Hua Xiaoting, Zhang Quan, Xu Wei, Wang Xiaogang, Wang Fei, Zhao Ping, Xia Qingyou
Biological Science Research Center, Southwest University, Chongqing 400715, China.
State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei, Chongqing 400715, China.
Int J Mol Sci. 2021 Jul 10;22(14):7423. doi: 10.3390/ijms22147423.
nucleopolyhedrovirus (BmNPV) is a pathogen that causes great economic losses in sericulture. Many genes play a role in viral infection of silkworms, but silkworm metabolism in response to BmNPV infection is unknown. We studied BmE cells infected with BmNPV. We performed liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS)-based non-targeted metabolomics analysis of the cytosolic extract and identified 36, 76, 138, 101, 189, and 166 different molecules at 3, 6, 12, 24, 48, and 72 h post BmNPV infection (hpi) compared with 0 hpi. Compounds representing different areas of metabolism were increased in cells post BmNPV infection. These areas included purine metabolism, aminoacyl-tRNA biosynthesis, and ABC transporters. Glycerophosphocholine (GPC), 2-hydroxyadenine (2-OH-Ade), gamma-glutamylcysteine (γ-Glu-Cys), hydroxytolbutamide, and 5-pyridoxolactone glycerophosphocholine were continuously upregulated in BmE cells post BmNPV infection by heat map analysis. Only 5-pyridoxolactone was found to strongly inhibit the proliferation of BmNPV when it was used to treat BmE cells. Fewer infected cells were detected and the level of BmNPV DNA decreased with increasing 5-pyridoxolactone in a dose-dependent manner. The expression of BmNPV genes ie1, helicase, GP64, and VP39 in BmE cells treated with 5-pyridoxolactone were strongly inhibited in the BmNPV infection stage. This suggested that 5-pyridoxolactone may suppress the entry of BmNPV. The data in this study characterize the metabolism changes in BmNPV-infected cells. Further analysis of 5-pyridoxolactone, which is a robust antiviral molecule, may increase our understanding of antiviral immunity.
核型多角体病毒（BmNPV）是一种在养蚕业中造成巨大经济损失的病原体。许多基因在病毒感染家蚕的过程中发挥作用，但家蚕对BmNPV感染的代谢反应尚不清楚。我们研究了感染BmNPV的BmE细胞。我们对胞质提取物进行了基于液相色谱-串联质谱（LC-MS/MS）的非靶向代谢组学分析，并与感染前0小时相比，在感染BmNPV后3、6、12、24、48和72小时分别鉴定出36、76、138、101、189和166种不同的分子。BmNPV感染后，细胞中代表不同代谢区域的化合物增加。这些区域包括嘌呤代谢、氨酰-tRNA生物合成和ABC转运蛋白。通过热图分析发现，甘油磷酸胆碱（GPC）、2-羟基腺嘌呤（2-OH-Ade）、γ-谷氨酰半胱氨酸（γ-Glu-Cys）、羟基甲苯磺丁脲和5-吡哆醇内酯甘油磷酸胆碱在BmNPV感染后的BmE细胞中持续上调。当用5-吡哆醇内酯处理BmE细胞时，仅发现5-吡哆醇内酯强烈抑制BmNPV的增殖。检测到的感染细胞减少，并且BmNPV DNA水平随着5-吡哆醇内酯剂量的增加呈剂量依赖性下降。在BmNPV感染阶段，用5-吡哆醇内酯处理的BmE细胞中BmNPV基因ie1、解旋酶、GP64和VP39的表达受到强烈抑制。这表明5-吡哆醇内酯可能抑制BmNPV的进入。本研究中的数据表征了BmNPV感染细胞中的代谢变化。对作为一种强大抗病毒分子的5-吡哆醇内酯的进一步分析，可能会增进我们对抗病毒免疫的理解。
