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通过观察半枝莲总黄酮对副流感病毒1型(PIV-1)感染后宿主细胞膜电位、膜Na+-K+-ATP酶活性和膜流动性的影响,为揭示半枝莲总黄酮抗病毒作用机制提供实验依据。本研究采用流式细胞仪检测膜电位,定磷法检测Na+-K+-ATP酶活性,荧光漂白恢复法检测膜流动性。结果显示,PIV-1感染后宿主细胞膜电位下降,处于超极化状态,荧光强度为42.81±5.65(P<0.05);膜Na+-K+-ATP酶活性显著增加,达(11.01±3.93)μmolPi·mg-1(protein)·h-1(P<0.05);扩散系数显著下降为(0.41±0.15)×10-9cm2·s-1(P<0.05),荧光恢复率为19.85%,明显低于正常对照组。半枝莲总黄酮(3mg·mL-1)作用后,扩散系数和荧光恢复率均明显升高,分别为(0.73±0.10)×10-9cm2·s-1和54.56%,与病毒对照组比较有显著性差异;而对宿主细胞膜的超极化状态和Na+-K+-ATP酶活性没有明显影响。实验结果表明,PIV-1感染后膜电位、Na+-K+-ATP酶活性和膜流动性等细胞膜能态和功能的改变,可能为病毒感染的细胞学机制之一;半枝莲总黄酮可能是通过改善细胞膜流动性,维持细胞膜的正常功能来发挥抗病毒感染的作用,而与膜电位和膜Na+-K+-ATP酶活性等能态来源的环节无关。
The objective of this study was to investigate the effects of Scutellaria barbatae flavonoids on the membrane potential, membrane Na + -K + -ATPase activity and membrane fluidity of host parainfluenza virus type 1 (PIV-1) Provide experimental basis. In this study, the membrane potential was measured by flow cytometry, the Na + -K + -ATPase activity was detected by fixed phosphorus assay, and the membrane fluidity was measured by fluorescence bleaching recovery assay. The results showed that the membrane potential of host cells decreased after PIV-1 infection and was hyperpolarised with a fluorescence intensity of 42.81 ± 5.65 (P <0.05), and the membrane Na + -K + -ATPase activity was significantly increased (11.01 ± 3.93) μmol Pi- (0.41 ± 0.15) × 10-9cm2 · s-1 (P <0.05), and the fluorescence recovery rate was 19.85%, which was significantly lower than Normal control group. Diffusion coefficient and fluorescence recovery rate of Scutellaria barbata total flavonoids (3mg · mL-1) were significantly increased (0.73 ± 0.10) × 10-9cm2 · s-1 and 54.56%, respectively, compared with the virus control group There is a significant difference; but the host cell membrane hyperpolarization state and Na + -K + -ATPase activity had no significant effect. The experimental results showed that the change of membrane potential and function of membrane potential, Na + -K + -ATPase activity and membrane fluidity after PIV-1 infection may be one of the cytological mechanisms of virus infection; Scutellaria barbata flavonoids may be By improving the cell membrane fluidity and maintaining the normal function of the cell membrane to play an anti-viral infection role, and membrane potential and membrane Na + -K + -ATP enzyme activity and other energy sources have nothing to do.