双氢青蒿素体外对疟原虫感染人红细胞膜通透性的影响研究
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摘要
青蒿素类药物抗疟机制尚未明确,目前较主流的多种青蒿素抗疟机制学说提示,青蒿素类药物可能存在多方面多途径的抗疟机制;另一方面,疟原虫在体内的发病机制相对明确,其中关于疟疾发病的"红细胞膜疟原虫诱生阴离子通道(PSAC)"研究揭示,红细胞感染疟原虫后,红细胞膜可以选择性增高对疟原虫生长增殖所需的红细胞外糖醇、嘌呤和氨基酸等阴离子营养物质的透膜运输,采用阴离子通道抑制剂可以抑制红内期疟原虫对胞外营养物质的摄取,阻断疟原虫的发育增殖。该文研究双氢青蒿素(DHA)体外对人源性HB3疟原虫感染人红细胞膜通透性的影响,以提示青蒿素类药物是否可以通过抑制红细胞膜通透性,发挥阻止和杀灭疟原虫红内期生长增殖作用。实验采用5%山梨醇可以透过红细胞膜特异性杀灭红内期疟原虫的原理,观察体外HB3培养体系中施加DHA与否,山梨醇对红内期疟原虫的杀灭效果差异,考察DHA是否可以影响疟原虫感染红细胞膜的通透性;结果显示,10 nmol·L-1的DHA(疟原虫体外培养体系终浓度)预刺激30 min后,可以显著减弱山梨醇对红内期HB3疟原虫的杀灭作用,DHA有可能是通过抑制疟原虫感染红细胞膜的通透性,阻碍了山梨醇透过红细胞膜,从而减弱了山梨醇对红内期疟原虫的杀灭作用。
In view of the fact that the antimalarial effects of artemisinins are significant but the mechanism has not yet been clarified and there are many different opinions,it is possible that artemisinins can produce high anti-malarial efficacy through various mechanisms and multiple pathways. In addition,the researches on the pathogenesis of malaria " erythrocyte membrane plasmodial surface anion channel( PSAC) " in the past few years have provided more positive findings,which may confirm and discover the new antimalarial mechanism of artemisinins. This paper was as to study the effect of dihydroartemisinin( DHA) in vitro on erythrocyte membrane permeability of HB3 plasmodium infection,with using the mechanism of 5% sorbitol can be used to kill the Plasmodium falciparum in red blood cell membrane selectively,the effectual difference of sorbitol on the killing of P. falciparum with adding DHA or not was detected,so as to investigate whether DHA can affect the permeability of the erythrocyte membrane. Result showed that,Pre-stimulation with 10 nmol·L-1 DHA( the final concentration of plasmodium in vitro culture system) for 30 min could significantly decrease the killing effect of sorbitol on the HB3 plasmodium in the P. falciparum erythrocytic cycle,and DHA may inhibit the permeability of the erythrocyte membrane for preventing sorbitol through the red blood cell membrane,thereby reducing the killing effect of sorbitol on the P. falciparum.
In view of the fact that the antimalarial effects of artemisinins are significant but the mechanism has not yet been clarified and there are many different opinions,it is possible that artemisinins can produce high anti-malarial efficacy through various mechanisms and multiple pathways. In addition,the researches on the pathogenesis of malaria " erythrocyte membrane plasmodial surface anion channel( PSAC) " in the past few years have provided more positive findings,which may confirm and discover the new antimalarial mechanism of artemisinins. This paper was as to study the effect of dihydroartemisinin( DHA) in vitro on erythrocyte membrane permeability of HB3 plasmodium infection,with using the mechanism of 5% sorbitol can be used to kill the Plasmodium falciparum in red blood cell membrane selectively,the effectual difference of sorbitol on the killing of P. falciparum with adding DHA or not was detected,so as to investigate whether DHA can affect the permeability of the erythrocyte membrane. Result showed that,Pre-stimulation with 10 nmol·L-1 DHA( the final concentration of plasmodium in vitro culture system) for 30 min could significantly decrease the killing effect of sorbitol on the HB3 plasmodium in the P. falciparum erythrocytic cycle,and DHA may inhibit the permeability of the erythrocyte membrane for preventing sorbitol through the red blood cell membrane,thereby reducing the killing effect of sorbitol on the P. falciparum.
引文
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[18]孙辰,李坚,周兵.青蒿素类药物的作用机制:一个长久未决的基础研究挑战[J].中国科学:生命科学,2012,42(5),345.
[19]Li J,Zhou B.Biological actions of artemisinin:insights from medicinal chemistry studies[J].Molecules,2010,15:1378.
[20]叶祖光,李思迪.青蒿素类抗疟药作用机理研究述评[J].中国中医药信息杂志,2014,21(9):5.
[21]中医研究院中药研究所药理研究室.青蒿的药理研究[J].新医药学杂志,1979,8(1):23.
[22]Ginsburg H,Kutner S,Krugliak M,et al.Characterization of permeation pathways appearing in the host membrane of Plasmodium falciparum infected red blood cells[J].Mol Biochem Parasitol,1985,14(3):313.
[23]Kutner S,Breuer W V,Ginsburg H,et al.Characterization of permeation pathways in the plasma membrane of human erythrocytes infected with early stages of Plasmodium falciparum:association with parasite development[J].J Cell Physiol,1985,125(3):521.
[2]Tu Y.The discovery of artemisinin(qinghaosu)and gifts from Chinese medicine[J].Nat Med,2011,17:1217.
[3]青蒿素结构研究协作组.一种新型的倍半萜内酯——青蒿素[J].科学通报,1977,22(3):142.
[4]Wang J,Zhu X.The recent advances of chemical biology in artemisinin target and mechanism[J].Chin Sci Bull,2017,62(18):1973.
[5]Hamill O P.Patassium channel currents in human red blood cells[J].J Physiol,1981,319:97.
[6]Grygorcxyk R,Schwarx W,Passow H.Ca++-activated K+channel in human red cells[J].Biophys J,1984,45:693.
[7]Kirk K.Channels and transporters as drug targets in the plasmodium-infected erythrocyte[J].Acta Trop,2004,89:285.
[8]Sanjay A Desai.Why do malaria parasites increase host erythrocyte permeability?[J].Trends Parasitol,2014,30(3):151.
[9]David A Hill,Sanjay A Desai.Malaria parasite mutants with altered erythrocyte permeability:a new drug resistance mechanism and important molecular tool[J].Future Microbiol,2010,5:81.
[10]Ginsburg H,Kutner S,Krugliak M,et al.Characterization of permeation pathways appearing in the host membrane of Plasmodium falciparum infected red blood cells[J].Mol Biochem Parasitol,1985,14(3):313.
[11]Kang M,Lisk G,Hollingworth S,et al.Malaria parasites are rapidly killed by dantrolene derivatives specific for the plasmodial surface anion channel[J].Mol Pharmacol,2005,68(1):34.
[12]Staines H M,Rae C,Kirk K.Increased permeability of the malaria-infected erythrocyte to organic cations[J].Biochim Biophys Acta,2000,1463:88.
[13]Cohn J V,Alkhalil A,Wagner M A,et al.Extracellular lysines on the plasmodial surface anion channel involved in Na+exclusion[J].Mol Biochem Parasitol,2003,132:27.
[14]郝明明,杨照青.恶性疟原虫体外培养的同步化方法[J].中国热带医学,2012(12)9:1154.
[15]O'Neill P M,Barton V E,Ward S A.The molecular mechanism of action of artemisinin—the debate continues[J].Molecules,2010,15:1705.
[16]Ho W E,Peh H Y,Chan T K,et al.Artemisinins:pharmacological actions beyond anti-malarial[J].Pharmacol Ther,2014,142:126.
[17]Meshnick S R,Taylor T E,Kamchonwongpaisan S.Artemisinin and the antimalarial endoperoxides:from herbal remedy to targeted chemotherapy[J].Microbiol Rev,1996,60:301.
[18]孙辰,李坚,周兵.青蒿素类药物的作用机制:一个长久未决的基础研究挑战[J].中国科学:生命科学,2012,42(5),345.
[19]Li J,Zhou B.Biological actions of artemisinin:insights from medicinal chemistry studies[J].Molecules,2010,15:1378.
[20]叶祖光,李思迪.青蒿素类抗疟药作用机理研究述评[J].中国中医药信息杂志,2014,21(9):5.
[21]中医研究院中药研究所药理研究室.青蒿的药理研究[J].新医药学杂志,1979,8(1):23.
[22]Ginsburg H,Kutner S,Krugliak M,et al.Characterization of permeation pathways appearing in the host membrane of Plasmodium falciparum infected red blood cells[J].Mol Biochem Parasitol,1985,14(3):313.
[23]Kutner S,Breuer W V,Ginsburg H,et al.Characterization of permeation pathways in the plasma membrane of human erythrocytes infected with early stages of Plasmodium falciparum:association with parasite development[J].J Cell Physiol,1985,125(3):521.