人血清抗SARS-CoV-2感染代谢产物的筛选。a,b，人血清代谢产物在SARS-CoV-2感染中的作用。研究设计示意图(a).人血清源滤液孵育损伤SARS-CoV-2复制(b).阴性对照:用未感染SARS-CoV-2的细胞培养基孵育的细胞。对于上液组和下液组，每个圆点代表一个供体(n = 8个健康供体)。上层液体与下层液体P = 0.0001，细胞培养基与上层液体、细胞培养基与下层液体P < 0.0001。c,d，预防SARS-CoV-2感染的人血清代谢物的鉴定。筛选实验设计示意图(c).人血清代谢产物(s)在SARS-CoV-2感染中的作用(d).病毒RNA量归一化为人GAPDH。点是平均值。e，通过免疫荧光染色评价人血清中6种代谢物的抗病毒活性。核衣壳用Alexa Fluor 546标记的抗兔IgG(红色)染色。 The nuclei were stained with To-Pro-3 iodide (blue). The stained cells were examined using a Zeiss LSM 880 meta confocal microscope in multitrack mode (i). Representative of three confocal immunofluorescence images from three biological replicates was shown. Scale bars, 20 μm. (ii) Mean fluorescence intensity in differently treated cells. Three images stained with nucleocapsid and Alexa Fluor 546-conjugated anti-rabbit IgG individually selected from three biological replicates were used to determine the mean fluorescence intensity with ImageJ (National Institutes of Health). AU, arbitrary units. P < 0.0001 for vehicle versus 1,5-AG, vehicle versus 1-napthol, vehicle versus 4-HA, vehicle versus 5-MT, vehicle versus CDCA, vehicle versus ellagic acid. f,g, Measurement of the half maximal inhibitory concentration (IC50) of these candidate metabolites. Half maximal inhibitory concentrations of these metabolites (f). The viral loads in the cell supernatant were detected with a plaque-formation assay at 40 h post-infection. The gray dotted line represents the 50% inhibition ratio (n = 3 biological independent samples). Biological characterizations of candidate metabolic component(s) (g). Data are presented as the mean ± s.e.m. (b,e,f). Data were analyzed using a two-tailed Student's t-test. P values were adjusted using Dunnett's test. ***P < 0.001, ****P < 0.0001. Experiments were performed independently at least three biological replicates with comparable results. Credit:自然的新陈代谢(2022)。DOI: 10.1038 / s42255 - 022 - 00567 - z
