摘要
目的建立过氧化氢(H_(2)O_(2))体外诱导细粒棘球绦虫囊泡氧化损伤模型。方法采用终浓度为0、1、2、5 mmol/L的活性氧清除剂-N-乙酰半胱氨酸(NAC)对细粒棘球绦虫囊泡预先干预2 h,再分别用终浓度为0、50、100、200、400μmol/L的H_(2)O_(2)处理囊泡0.5 h,用活性氧检测试剂盒对囊泡体内的活性氧含量进行检测;采用激光共聚焦对1%二甲基亚砜(DMSO)、50μmol/L H_(2)O_(2)、100μmol/L H_(2)O_(2)、200μmol/L H_(2)O_(2)、200μmol/L H_(2)O_(2)+5 mmol/L NAC和5 mmol/L NAC干预0.5 h的囊泡进行体内活性氧观察并拍照;干预囊泡4 d,每天对囊泡的塌陷率进行统计;采用电镜对干预4 d后的囊泡超微结构进行观察。结果随着H_(2)O_(2)浓度的升高,囊泡体内的活性氧含量逐渐升高;H_(2)O_(2)和NAC干预囊泡后囊泡塌陷率结果显示H_(2)O_(2)单独及H_(2)O_(2)与NAC联合干预囊泡均可导致囊泡的活性降低,而H_(2)O_(2)和NAC联合干预囊泡可明显改善H_(2)O_(2)单独对囊泡的塌陷率的降低;电镜对H_(2)O_(2)和NAC干预囊泡后的超微结构观察结果显示H_(2)O_(2)可导致囊泡体内异染色质的增多,而NAC可缓解H_(2)O_(2)对囊泡体内异染色质的增多;激光共聚焦结果与活性氧含量检测结果一致。结论采用200μmol/L H_(2)O_(2)可诱导囊泡产生氧化损伤作用,且5 mmol/L NAC可有效的抑制H_(2)O_(2)诱导的氧化损伤作用,H_(2)O_(2)诱导囊泡氧化损伤模型的建立为抗包虫病药物氧化损伤作用研究奠定基础。
Objective To establish the oxidative damage model of Echinococcus granulosus vesicles induced by hydrogen peroxide(H_(2)O_(2)).Methods N-acetylcysteine(NAC),an active oxygen scavenger with final concentration of 0,1,2 and 5 mmol/L,was used to pre-treat with vesicles for 2 hrs,then 0,50,100,200 and 400μmol/L H_(2)O_(2) were used to treat the vesicles for 0.5 hrs,and the active oxygen content in the vesicles was detected by the active oxygen detection kit.Vesicles were intervented with 1%DMSO,50μmol/L H_(2)O_(2),100μmol/L H_(2)O_(2),200μmol/L H_(2)O_(2),200μmol/L H_(2)O_(2)+5 mmol/L NAC and 5 mmol/L NAC.The active oxygen in vesicles was observed by laser confocal focusing and photographed;the collapse rate of vesicles was counted every day for 4 days;the ultrastructure of vesicles after 4-day intervention was observed by electron microscope.Results With the increase of H_(2)O_(2) concentration,the content of active oxygen in the vesicles gradually increased.The results showed that H_(2)O_(2) alone and H_(2)O_(2) combined with NAC could reduce the activity of vesicles,while H_(2)O_(2) combined with NAC intervention could significantly improve the collapse rate of vesicles compared with H_(2)O_(2) alone;the ultrastructural observation by electron microscope of H_(2)O_(2) and NAC after vesicle intervention showed that H_(2)O_(2) could lead to the increase of heterochromatin in vesicle body,but could alleviate the increase of heterochromatin in vesicle body by NAC,which were consistent with the results of laser confocal on active oxygen content.Conclusion The results show that the oxidative damage of vesicles could be induced by 200μmol/L H_(2)O_(2),and 5 mmol/L NAC can effectively inhibit the oxidative damage induced by H_(2)O_(2).The establishment of the oxidative damage model of vesicles induced by H_(2)O_(2) lays a foundation for the study of the oxidative damage of anti-echinococcosis drugs.
作者
文丽梅
巩月红
吕国栋
张嘉伟
王建华
WEN Limei;GONG Yuehong;LV Guodong;ZHANG Jiawei;WANG Jianhua(Department of Clinical Pharmacy,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054;School of Pharmacy,Xinjiang Medical University,Urumqi 830011;State Key Laboratory of Pathogenesis,Prevention,Treatment of Central Asian High Incidence Diseases,Urumqi 830011;the First Affiliated Hospital of Xinjiang Medical University,Institute of clinical medicine,Urumqi 830054,China)
出处
《新疆医科大学学报》
CAS
2021年第3期285-289,304,共6页
Journal of Xinjiang Medical University
基金
国家自然科学基金(81560607,81860666)
省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2018-5)。
关键词
过氧化氢
细粒棘球绦虫
氧化损伤
N-乙酰半胱氨酸
hydrogen peroxide(H2O2)
Echinococcus granulosus
oxidative damage
N-acetylcysteine(NAC)
