半枝莲总黄酮抗副流感病毒的作用机制

The cytology mechanism of anti-parainfluenza virus infection of total flavone of Scutellaria barbata

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。

In order to research into the cytology mechanism of anti-virus action of total flavone of Scutellaria barbata （TFSB）, the effects of TFSB on host cells membrane potential, Na^＋-K^＋-ATPase activity and membrane fluidity after parainfluenza virus typel （PIV-1） infection were studied. The changes of membrane potential which was fluorescent labeled with DiBAC4（3） and its changes were measured by flow cytometer. Phosphorus determination method and spectrophotometry were used to measure the Na^＋-K^＋-ATPase activity of Hep-2 cells membrane after PIV-1 infection. Hep-2 cells membrane phospholipids were fluorescent labeled with NBD-C6-HPC and membrane fluidity was measured by confocal scanning laser microscope. The result demonstrated that post PIV-1 infection membrane potential decreased significantly and the membrane was in a state of hyperpolarization, Na^＋-K^＋-ATPase activity increased significantly and membrane fluidity decreased significantly. There was no apparent interfere effect of TFSB on the changes of membrane potential and Na^＋-K^＋ATPase activity after PIV-1 infection, while membrane fluidity improved significantly. It was indicated that the cytology mechanism of PIV-1 infection might be related to membrane hyperpolarization, Na^＋-K^＋-ATPase activity increase and membrane fluidity decrease. TFSB can improve membrane fluidity and prevent the infection by protecting the cell membrane. But it is possible that the anti-PIV-1 mechanisms of TFSB hadnothing to do with membrane potential and Na^＋-K^＋-ATPase activity.