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目的:通过气相色谱-质谱联用(GC-MS)技术分析Hep-2细胞内源性代谢产物的相对含量变化,阐明金欣口服液体外抗RSV感染的作用机制。方法:RSV感染接种于6孔板上的Hep-2细胞,予金欣口服液治疗,24 h后予以液氮淬灭细胞,先后加入水-甲醇-氯仿(v:v:v,1:8.1:0.9)以便取少量细胞悬浮液行蛋白定量和萃取细胞代谢物,运用GC-MS检测细胞样品内源性代谢物及其相对含量的变化,并分析其可能参与的代谢通路。结果:RSV感染的细胞样本中共测得29种内源性代谢产物,其中谷氨酰胺、γ-氨基丁酸、甘氨酸、柠檬酸、乳酸等为差异代谢产物(P<0.05),主要与谷氨酰胺、甘氨酸、丙酮酸代谢通路有关。结论:金欣口服液体外治疗RSV感染的机制可能与调节RSV感染后引起的代谢紊乱有关。
OBJECTIVE: To analyze the relative content of endogenous metabolites in Hep-2 cells by gas chromatography-mass spectrometry (GC-MS) and elucidate the mechanism of anti-RSV infection of Jinxin oral liquid in vitro. Methods: Hep-2 cells inoculated on 6-well plates were infected with RSV and treated with Jinxin oral solution. After 24 h, cells were treated with liquid nitrogen to quench the cells. Water and methanol-chloroform (v: v: v, : 0.9) in order to quantify and extract cell metabolites by using a small amount of cell suspension protein. The changes of endogenous metabolites and their relative contents in cell samples were detected by GC-MS, and the possible metabolic pathways were analyzed. Results: A total of 29 endogenous metabolites were detected in the cell samples infected with RSV, among which glutamine, γ-aminobutyric acid, glycine, citric acid and lactate were the differential metabolites (P <0.05) Amide, glycine, pyruvate metabolic pathway related. Conclusion: The mechanism of Jinxin oral liquid in treating RSV infection may be related to the metabolic disorder caused by RSV infection.