热炎宁合剂在评价人冠状病毒肺炎疫毒袭肺证中的应用

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

前期研究中,该实验室根据国家《新型冠状病毒肺炎诊疗方案》中医临床分型,建立了人冠状病毒肺炎疫毒袭肺证小鼠病证结合模型。该研究采用人冠状病毒肺炎疫毒袭肺证小鼠病证结合模型,评价热炎宁合剂的有效性,为临床应用提供动物实验支持。将小鼠分为正常组,HCoV-229E感染组,寒湿组,寒湿+HCoV-229E感染组,热炎宁大、小剂量组,其中寒湿组、寒湿+HCoV+229E感染组、热炎宁2个剂量组给予寒湿刺激造模7 d,第5天选择HCoV+229E感染组、寒湿+HCoV+229E感染组、热炎宁2个剂量组感染HCoV-229E病毒,感染当天热炎宁给药治疗,给药3 d,第4天取血并解剖取肺组织,计算肺指数和抑制率;流式技术检测外周血液中T和B淋巴细胞百分比;RT-PCR技术检测肺组织核酸病毒载量;ELISA法检测血清中胃动素、胃泌素含量以及肺组织中细胞因子TNF-α,IFN-γ,IL-6,IL-10含量。热炎宁合剂能显著降低人冠状病毒肺炎疫毒袭肺证小鼠的肺指数(P<0.01);能显著提升模型小鼠外周血CD8^+ T淋巴细胞和CD4^+ T淋巴细胞百分比(P<0.05,P<0.01);热炎宁小剂量能有效升高总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),显著降低模型小鼠肺组织中TNF-α,IFN-γ,IL-6,IL-10含量(P<0.01),显著降低模型小鼠血清中胃动素含量(P<0.01)。热炎宁合剂能较好治疗人冠状病毒肺炎疫毒袭肺证,其在改善肺部病变,增强小鼠胃肠道功能,提高小鼠自身免疫机能,降低体内细胞因子表达方面均具有明显疗效,为临床研究提供实验依据。

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.