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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(polyphyllinⅡ,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Li Cl,Na HCO3,Na2SO4和PBS均显著抑制PPⅡ溶血(P<0.05),阻断剂NPPB和DIDIS显著促进PPⅡ溶血(P<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(P<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(P<0.01);预处理1 min的1 mg·L-1安定和100μmol·L-1DHEA能完全拮抗PPⅡ溶血(P<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。
This article aims to investigate the hemolysis of anion channel protein and glucose transporter 1-mediated polyphyllin II (PPII), and to reveal the mechanism of hemolysis of PPII in vitro. In this experiment, the in vitro hemolysis of PPⅡ was detected by spectrophotometry. The effects of anion channel-related solutions and blockers, glucose channel-related inhibitors and multi-target drug dehydroepiandrosterone and diazepam on PPⅡ hemolysis were designed The main contents; Scanning electron microscopy and transmission electron microscopy PP Ⅱ erythrocyte morphology. Electron microscopy showed severe degeneration of erythrocytes treated with PPⅡ, with a large number of spherocytic and spiny red blood cells and vesicles. PPⅡ hemolysis test showed that the anion salt exudate Li Cl, Na HCO3, Na2SO4 and PBS significantly inhibited the hemolysis of PPⅡ (P <0.05), and NPPB and DIDIS blocked the hemolysis of PPⅡ (P <0.01). Under certain concentration (P <0.05). Glucose channel inhibitors Cytochalasin B and Verapamil were able to antagonize PPⅡ hemolysis (P <0.01). Pretreatment for 1 min 1 mg · L-1 diazepam and 100 μmol·L-1 DHEA completely antagonized PPⅡ hemolysis (P <0.01). The above results indicated that the mechanism of hemolysis of PPⅡ in vitro may be that GLUT1 is the main competitive site, which leads to the increase of intracellular osmotic pressure and hemolysis of erythrocytes by changing the anion channel transport activity and conformation.