青蒿琥酯体外抗细粒棘球蚴活性氧对DNA作用机制影响的研究

发布时间：2018-07-09 11:51

本文选题：青蒿琥酯 + 细粒棘球蚴　；参考：《中国病原生物学杂志》2017年08期

【摘要】：目的了解索青蒿琥酯体外抗细粒棘球蚴氧化损伤机制。方法以二甲基亚砜(DMSO)为溶剂对照组,H2O2为活性氧(reactive oxygen species,ROS)对照组,阿苯达唑(albendazole,ABZ)和硝唑尼特(nitazoxanide,NTZ)为药效对照组,青蒿琥酯低剂量组(65μmol/L),青蒿琥酯中剂量组(130μmol/L)和青蒿琥酯高剂量组(325μmol/L)为实验组,采用1%伊红染色法检测实验组药物干预4d的细粒棘球蚴死亡率,并观察病理组织学和超微结构变化;采用双氢罗丹明123(DHR123)法检测细粒棘球绦虫细粒棘球蚴体内ROS含量动态变化;采用单细胞凝胶电泳法(彗星实验)以Olive尾距(OTM)作为DNA损伤指标对各组细粒棘球蚴的DNA损伤情况进行分析。同时设活性氧清除剂甘露醇(mannitol,man)与药物联合干预细粒棘球绦虫细粒棘球蚴试验组作比较。结果与甘露醇联合药物干预组比较,青蒿琥酯中剂量组与H_2O_2组细粒棘球蚴死亡率降低(χ~2分别=46.371和59.328,P0.05);病理学观察青蒿琥酯组细粒棘球蚴顶突小钩脱落,角质层、生发层结构均遭到破坏;透射电镜观察细粒棘球蚴生发层出现大量脂滴,并且有异染色质出现;在330min内,青蒿琥酯低、中和高剂量组细粒棘球蚴ROS含量升高程度均明显高于阿苯达唑组、硝唑尼特组,并具有一定的剂量依赖性;青蒿琥酯高剂量组彗星实验图像拖尾明显,证明细粒棘球蚴DNA受损严重;青蒿琥酯低、中、高剂量组与阿苯达唑组、硝唑尼特组相比,OTM值比较均具有显著性差异(t=9.065、13.134、7.845、9.400、12.251和7.877,P0.01)。结论青蒿琥酯抗细粒棘球蚴的作用机制可能通过产生ROS导致其DNA损伤所致,为青蒿琥酯抗包虫病作用靶点的筛选和抗包虫病新药的开发奠定了基础。
[Abstract]:Objective to investigate the mechanism of artesunate on oxidative injury of echinococcus granulosus in vitro. Methods dimethyl sulfoxide (DMSO) as solvent control group, H _ 2O _ 2 as active oxygen species (reactive oxygen speciesRos) control group, albendazoleoxole ABZ and nitazoxanide NTZ as control group were used as control group. Artesunate low dose group (65 渭 mol / L), middle dose group (130 渭 mol / L) and artesunate high dose group (325 渭 mol / L) were used as experimental groups. The mortality rate of Echinococcus granulosus in experimental group was detected by 1% eosinolus staining for 4 days, and histopathological and ultrastructural changes were observed. The dynamic changes of Ros content in Echinococcus granulosus were detected by dihydrorhodamine 123 (DHR123) method. Single cell gel electrophoresis (comet assay) using Olive tail distance (OTM) as DNA damage index was used to analyze DNA damage of each group of echinococcus granulosus. At the same time, the experimental group of Echinococcus granulosus (Echinococcus granulosus) treated with mannitol (mannitolman) and drugs were compared. Results compared with the mannitol combined drug intervention group, the mortality rate of artesunate in middle dose group and S _ 2O _ 2 group was lower (蠂 ~ 2 = 46.371 and 59.328 P 0.05, respectively), and artesunate group was observed by pathology to observe the exfoliation of the parietal uncinate, the keratinocyte of artesunate group, and the relation between artesunate group and S _ 2O _ 2 group (P < 0.05). The structure of germinal layer was destroyed. A large number of lipid droplets and heterochromatin were observed in the germinal layer of Echinococcus granulosus by transmission electron microscope, and artesunate was low in 330min. The Ros content of Echinococcus granulosus in middle and high dose groups was significantly higher than that in albendazole group and niazonide group in a dose-dependent manner. The results showed that DNA damage of Echinococcus granulosus was serious and the OTM values of artesunate, albendazole and niazonide groups were significantly higher than those of albendazole group and niazonide group. Conclusion the mechanism of artesunate against echinococcus granulosus may be caused by DNA damage caused by Ros, which may lay a foundation for the screening of anti-hydatid targets of artesunate and the development of new anti-hydatid drugs.
【作者单位】：新疆医科大学药学院;新疆医科大学第一附属医院药学部临床药学科;新疆医科大学第一附属医院临床医学研究院新疆重大疾病医学重点实验室-省部共建国家重点实验室培育基地;
【基金】：国家自然科学基金项目(No.81560607,81360251) 新疆重大疾病医学重点实验室开放课题(No.SKLIB-XJMDR-2012-2) 新疆维吾尔自治区包虫病基础医学重点实验室开放课题(No.XJDX0202-2013-10)
【分类号】：R383.3

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