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热炎宁合剂在评估新型冠状病毒肺炎合并疫病袭肺证中的应用

[Application of Reyanning Mixture in evaluating combining disease with syndrome of human coronavirus pneumonia with pestilence attacking lung syndrome].

作者信息

Bao Lei, Shi Yu-Jing, Geng Zi-Han, Sun Jing, Zhao Rong-Hua, DU Cheng-Qiang, Chu Ya-Jun, Cui Xiao-Lan

机构信息

Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.

Tsing Hua De Ren Xi'an Happiness Pharmaceutical Co.,Ltd. Xi'an 710043,China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2020 Apr;45(7):1465-1472. doi: 10.19540/j.cnki.cjcmm.20200303.401.

Abstract

In the previous research, our laboratory established a mouse model combining disease with syndrome of human coronavi-rus pneumonia with pestilence attacking the lung syndrome, based on the national traditional Chinese medicine clinical classification of Novel Coronavirus Infected Pneumonia Diagnosis and Treatment Plan. In this study, a mouse model combining disease with syndrome of human coronavirus pneumonia with pestilence attacking the lung syndrome was used to evaluate the effectiveness of Reyanning Mixture to provide animal experimental support for clinical application. Mice were divided into normal group, 229 E infection group, cold-dampness group, cold-dampness+229 E infection group(the model group), Reyanning high and low dose groups. The cold-dampness group, cold-dampness+229 E infection group, two Reyanning groups were given cold and damp stimulation for 7 days. On the 5 th day, the 229 E infection group, cold-dampness+229 E infection group, and two Reyanning groups were infected with HCoV-229 E virus. Reyanning was administered for 3 days, starting from the day of infection. Blood was collected on the 4 th day and the lung tissue was dissected to calculate the lung index and inhibition rate; flow cytometry was used to detect the percentage of T and B lymphocytes in peripheral blood; RT-PCR was used to detect the nucleic acid virus load in lung tissue; ELISA was used to detect motilin and gastrin in serum, and inflammatory factors TNF-α, IFN-γ, IL-6, IL-10 in lung tissue proteins. Reyanning Mixture could reduce the lung index(P<0.01) of coronavirus pneumonia mice with pestilence attacking the lung; it could significantly increase the percentage of CD8+ T lymphocytes and CD4+ T lymphocytes in peripheral blood of model mice(P<0.05, P<0.01). The low dose of Reyanning could effectively increase the percentage of total B lymphocytes(P<0.05), reduce virus load in lung tissue of model mice(P<0.01), reduce the levels of TNF-α, IFN-γ, IL-6, IL-10 in the lung tissue of model mice(P<0.01), reduce the content of motilin in the serum of model mice(P<0.01). Reyanning Mixture convey a better effect in treating coronavirus pneumonia mice with pestilence attacking the lung. It manifested obvious effects in improving lung lesions, enhancing the gastrointestinal function of mice, improving the autoimmune function of mice, and reducing the expression of inflammatory factors in vivo, which could provide evidences for clinical research.

摘要

在前期研究中,本实验室依据国家中医药《新型冠状病毒感染的肺炎诊疗方案》临床分类,建立了人冠状病毒肺炎病与疫毒犯肺证相结合的小鼠模型。本研究运用人冠状病毒肺炎病与疫毒犯肺证相结合的小鼠模型,评价热炎宁合剂的有效性,为临床应用提供动物实验依据。将小鼠分为正常组、229E感染组、寒湿组、寒湿+229E感染组(模型组)、热炎宁高、低剂量组。对寒湿组、寒湿+229E感染组、热炎宁两个组进行7天的寒湿刺激。第5天,对229E感染组、寒湿+229E感染组、热炎宁两个组感染HCoV-229E病毒。从感染当天开始给予热炎宁,连续给药3天。第4天采血并解剖肺组织,计算肺指数和抑制率;采用流式细胞术检测外周血T、B淋巴细胞百分比;采用RT-PCR检测肺组织核酸病毒载量;采用ELISA检测血清胃动素、胃泌素及肺组织蛋白中炎性因子TNF-α、IFN-γ、IL-6、IL-10。热炎宁合剂可降低疫毒犯肺型冠状病毒肺炎小鼠的肺指数(P<0.01);可显著提高模型小鼠外周血CD8⁺T淋巴细胞及CD4⁺T淋巴细胞百分比(P<0.05,P<0.01)。热炎宁低剂量可有效提高总B淋巴细胞百分比(P<0.05),降低模型小鼠肺组织病毒载量(P<0.01),降低模型小鼠肺组织TNF-α、IFN-γ、IL-6、IL-10水平(P<0.01),降低模型小鼠血清胃动素含量(P<0.01)。热炎宁合剂对疫毒犯肺型冠状病毒肺炎小鼠有较好的治疗作用。在改善肺部病变、增强小鼠胃肠功能、提高小鼠自身免疫功能、降低体内炎性因子表达等方面表现出明显效果,可为临床研究提供依据。

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