SARS-CoV-2 N Protein Induces Acute Lung Injury in Mice via NF-ĸB Activation.
SARS-CoV-2 N Protein Induces Acute Lung Injury in Mice via NF-ĸB Activation.
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SARS-CoV-2 N 蛋白通过 NF-äB 激活诱导小鼠急性肺损伤
DOI：
10.3389/fimmu.2021.791753
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发表时间：
2021
影响因子：
7.3
通讯作者：
Xu J
中科院分区：
文献类型：
--
作者：
Xia J;Tang W;Wang J;Lai D;Xu Q;Huang R;Hu Y;Gong X;Fan J;Shu Q;Xu J
研究背景SARS-CoV-2感染可引起急性呼吸道综合征。据报道，SARS-CoV-2核衣壳蛋白（N-蛋白）在感染期间早期出现在体液中。N蛋白直接参与肺损伤的机制尚不清楚。方法用脂多糖（LPS）中和剂多粘菌素B预处理重组N蛋白。C57 BL/6、C3 H/HeJ（对LPS有抗性）和C3 H/HeN（C3 H/HeJ的对照）小鼠通过气管内施用暴露于N-蛋白以检查急性肺损伤。在体外，将骨髓源性巨噬细胞（BMDM）与N-蛋白一起培养以研究核因子κ B（NF-κ B B）p65的磷酸化、巨噬细胞极化和促炎细胞因子的表达。结果N蛋白可引起C57 BL/6小鼠急性肺损伤，肺组织蛋白通透性、细胞总数、中性粒细胞浸润及炎性细胞因子水平升高。N-蛋白还引起C3 H/HeJ和C3 H/HeN小鼠的肺损伤，表明该作用不能归因于LPS污染。N-蛋白在体外触发NF-κ B p65的磷酸化，其通过N-蛋白变性和用N-蛋白的抗体处理而消除，证明该作用是N-蛋白特异性的。此外，N蛋白促进M1巨噬细胞极化和促炎细胞因子的表达，这也被N蛋白变性和N蛋白抗体阻断。此外，N-蛋白诱导肺中NF-κ B p65磷酸化，而NF-κ B抑制剂吡咯烷二硫代氨基甲酸酯减轻N-蛋白对急性肺损伤的作用。结论SARS-CoV-2 N蛋白本身具有毒性，可引起小鼠急性肺损伤。N-蛋白和NF-NFB通路都可能是治疗2019冠状病毒病（COVID-19）多器官损伤的治疗靶点。
Background Infection of SARS-CoV-2 may cause acute respiratory syndrome. It has been reported that SARS-CoV-2 nucleocapsid protein (N-protein) presents early in body fluids during infection. The direct involvement of N-protein in lung injury is poorly understood. Methods Recombinant N-protein was pretreated with polymyxin B, a lipopolysaccharide (LPS)-neutralizing agent. C57BL/6, C3H/HeJ (resistant to LPS), and C3H/HeN (control for C3H/HeJ) mice were exposed to N-protein via intratracheal administration to examine acute lung injury. In vitro, bone marrow–derived macrophages (BMDMs) were cultured with N-protein to study phosphorylation of nuclear factor kappa B (NF-ĸB) p65, macrophage polarization, and expression of proinflammatory cytokines. Results N-protein produced acute lung injury in C57BL/6 mice, with elevated protein permeability, total cell count, neutrophil infiltration, and proinflammatory cytokines in the bronchioalveolar lavage. N-protein also induced lung injury in both C3H/HeJ and C3H/HeN mice, indicating that the effect could not be attributed to the LPS contamination. N-protein triggered phosphorylation of NF-ĸB p65 in vitro, which was abolished by both N-protein denaturation and treatment with an antibody for N-protein, demonstrating that the effect is N-protein specific. In addition, N-protein promoted M1 macrophage polarization and the expression of proinflammatory cytokines, which was also blocked by N-protein denaturation and antibody for N-protein. Furthermore, N-protein induced NF-ĸB p65 phosphorylation in the lung, while pyrrolidine dithiocarbamate, an NF-ĸB inhibitor, alleviated the effect of N-protein on acute lung injury. Conclusions SARS-CoV-2 N-protein itself is toxic and induces acute lung injury in mice. Both N-protein and NF-ĸB pathway may be therapeutic targets for treating multi-organ injuries in Coronavirus disease 2019 (COVID-19).
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影响因子：
39.3
作者：
Liu T;Zhang L;Joo D;Sun SC
通讯作者：
Sun SC
DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
影响因子：
3.7
作者：
Liao, QJ;Ye, LB;Wu, ZH
通讯作者：
Wu, ZH
DOI：
10.3791/1941
发表时间：
2010-08-29
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Helms, My N;Torres-Gonzalez, Edilson;Rojas, Mauricio
通讯作者：
Rojas, Mauricio
影响因子：
5.4
作者：
Narayanan, K;Maeda, A;Makino, S
通讯作者：
Makino, S