摘要
目的:利用人血管紧张素转化酶2(hACE2)转基因小鼠模型,进行BD-77抗新型冠状病毒SARS-CoV-2药效学研究,并初步探讨BD-77抗SARS-CoV-2的作用机制。方法:建立SARS-CoV-2奥密克戎(Omicron)和德尔塔(Delta)变异株感染VeroE6细胞模型并给予BD-77,观察BD-77体外抗病毒作用效果;通过试剂盒体外检测BD-77对SARS-CoV-2病毒(Delta/Omicron)刺突S蛋白与血管紧张素转化酶2(ACE2)结合的影响;采用色谱法检测BD-77对新冠病毒S蛋白和N蛋白的结合作;hACE2转基因C57BL/6小鼠分为空白组、SARS-CoV-2感染组、BD-77给药37.5、75 mg·kg^(-1)组,每组各8只,建立SARS-CoV-2感染hACE2转基因小鼠致肺炎模型,观察小鼠生存情况,检测小鼠肺组织病毒滴度并观察肺组织病变情况。结果:BD-77在体外对Omicron和Delta变异株有一定的抑制作用,半数抑制浓度(IC50)分别为526.3、653.0 mg·L^(-1);BD-77对SARS-CoV-2病毒的WT、Omicron和Delta变异株S蛋白与ACE2的结合无明显抑制作用;BD-77与新冠病毒S蛋白、N蛋白无结合作用;空白组小鼠无死亡,SARS-CoV-2感染小鼠死亡率为75%、小鼠肺组织中有大量病毒复制、且肺组织、间质有大面积炎症渗出;与模型组比较,BD-77给药37.5 mg·kg^(-1)组和75 mg·kg^(-1)组能够降低小鼠死亡率、明显降低小鼠肺组织中病毒滴度(P<0.05)并改善肺部病变情况。结论:BD-77显示出在体内外对SARS-CoV-2病毒的显著抑制效果。然而,其作用机制并非直接抑制病毒或干预病毒与宿主的结合过程。这一发现提示了对BD-77的作用机制需要通过进一步的实验来深入研究和阐明。
Objective:The human angiotensin converting enzyme2(hACE2)transgenic mouse model was used to clarify the antiviral efficacy of BD-77 against a novel coronavirus SARS-CoV-2 and explore the action mechanism of BD-77 against SARS-CoV-2.Method:SARS-CoV-2 Omicron and Delta variant strainsinfected VeroE6 cell models were established and administered with BD-77 to observe the antiviral effect of BD-77 in vitro.A kit was used to detect the effect of BD-77 in vitro on the binding of spike S protein of SARS-CoV-2 virus(Delta/Omicron)to angiotensin converting enzyme2(ACE2).Chromatography was adopted to detect the binding of BD-77 to the S protein and N protein of the novel coronavirus.hACE2 transgenic C57BL/6 mice were divided into a blank control group,SARS-CoV-2 infection group,BD-77 administration groups of 37.5 mg·kg^(-1)and 75 mg·kg^(-1),with eight mice in each group.The pneumonia model of SARS-CoV-2-infected hACE2 transgenic mice was built to observe the survival of the mice,detect the virus titer of the lung tissue of the mice,and observe the lesions in the lung tissue.Result:BD-77 had a certain inhibitory effect on Omicron and Delta variant strains in vitro,with median inhibitory concentration(IC50)of 526.3 mg·L^(-1)and 653.0 mg·L^(-1),respectively.BD-77 had no significant inhibitory effect on the binding of the S protein of WT,Omicron,and Delta variant strains of SARS-CoV-2 to ACE2 and had no binding effect with the S protein and N protein of the novel coronavirus.No mice in the blank group died,while the mortality rate of SARS-CoV-2-infected mice was 75%.There was a large amount of virus replication in the lung tissue of the mice and large areas of inflammatory infiltration in the lung tissue and interstitium.Compared with the model group,BD-77 administration groups of 37.5 mg·kg^(-1)and 75 mg·kg^(-1)could reduce the mortality of mice,significantly lower the virus titer in the lung tissue of mice(P<0.05),and improve lung lesions.Conclusion:BD-77 demonstrated significant inhibitory effects against SARS-CoV-2 virus in vitro and in vivo.However,its mechanism of action did not involve direct inhibition of the virus itself or intervention in the virus-host binding process.This finding suggests that the mechanism of action of BD-77 needs to be thoroughly investigated and elucidated by further experiments.
作者
包蕾
马钦海
郭姗姗
赵荣华
夏文
耿子涵
孙静
鲍岩岩
徐洲
严胜龙
肖锦新
陈华荣
黄成钢
崔晓兰
BAO Lei;MA Qinhai;GUO Shanshan;ZHAO Ronghua;XIA Wen;GENG Zihan;SUN Jing;BAO Yanyan;XU Zhou;YAN Shenglong;XIAO Jinxin;CHEN Huarong;HUANG Chenggang;CUI Xiaolan(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Guangzhou Medical University,Guangzhou 511495,China;Medical Sciences Guizhou Bailing Enterprise Group Pharmaceutical Co.Ltd.,Anhun 561099,China;Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai 201203,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第13期45-51,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82141206,82151210)。
