重楼皂苷Ⅱ溶血作用机制研究.docVIP

重楼皂苷Ⅱ溶血作用机制研究 　　[摘要] 该文旨在探讨阴离子通道蛋白和葡萄糖转运蛋白1介导的重楼皂苷Ⅱ（polyphyllin Ⅱ，PPⅡ）的溶血作用，初步揭示PPⅡ体外溶血作用机制。实验采用分光光度法检测PPⅡ体外溶血，设计了与阴离子通道相关的溶液与阻断剂、葡萄糖通道相关的抑制剂和多靶点药物脱氢表雄酮与安定对PPⅡ溶血的影响3个方面的主要内容;通过扫描电镜和透射电镜观察PPⅡ对红细胞形态的影响。电镜观察显示PPⅡ处理的红细胞变形严重，出现大量球形红细胞和棘形红细胞以及囊泡。PPⅡ体外溶血试验结果显示，阴离子盐渗液LiCl，NaHCO3，Na2SO4和PBS均显著抑制PPⅡ溶血（Plt;0.05），阻断剂NPPB和DIDIS显著促进PPⅡ溶血（Plt;0.01）;一定浓度下的高渗液氯化钠、果糖和葡萄糖显著拮抗PPⅡ溶血（Plt;0.05）;葡萄糖通道抑制剂细胞松弛素B和维拉帕米能显著拮抗PPⅡ溶血（Plt;0.01）;预处理1 min的1 mg?Llt;supgt;-1lt;/supgt;安定和100 μmol?Llt;supgt;-1lt;/supgt; DHEA能完全拮抗PPⅡ溶血（Plt;0.01）。以上结果表明PPⅡ体外溶血的机制可能是以GLUT1为主要的竞争性作用位点，通过改变阴离子通道转运活性和构象，最终引起胞内渗透压升高，红细胞破裂溶血。 　　[关键词] 阴离子通道蛋白;葡萄糖转运体1;重楼皂苷Ⅱ;溶血;红细胞;机制 　　Study on hemolytic mechanism of polyphyllin Ⅱ 　　NING Li-hua， ZHOU Bo， ZHANG Yao-xiang， LI Xin-ping* 　　（School of Veterinary Medicine， Northwest Agriculture and Forest University， Yangling 712100， China） 　　[Abstract] To study the hemolytic effect of polyphyllin Ⅱ（PP Ⅱ） mediated by anion channel protein and glucose transporter 1 （GLUT1）， in order to initially reveal its hemolytic mechanism in vitro. In the experiment， the spectrophotometric method was adopted to detect the hemolysis of PP Ⅱ in vitro and the effect of anion channel-related solution and blocker， glucose channel-related inhibitor and multi-target drugs dehydroepiandrosterone （DHEA） and diazepam on the hemolysis of PP Ⅱ. The scanning electron microscope and transmission electron microscope were used to observe the effect of PP Ⅱ on erythrocyte （RBC） morphology. The results showed that PP Ⅱ-processed blood cells were severely deformed into spherocytes， acanthocyturia and vesicae. According to the results of the PP Ⅱ hemolysis experiment in vitro， the anion hypertonic solution LiCl， NaHCO3， Na2SO4 and PBS significantly inhibited the hemolysis induced by PPⅡ （Plt;0.05）， while blockers NPPB and DIDS remarkably promoted it （Plt;0.01）. Hyperosmotic sodium chloride， fructose and glucose at specific concentrations notably antagonized the hemolysis induced by PP Ⅱ （Plt;0.05）. The glucose cha