雄黄对细粒棘球蚴原头节体外生长及抗氧化酶活性的影响

目的初步探讨雄黄对体外细粒棘球蚴原头节生长及抗氧化酶的影响。方法在体外培养的基础上,将不同浓度雄黄分别作用细粒棘球蚴原头节2 d,光镜下观察原头节活力及形态变化;扫描电镜(SEM)和透射电镜(TEM)观察原头节表面及内部超微结构改变;采用ELISA法测定SOD、ROS、HO-1和NQO-1表达情况;采用Caspase-3试剂盒检测原头节Caspase-3酶活性。结果250、500、1000、2000μmol/L雄黄体外作用于细粒棘球蚴原头节均能抑制其生长,雄黄作用2 d后光镜下观察原头节形态结构均发生改变,虫体萎缩,伊红染色呈红色(正常虫体为透明无色);SEM下观察原头节虫体皱缩,原头节正常形态被破坏;TEM下观察原头节内部微绒毛减少,合胞体带变薄、结构松散并有少量脂滴。EUSA检测SOD、HO-1和NQO-1活性均呈下降趋势,ROS和caspase-3酶活性均呈升高趋势(均P<0.05)。结论雄黄体外可抑制细粒棘球蚴原头节生长,破坏原头节形态结构,降低抗氧化酶活性,该抑制作用与机体抗氧化防御系统平衡失调有关。

In vitro effects of realgar on the growth and antioxidant enzyme activity of Echinococcus granulosus protoscoleces

Objectives To investigate the effect of realgar on the growth and antioxidant enzymes of Echinococcus larvae in vitro.Methods On the basis of an in vitro culture,Echinococcus larvae were treated with different concentrations of realgar for 3 days,and the viability of and morphological changes in Echinococcus larvae were observed under a light microscope.Internal microstructural changes were observed under SEM and TEM.ELISA was used to detect the expression of SOD,ROS,HO-1,and NQO-1 in the Echinococcus larvae.Caspase-3 assay kit was used to detect the activity of Caspase-3 enzyme in the Echinococcus larvae.Results Treatment with 250,500,1,000,or 2,000 mol/L realgar inhibited larval growth in vitro.After 6 d,the mortality rate of Echinococcus larvae was 63%when exposed to 1,000μmol/L realgar and 100%when exposed to 2,000μmol/L realgar.Eosin staining was observed in the medium concentration group after 2 d.The normal morphology of Echinococcus larvae was destroyed and the internal morphology and structure of Echinococcus larvae were severely damaged when observed under SEM and TEM.ELISA revealed a decrease in SOD,HO-1,and NQO-1 activity.The most significant decrease was noted in the 2,000 mol/L realgar group,and an increase in ROS and caspase-3 enzyme activity was noted(P<0.05 for all).Conclusion Realgar inhibits the growth of Echinococcus larvae in vitro,destroying their morphology and structure.It also reduces the activity of antioxidant enzymes,which is related to an imbalance in the body’s antioxidant defense system.