7种抗疟药抑制疟色素形成及其体外、体内抗日本血吸虫作用的比较观察  

Comparative Observation on Inhibition of Hemozoin Formation and Their in vitro and in vivo Antischistosome Activity Displayed by 7 Antimalarial Drugs

作　　者：薛剑[1] 姜斌[1] 刘丛珊[1] 孙军 肖树华[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所，卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾、血吸虫病和丝虫病合作中心,上海200025 [2]同济大学传染病与疫苗研究所,同济大学医学院,上海200092

出　　处：《中国寄生虫学与寄生虫病杂志》2013年第3期161-169,共9页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家国际科技合作专项(No.2010DFB73280)~~

摘　　要：目的比较、观察7种抗疟药抑制疟色素(hemozoin)的形成,与其体外和体内抗血吸虫的作用。方法抑制疟色素形成是通过观察25μmol/L磷酸氯喹、盐酸奎宁、奎尼丁、盐酸甲氟喹、磷酸咯萘啶和苯芴醇,以及100μmol/L蒿甲醚在pH 4.0～5.0的乙酸钠-高铁血红素(hematin)溶液中对β-hematin形成的抑制作用。用含10%小牛血清的RPMI1640培养基培养日本血吸虫成虫,测定上述7种抗疟药的半数和95%致死浓度(LC50和LC95)。观察奎宁、氯喹和咯萘啶伍用氯化血红素对体外培养血吸虫的致死作用,以及7种抗疟药口服或腹腔注射对感染日本血吸虫成虫小鼠的疗效。结果25μmol/L咯萘啶对pH 4.4～5.0的乙酸钠溶液中的hematin具有明显抑制β-hematin形成的作用,抑制率为81.3%～97.0%。在pH为4.6的乙酸钠-hematin溶液中,25μmol/L甲氟喹、氯喹或奎宁对β-hematin形成的抑制率分别为79.7%、72.8%和65.8%;在pH为4.8～5.0的乙酸钠-hematin溶液中,上述3种药物明显抑制β-hematin的形成,抑制率分别为83.1%～90.6%、41.9%～49.0%和53.2%～62.0%。25μmol/L本芴醇在pH为4.6、4.8和5.0的乙酸钠-hematin溶液中分别有74.3%和40.4%～40.5%的β-hematin形成抑制率。在相同浓度下,奎尼丁在pH为4.8和5.0的乙酸钠-hematin溶液中对β-hematin形成的抑制率分别为53.4%和50.9%,而100μmol/L蒿甲醚对pH 4.4～4.8的乙酸钠-hematin溶液中的β-hematin形成仅有轻度抑制作用,抑制率为16.6%～25.0%。甲氟喹、咯萘啶、奎宁和奎尼丁对体外培养血吸虫的LC50和LC95分别为4.93和6.123μg/ml,37.278和75.703μg/ml,93.688和134.578μg/ml,101.534和129.957μg/ml;而血吸虫在含100或120μg/ml氯喹、本芴醇和蒿甲醚的培养液中培养3 d,无或仅少数虫体死亡。用对血吸虫无效的奎宁50μmol/L(20μg/ml)和氯喹50μmol/L(26μg/ml)与氯化血红素153.4μmol/L(100μg/ml)伍用培养血吸虫,前者在培养的1～3 d内全部虫体死亡,而氯喹与氯化血红素伍�Objective To observe and compare the inhibition of hemozoin formation and the in vitro as well as in vivo antischistosomal activity induced by seven antimalarial drugs. Methods Inhibition of hemozoin formation displayed by chloroquine phosphate, quinine hydrochloride, quinidine, mefloquine hydrochloride, pyronaridine phosphateand lumefantrine at 25 μmol/L, and artemether at 100 μmol/L was performed by assay of inhibition of β-hematin formation in 1 mol/L sodium acetate buffers containing hematin with various pH of 4.0, 4.2, 4.4, 4.6, 4.8, and 5.0. In in vitro antischistosomal study, the medium of RPMI 1640 supplemented by 10% calf serum was used to maintain the adult Schistosoma japonicum, and the 50% and 95% lethal coneentratrions（LC50 and LC95） to kill the adult worms of each drug were then determined. Meanwhile, the interaction of quinine, pyronaridine and chloroquine combined with hemin against adult schistosomes was also undertaken. As to in vivo test, the efficacy of seven antimalarial drugs administered orally or intraperitoneally to mice infected with adult sehistosomes was observed. Results In the acidic acetate-hematin solution, 25 μmol/L pyronaridine showed significant inhibition of β-hematin formation at pH 4.4-5.0 with inhibition rates of 81.3%- 97.0%. At pH 4.6, the inhibition rates of β-hematin formation in aeetate-hematin solution induced by mefloquine, chloroquine or quinine at concentration of 25 μmol/L were 79.7%, 72.8% or 65.8%, respectively, and the 13-hematin formation was continually inhibited by these 3 antimalarial drugs at pH 4.8 and 5.0 with inhibition rates of 83.1%- 90.6%, 41.9%-49.0% or 53.2-62.0%. The inhibition rates of 13-hematin formation at pH 4.6 and 4.8-5.0 induced by lumefantrine 25 μmol/L were 74.3% and 40.4%-40.5%, respectively. While under the same concentration of quinidine, 53.4% and 50.9% inhibition rates of 13-hematin formation were observed at pH 4.8 and 5.0. As to artemether, higher concentration of 100 Ixmol/L only showed light inhibition of [3-hematin f

关 键 词：日本血吸虫 抗血吸虫作用 抗疟药 疟色素 氯化血红素 体外试验 体内试验 

分 类 号：R383.24[医药卫生—医学寄生虫学]