SARS-CoV-2病毒E蛋白诱发A549细胞炎症反应研究

SARS-CoV-2 virus E protein induces inflammation in A549 cells

探讨SARS-CoV-2病毒E蛋白诱发人肺腺癌(A549)细胞炎症反应机制。采用原核表达技术制备并导入大肠杆菌BL21菌株中诱导表达,经Ni-NTA介质纯化获得重组E蛋白。分别以终浓度10μg·L^(-1)和100μg·L^(-1)的重组E蛋白作用于A549细胞1 h,通过荧光定量PCR法检测细胞NLRP3,Caspase^(-1)和IL^(-1)β基因表达量。SARS-CoV-2病毒E蛋白被成功表达纯化,能够被新型冠状病毒肺炎治愈者血清所识别,并显著诱导A549细胞NLRP3,Caspase^(-1)和IL^(-1)β基因表达上调。表明细胞NRLP3炎症复合体被激活,同时产生大量IL^(-1)β促炎细胞因子,并可能会造成严重组织损伤。

To study the inflammation of A549 cells induced by SARS-CoV-2 virus E protein,the recombinant E protein was prepared by prokaryotic expression system and expressed in E.coli BL21 strain.The recombinant E protein was purified by Ni-NTA.The A549 cells were treated with recombinant E protein at concentrations of 10μg·L^(-1) and 100μg·L^(-1) for 1 h respectively.The mRNA expressions of NLRP3,Caspase^(-1) and IL^(-1)βwere determined by fluorescence quantitative PCR.SARS-CoV-2 virus E protein was successfully expressed and could be recognized by antibodies in the serum of patients cured by COVID^(-1)9.The concentration of 10μg·L^(-1) and 100μg·L^(-1) recombinant E protein could significantly increased the mRNA expressions of NLRP3,Caspase^(-1) and IL^(-1)βin A549 cells.SARS-CoV-2 virus E protein can induce inflammation in A549 cells.The NRLP3 inflammasome signal pathway in A549 cells were activated,and a large amount of pro-inflammatory cytokines were produced,which is an important factor in inducing tissue inflammation.