新冠肺脾气虚方对体外新冠病毒增殖及炎症因子表达的影响  

The effects of the recommended prescription for COVID-2019-lung-spleen qi deficiency on in vitro SARS-CoV-2 replication and pro-inflammatory factors expression

作　　者：谢佩芳 李慧[2] 马钦海 李洪梅[1] 李芳[3] 邵榆岚 方越 沈智丽 李蓉涛 杨子峰[3] 赵金存 董书维 杨洪军[2] 夏雪山[1] XIE Pei-fang;LI Hui;MA Qin-hai;LI Hong-mei;LI Fang;SHAO Yu-lan;FANG Yue;SHEN Zhi-li;LI Rong-tao;YANG Zi-feng;ZHAO Jin-cun;DONG Shu-wei;YANG Hong-jun;XIA Xue-shan(Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China;Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;State Key Laboratory of Respiratory Disease of Guangzhou Medical University, Guangzhou 510120, China)

机构地区：[1]昆明理工大学生命科学与技术学院,云南昆明650500 [2]中国中医科学院中药研究所,北京100700 [3]广州医科大学呼吸疾病国家重点实验室,广州广东510120

出　　处：《中国药理学通报》2022年第3期460-469,共10页Chinese Pharmacological Bulletin

基　　金：中国中医科学院中医药防治流感技术体系项目(No ZZ13-035-04);云南省科技研发计划新冠应急项目(No 202003AC100006)。

摘　　要：目的研究新冠肺脾气虚方(简称4-1)对新冠病毒(SARS-CoV-2)及普通冠状病毒(HCoV-OC43,229E和NL-63)的体外抗病毒作用,探讨其抗病毒抗炎机理。方法通过LC-MS分析4-1化学药效物质,MTT法检测药物毒性,CPE法确定该处方体外抗病毒活性,qRT-PCR法检测病毒载量和炎症因子表达水平。结果4-1含黄酮、甾体、倍半萜等94种化合物,对新冠及3种普通冠状病毒的选择指数范围为(8.44±0.49)~(52.26±2.3)。该处方在10、5、2.5 g·L^(-1)剂量下可抑制新冠病毒增殖,影响ACE2及S蛋白mRNA表达,下调IL^(-1)α及CCL-5/RANTES过表达;在20、10、5 g·L^(-1)等剂量下可抑制3种普通冠状病毒增殖,且抑制OC43、229E引起的IL-6、CXCL-8/IL-8、CXCL^(-1)0/IP-10、TNF-α、IFN-α、CCL-2/MCP-1、CXCL-9/MIG和CCL-5/RANTES表达上调,且抑制作用均呈剂量依赖性。结论肺脾气虚方具有抗多种冠状病毒增殖的活性,可抑制冠状病毒诱导的炎症因子表达上调。该研究为常见呼吸道病毒感染和新冠病毒等新发传染病的中药治疗提供了研究依据。Aim To determine the antiviral and anti-inflammatory effects of the recommended prescription for COVID-2019-lung-spleen qi deficiency(4-1)against in vitro infection of SARS-CoV-2 and common coronaviruses.Methods The main chemical substances of 4-1 were analyzed by LC-MS.The toxicity and antiviral effects of of 4-1 were detected by MTT and by CPE assay,respectively.The viral loads in cell supernatant and the expression of inflammatory factors induced by viral infection were determined by qRT-PCR.Results The recommended prescription 4-1 contained 94 chemical compounds,including flavonoids,steroids,sesquiterpenoids,and so on.The range of selection indexes for SARS-CoV-2 and common coronaviruses was 8.44±0.49~52.26±2.3.This prescription could inhibit the proliferation of SARS-CoV-2,the expression of ACE2 and S mRNA,and down regulate IL^(-1)αand CCL-5/RANTES at 10,5,and 2.5 g·L^(-1) doses.Further,at doses of 20,10 and 5 g·L^(-1),it could inhibit the proliferation of three common coronaviruses and suppress the overexpression of IL-6,CXCL-8/IL-8,CXCL^(-1)0/IP-10,TNF-α,IFN-α,CCL-2/MCP-1,MIG and CCL-5/RANTES induced by OC43/229E infection.The inhibitory effects were dose-dependent.Conclusions The prescription 4-1 has antiviral and anti-inflammation effects against multiple coronaviruses.This study provides the research basis for the treatment of common respiratory viral infections and emerging infectious diseases such as COVID-19 by using traditional Chinese medicine.

关 键 词：新冠肺脾气虚推荐处方 4-1 新冠病毒 普通冠状病毒 抗病毒 抗炎 

分 类 号：R289.5[医药卫生—方剂学] R364.5[医药卫生—中药学]