植物多酚对5型腺病毒感染后宿主细胞膜流动性的影响

Effect of Plant Polyphenols on the Membrane Fluidity of 293a Cells Infected by Adenovirus-5

目的:观察两种植物多酚EGCG(表没食子儿茶素没食子酸酯)、单宁酸对5型腺病毒感染后宿主细胞膜流动性的影响,初步探讨植物多酚抑制病毒作用机制。方法:利用DPH为荧光探针,以荧光偏振法观察5型腺病毒感染对宿主细胞膜流动性的影响,EGCG、单宁酸在此过程中对宿主细胞膜流动性改变的干预作用,结合CPE法测定的EGCG,单宁酸抑制5型腺病毒感染药效试验结果,以Spearman秩相关计算EGCG,单宁酸对宿主细胞膜流动性干预作用与其抑制病毒感染作用间的关系。结果:腺病毒感染细胞可提高宿主细胞膜流动性,滴度为100TCID50病毒可使293A细胞膜流动性提高15.34%。EGCG、单宁酸可抑制宿主细胞膜流动性,高浓度下分别可使宿主细胞膜流动性下降32.61%、25.75%。在病毒吸附前、吸附后、吸附同时,以EGCG、单宁酸处理宿主细胞,均发现宿主细胞膜流动性的干预作用与其抑制病毒感染效果高度相关(相关系数均>0.900,P<0.01)。结论:5型腺病毒感染会提高宿主细胞膜的流动性,而EGCG、单宁酸在干预腺病毒感染过程中对宿主细胞膜流动性的干预作用可能是其抑制腺病毒感染的机制之一。

Objective： To analyze the effect of two kinds of plant polyphenols： EGCG（（-）-epigallocatechin gallate） and Tannins on the membrane fluidity of 293 A cells infected by adenovirus-5 and explore the mechanism of its antiviral activity. Methods： The membrane phospholipid of 293 A was labeled with DPH. The changes of membrane fluidity were examined by fluorescence polarization experiment. The anti-viral effect of EGCG and Tannins was examined by CPE assay. The correlation of these polyphenols on membrane fluidity and anti-viral effect was analyzed by Spearman rank correlation analysis. Results： The membrane fluidity of 293 a cells increased sigificantly after adenovirus-5 infection, with 15.34% decreased by 100TCID50 virus titer. When the cell treated with EGCG and Tannins, the membrane fluidity decreased sigificantly, in the case of EGCG induced 32.61% decrease of membrane fluidity with high-concentration（640 μg/ML） and Tanins induce 25.75%. The tested polyphenol was included in the periods of pre-infection,adsorption, and post-infection, the effect of these polyphenols on membrane fluidity was highly correlated with their anti-viral effect.（R〉0.900, P〈0.01）. Conclusion： The anti-viral capacity of EGCG and Tannins is due to their intervention effect on the membrane fluidity of 293 A cells infected by adenovirus-5, which may be one of the mechanisms explaning its anti-adenovirus activity.