去氢骆驼蓬碱衍生物DH-330体外对细粒棘球呦线粒体凋亡通路中Cyt-C和Caspase-3的影响

Effects of the harmine derivative DH-330 on Cyt-C and C aspase-3 in the mitochondrial apoptotic pathway of Echinococcus granulosus in vitro

目的探讨去氢骆驼蓬碱衍生物DH-330体外诱导对细粒棘球蚴线粒体调亡通路中Cyt-C和Caspase-3的影响。方法体外培养细粒棘球喲,随机分为空白对照组、溶剂对照组(终体积浓度为1%DMSO)、阳性对照组(阿苯达唑亚砜组,ABZSO,培养液中终浓度为6.25、12.50、25.00、50.00、100.00 pug/L)及DH-330处理组(培养液中终质量浓度为6.25、12.50、25.00、50.00、100.00ug/L).连续用药6d.染色观察各组细粒棘球蚴存活率,通过酶联免疫吸附试验检测评价DH-330对细粒棟球蚴线粒体Cyt-C释放量,Caspase-3酶活性的影响,绘制活力曲线。结果DH-330干预后对细粒棘球喲形态产生显著影响,50.100 yμg/L DH-330处理3 d和6 d细粒棘球蚴存活率分别为59.33%、43.25%和21.45%、3.25%,DH-330浓度组(培养液中终质量浓度为12.50、25.00、50.00、100.00 ug/L)对细粒棘球蚴的杀伤作用呈时间、浓度依赖性。作用6 d,ED50值为25.44 pug/L。给予不同剂量的DH 330干预后Caspase 3酶活性显著增高(P<0.01).且呈浓度依赖性与时间依赖性;作用3 d时随着药物浓度的增加,细粒棘球蚴Cy-C表达水平逐渐增高(P<0.05).6 d时随着药物浓度的增加,细粒棘球蚴Cy+-C表达水平逐渐降低(P<0.05)。结论药物干预过程中,Caspase-3酶活性和Cyt-C释放量显著增加,表明线粒体凋亡通路可能参与DH-330诱导的细粒棘球蚴凋亡过程,具体机制有待进一步研究。

Objective To investigate the effect of the harmine derivative DH-330 on Cyt-C and C aspase-3 in the mito-chondrial apoptotice pathway of Echinococcus granulosus in vitro.Methods E.granulosus was cultured in vitro.E,gramulosus were randomly divided into a blank control group,a solvent control group(DMSO with 1%final volume con-centration),a positive control group(albendazole sulfoxide(ABZS0)group;the final mass concentration of culture me-dium was 6.25.12.50.25.00,50.00.100.00μug/L).and a DH-330 treatment group(the final mass concentration in culture medium was 6.25,12.50.25.00,50.00,or 100.00 ug/L).The drug was coninuously administered for6 d.and the survival rate of Echinococcus granulusus was observed.The effect of DH-330 on the release of Cyt-C by and the C aspase-3 activity of mitochondrin of E.granulosus was evaluated using an enzyme-linked immunosorbent assny,and a vi-ability curve was plotted.Results The morphology of E.granulosus was significantly affected by DH-330 interven-tion.When E.gramulosus was treated with 50 or 100 pμg/L DH 330 for 3d.it had a survival rate of 59.33%or 43.25%;when treated for 6 d.it had a survival rateof 21.45%or 3.25%.In the DH-330 concentration group(the final mass con-centration in culture medium was 12.50,25.00.50.00,or 100.00 pug/1).the klling of E.granulosts was time-and concentration-dependent.After 6 d.the ED。value was 25.44 pug/L.Caspase-3 activity increased significantly after ad-ministration of different doses of DH-330(P<0.01).and that action was concentration-and time dependent.I he level of expression of Cy1-C by E.granulosus increased gradually with an increase in the drug concentrationat 3d(P<O.05).At6d,the level of expressionof CytC by E.granulosus decreased gradually with an increase in the drug concentration(P<0.05).Conclusion During a drug intervention.the activity of Caspase-3 and the release of Cyt C increased signif-cantly,indicating that the mitochondrial apoptosis pathway may be involved in DH-330-induced apoptosis of E.granulo-sus.The specific mechanism needs to be studied further.