While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endo...While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endocytosis. We have found that SARS-CoV enters cells viapH- and receptor-dependent endocytosis. Treatment of cells with either SARS-COV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 （ACE2）, the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic path- way inhibitors and dominant-negative Epsl5 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research.展开更多
新型冠状病毒感染疫情发展迅速,中药在这场防治疫情的战争中发挥着积极作用,已有大量确有疗效的方剂应用于临床。然而由于中药具有多成分、多靶点、多途径协同作用的特点,传统的抗病毒中药筛选研究受限于实验室条件和实验周期,极大地限...新型冠状病毒感染疫情发展迅速,中药在这场防治疫情的战争中发挥着积极作用,已有大量确有疗效的方剂应用于临床。然而由于中药具有多成分、多靶点、多途径协同作用的特点,传统的抗病毒中药筛选研究受限于实验室条件和实验周期,极大地限制了筛选速度。该文基于分子对接进行潜在抗新型冠状病毒中药的筛选,通过获取2019-nCoV冠状病毒3CL水解酶(3CL protease,Mpro)和冠状病毒木瓜样蛋白酶(papain-like protease,PLP)的晶体结构,对中药化学成分数据库(traditional Chinese medicine database,TCMD 2009)进行基于分子对接的虚拟筛选,选择打分值排名前100的成分作为潜在抗新型冠状病毒活性的中药成分,并对成分来源中药进行频数统计,挑选命中成分数目较多的中药作为潜在抗新型冠状病毒活性的中药。研究结果显示,基于Mpro靶点筛选获得潜在抗新型冠状病毒活性成分12322个,代表性活性成分有紫菀五肽A、川芎嗪、丹酚酸B等,筛选得到的中药为生姜、紫菀、三七、川芎、丹参、干姜、瞿麦、大黄、肉苁蓉等;基于PLP靶点筛选获得潜在抗新型冠状病毒活性成分11294个,代表性活性成分有姜酮酚、白果醇、阿魏酸等,筛选得到的中药为党参、羌活、生姜、白果、川芎、瓜蒌、白芍、补骨脂、苦参、三七、当归等。结合文献报道的基于分子对接筛选的血管紧张素转换酶2(angiotensin converting enzyme 2,ACE2)靶点抑制活性候选中药,该文在湖南省新型冠状病毒肺炎中医药诊疗方案的基础上,将Mpro抑制、PLP抑制和ACE2抑制候选中药针对该疫情的合理应用进行了探讨,分析结果表明,PLP抑制活性筛选获得的瓜蒌及ACE2筛选获得的贝母包含于桑贝止嗽散、小陷胸汤之中,有清化热痰之功效,主治痰热壅肺所致的痰多咳喘或燥痰犯肺、干咳少痰、咯痰不爽,可应用于病毒作用初期外邪闭肺所致的咳嗽微�展开更多
文摘While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endocytosis. We have found that SARS-CoV enters cells viapH- and receptor-dependent endocytosis. Treatment of cells with either SARS-COV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 （ACE2）, the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic path- way inhibitors and dominant-negative Epsl5 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research.
文摘新型冠状病毒感染疫情发展迅速,中药在这场防治疫情的战争中发挥着积极作用,已有大量确有疗效的方剂应用于临床。然而由于中药具有多成分、多靶点、多途径协同作用的特点,传统的抗病毒中药筛选研究受限于实验室条件和实验周期,极大地限制了筛选速度。该文基于分子对接进行潜在抗新型冠状病毒中药的筛选,通过获取2019-nCoV冠状病毒3CL水解酶(3CL protease,Mpro)和冠状病毒木瓜样蛋白酶(papain-like protease,PLP)的晶体结构,对中药化学成分数据库(traditional Chinese medicine database,TCMD 2009)进行基于分子对接的虚拟筛选,选择打分值排名前100的成分作为潜在抗新型冠状病毒活性的中药成分,并对成分来源中药进行频数统计,挑选命中成分数目较多的中药作为潜在抗新型冠状病毒活性的中药。研究结果显示,基于Mpro靶点筛选获得潜在抗新型冠状病毒活性成分12322个,代表性活性成分有紫菀五肽A、川芎嗪、丹酚酸B等,筛选得到的中药为生姜、紫菀、三七、川芎、丹参、干姜、瞿麦、大黄、肉苁蓉等;基于PLP靶点筛选获得潜在抗新型冠状病毒活性成分11294个,代表性活性成分有姜酮酚、白果醇、阿魏酸等,筛选得到的中药为党参、羌活、生姜、白果、川芎、瓜蒌、白芍、补骨脂、苦参、三七、当归等。结合文献报道的基于分子对接筛选的血管紧张素转换酶2(angiotensin converting enzyme 2,ACE2)靶点抑制活性候选中药,该文在湖南省新型冠状病毒肺炎中医药诊疗方案的基础上,将Mpro抑制、PLP抑制和ACE2抑制候选中药针对该疫情的合理应用进行了探讨,分析结果表明,PLP抑制活性筛选获得的瓜蒌及ACE2筛选获得的贝母包含于桑贝止嗽散、小陷胸汤之中,有清化热痰之功效,主治痰热壅肺所致的痰多咳喘或燥痰犯肺、干咳少痰、咯痰不爽,可应用于病毒作用初期外邪闭肺所致的咳嗽微�
