 102400, China.
J Tradit Chin Med. 2020 Oct;40(5):803-811. doi: 10.19852/j.cnki.jtcm.2020.05.010.
To investigate synergistic effect of Reduning (RDN) injection plus ribavirin against severe pneumonia induced by H1N1 influenza A virus in mice.
We established a mouse model of severe pneumonia induced by influenza A virus by infecting Balb/c mice with CA07 virus. We randomly assigned the infected mice into four groups, and treated them with normal saline (NS group), RDN (injection, 86.6 mg/kg), ribavirin (injection, 66.6 mg/kg) or double Ribavirin plus RDN group, the same dosage as used in the single treatments) for 5 d. Lung index and lung pathology were recorded or calculated in terms of the curative effective. Cytokines, NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome related protein including caspase-associated recruitment domain (CARD) domain Apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC), caspase-1 and NOD-like receptor family, pyrin domain containing 3 (NLRP3), and reactive oxygen species were simultaneously investigated.
RDN plus ribavirin treatment, not RDN or ribavirin alone, provided a significant survival benefit to the influenza A virus-infected mice. The combination treatment protected the mice against severe influenza infection by attenuating the severe lung injury. The combined treatment also reduced the viral titers in mouse lungs and lung index, downregulated their immunocytokine levels, including IL-1β and IL-18, and down regulated the NLRP3, especially the transcription and translation of caspase-1. Meanwhile NS group had significantly higher reactive oxygen species (ROS) expression which could was dramatically reduced by the treatment of RDN plus ribavirin.
Our study showed that RDN combined with ribavirin could protect the mice, and reduce the lung immunopathologic damage caused by severe influenza pneumonia. The mechanism could be that it reduced ROS produce and inhibited NLRP3 inflammasome activation so that mainly lower the downstream inflammatory cytokines IL-1β and IL-18.
研究热毒宁（RDN）注射液联合利巴韦林对甲型 H1N1 流感病毒诱导的重症肺炎的协同作用。
用 CA07 病毒感染 Balb/c 小鼠建立流感病毒诱导的重症肺炎小鼠模型。将感染小鼠随机分为 4 组，分别用生理盐水（NS 组）、热毒宁（注射液，86.6mg/kg）、利巴韦林（注射液，66.6mg/kg）或利巴韦林加热毒宁（与单药治疗相同剂量）治疗 5d。根据疗效计算肺指数和肺病理。同时检测细胞因子、NOD 样受体家族含pyrin 结构域蛋白 3（NLRP3）炎症小体相关蛋白包括半胱氨酸天冬氨酸蛋白酶募集域（CARD）结构域凋亡相关斑点样蛋白含半胱氨酸蛋白酶募集域（ASC）、半胱氨酸天冬氨酸蛋白酶-1 和 NOD 样受体家族，pyrin 结构域包含 3（NLRP3）和活性氧（ROS）。
热毒宁联合利巴韦林治疗，而非热毒宁或利巴韦林单独治疗，可显著提高甲型流感病毒感染小鼠的生存率。联合治疗通过减轻严重的肺损伤来保护小鼠免受严重流感感染。联合治疗还降低了小鼠肺部的病毒滴度和肺指数，下调了免疫细胞因子水平，包括白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18），并下调了 NLRP3，特别是半胱氨酸天冬氨酸蛋白酶-1 的转录和翻译。同时，NS 组的活性氧（ROS）表达明显升高，热毒宁联合利巴韦林治疗可显著降低其表达。
本研究表明，热毒宁联合利巴韦林可保护小鼠，减轻严重流感肺炎引起的肺免疫病理损伤。其机制可能是降低 ROS 产生，抑制 NLRP3 炎症小体激活，从而降低下游炎症细胞因子 IL-1β 和 IL-18。
