中药有效成分对人肺泡Ⅱ型肺泡细胞ACE2作用的研究  

Study on the effects of active ingredients of traditional Chinese medicine on ACE2 of human alveolar typeⅡcells

作　　者：庞晓军[1,2] 黎东旺 刘春芳 PANG Xiao-jun;LI Dong-wang;LIU Chun-fang(Department of Clinical Pharmacy,Second People’s Hospital of Qinzhou,Guangxi Qinzhou 535000,China;Pharmaceutical College,Guangxi Medical University,Guangxi Nanning 530021,China)

机构地区：[1]钦州市第二人民医院临床药学科,广西钦州535000 [2]广西医科大学药学院,广西南宁530021

出　　处：《中国医院药学杂志》2022年第13期1321-1325,1344,共6页Chinese Journal of Hospital Pharmacy

基　　金：钦州市新型冠状病毒感染的肺炎疫情应急科技攻关专项(编号:20204303)。

摘　　要：目的:通过蛋白水平和基因分子水平分别测定鞣花酸、连翘酯苷B、橙皮苷、连翘酯苷A、绿原酸、槲皮素、黄芩苷、连翘苷、苦参碱对血管紧张素转化酶2(angiotensin converting enzyme 2,ACE2)表达的影响,初步筛选出具有抗新型冠状病毒受体ACE2活性的单体化合物,为从中药中研发具有抗新型冠状病毒的活性化合物提供依据。方法:以具有人Ⅱ型肺泡细胞(ATⅡ)特征的A549细胞为实验模型,分别采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)法、蛋白免疫印迹法(Western blot,WB)、实时荧光定量PCR(real-time fluorescent quantitative polymerase chain reaction,qRT-PCR)法检测鞣花酸、连翘酯苷B、橙皮苷、连翘酯苷A、绿原酸、槲皮素、黄芩苷、连翘苷、苦参碱对A549细胞增殖及ACE2蛋白表达、ACE2 mRNA表达的影响。结果:实验检测到上述药物对A549细胞的增殖有一定的抑制作用(P<0.05),且呈现出浓度依赖性和时间依赖性;同时能够下调ACE2蛋白及其mRNA的表达(P<0.05)。结论:筛选出的鞣花酸、连翘酯苷B、橙皮苷、连翘酯苷A、绿原酸、槲皮素、黄芩苷、连翘苷、苦参碱能通过调节A549细胞的增殖、下调ACE2蛋白及其mRNA的表达,发挥潜在抗新型冠状病毒活性作用。OBJECTIVE To detect the effects of ellagic acid,forsythoside B,hesperidin,forsythiaside A,chlorogenic acid,quercetin,baicalin,phillygenin and matrine on ACE2 expression at protein level and gene molecular level,so as to preliminarily screen out mono-compounds with anti-novel coronavirus receptor ACE2 activity,and to provide basis for the development of novel coronavirus resistant active compounds from traditional Chinese medicine.METHODS A549 cells with the characteristics of human typeⅡalveolar cells(ATⅡ)were used as experimental model.The effects of ellagic acid,forsythoside B,hesperidin,forsythiaside A,chlorogenic acid,quercetin,baicalin,phillygenin and matrine on the proliferation,expression of ACE2 protein and ACE2 mRNA of A549 cells were detected respectively by MTT assay,Western blot and real-time quantitative PCR.RESULTS It was detected that the above drugs had a certain inhibitory effect on the proliferation of A549 cells(P<0.05),and the concentration dependence and time dependence were observed.Moreover,the drugs could down-regulate the expression of ACE2 protein and its mRNA(P<0.05).CONCLUSION The selected ellagic acid,forsythoside B,hesperidin,forsythiaside A,chlorogenic acid,quercetin,baicalin,phillygenin and matrine can exert potential anti-novel coronavirus activity by regulating the proliferation of A549 cells and down-regulating the expression of ACE2 protein and its mRNA.

关 键 词：新型冠状病毒 血管紧张素转化酶2 中药活性 噻唑蓝 免疫印迹法 

分 类 号：R285.5[医药卫生—中药学]