苦参碱氯化钠注射液对人冠状病毒肺炎寒湿疫毒袭肺证小鼠病证结合模型的治疗作用

Effect of matrine sodium chloride injection on a mouse model combining disease with syndrome of human coronavirus pneumonia with cold-dampness pestilence attacking the lung

"疫毒袭肺证"是北京市中医管理局对新型冠状病毒感染肺炎的中医病证分类,本研究首次建立了与新型冠状病毒中医寒湿疫毒袭肺证相吻合的病证结合小鼠模型,并基于免疫调节及炎性损伤等环节,研究了疫区临床用药苦参碱氯化钠注射液对人冠状病毒肺炎寒湿疫毒袭肺证小鼠病证结合模型的治疗作用。通过肺指数、肺指数抑制率和肺组织病理检测评价苦参碱氯化钠注射液对模型小鼠的治疗作用;RT-PCR检测肺组织病毒核酸表达量评价其抗病毒作用;流式细胞术检测CD4+T细胞、CD8+T细胞及B细胞的百分比评价其免疫调节作用;ELISA法检测白细胞介素-6(IL-6)、IL-10、肿瘤坏死因子-α(TNF-α)及γ干扰素(IFN-γ)的表达量评价其抗炎作用。实验中所有操作均遵循美国国立卫生研究院(NIH)及北京市实验动物伦理委员会的规定,并经过中国中医科学院中药研究所动物伦理委员会批准。结果显示,苦参碱氯化钠注射液高剂量组(36.67 mL·kg-1·d-1)和低剂量组(18.33 mL·kg-1·d-1)腹腔注射给药显著减轻模型小鼠肺组织病理损伤,肺指数抑制率分别为86.86%和76.53%;降低肺组织中IL-6、IL-10、TNF-α和IFN-γ的含量及肺组织中病毒载量;升高外周血CD4+T细胞、CD8+T细胞及B细胞的百分比。以上结果表明,苦参碱氯化钠注射液对人冠状病毒肺炎寒湿疫毒袭肺证小鼠病证结合模型具有显著的治疗作用,其机制与抑制病毒复制、调节免疫功能、抑制炎症因子释放等作用有关。本研究为苦参碱氯化钠注射液临床用药提供了实验室数据支持,研究结果显示该药在开发为防治新型冠状病毒肺炎药物方面具有很好的应用前景,对现阶段抗击新型冠状病毒肺炎具有重要意义。

"TCM syndrome of plague attack lung"is a classification of traditional Chinese medicine syndromes of the novel coronavirus pneumonia by the Beijing Municipal Administration of Traditional Chinese Medicine.In this study,a mouse model combining disease with syndrome of human coronavirus pneumonia with cold-dampness pestilence attacking the lung was established for the first time,and the therapeutic effect of matrine sodium chloride injection was evaluated based on immune regulation and inflammatory damage.Lung index,lung index inhibition rate and HE stain were used to evaluate the therapeutic effect of matrine sodium chloride injection on the model mice;the viral load in lung tissue was measured by RT-PCR to evaluate its antiviral effect;the percentage of CD4+T cells,CD8+T cells and B cells were detected by flow cytometry to evaluate its immunomodulatory effect;the production of interleukin 6(IL-6),IL-10,tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)were measured by ELISA to evaluate its anti-inflammatory effect.All interventions and operations in the experiment were approved by the Animal Ethics Committee of the Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,and conformed to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health(NIH)and Beijing Experimental Animal Ethics Committee.The results showed that intraperitoneal injection of the high-dose(36.67 mL·kg-1·d-1)and low-dose(18.33 mL·kg-1·d-1)of matrine sodium chloride injection significantly improved the pathological damage of lung tissue and reduced lung index.The lung index inhibition rates were 86.86%and 76.53%,respectively.The production of IL-6,IL-10,TNF-α,IFN-γ,as well as the viral load in lung tissue were reduced significantly compared to the model;the percentage of CD4+T cells,CD8+T cells and B cells in peripheral blood were increased compared to the model.These results indicated that the matrine sodium chloride injection has an evident therapeutic effect on the model,and its mechanism was related to the inhibition virus replication,regulation of immunity function and inhibition of inflammatory factor release.This study provided laboratory data support for matrine sodium chloride injection which was used to treat the novel coronavirus pneumonia in clinical in Hubei province.These results indicated that the matrine sodium chloride injection has a good prospect for prevention and treatment of the novel coronavirus pneumonia.