p38MAPK抑制剂SB202190体外对细粒棘球蚴原头节的作用

In vitro effects of p38MAPK inhibitor SB202190 on Echinococcus granulosus protoscoleces

目的探讨p38MAPK抑制剂SB202190体外抑制细粒棘球蚴原头节生长的作用。方法将体外培养的细粒棘球蚴原头节分别加入12.5、25、50、100μmol/L的SB202190中体外孵育。利用伊红染色的方法,在光镜下观察原头节的活力变化。实验重复3次;扫描电子显微镜下(SEM)下观察SB202190作用后原头节表面超微结构改变;不同浓度SB202190作用24h后,半胱氨酸天冬氨酸蛋白酶-3(caspase-3)活性检测试剂盒检测caspase-3酶活性。结果 50μmol/L和100μmol/L的SB202190作用1d后,头节活力开始下降。作用14d后,100μmol/LSB202190组无存活的头节,50μmol/L SB202190组的头节活力仅为13.8%。超微结构显示100μmol/LSB202190作用6d后,原头节顶突外翻、变形,顶突界面缺损,吸盘变形,体表出现虫蛀样损害。不同浓度SB202190作用24h后,与正常对照组比较,SB202190高浓度组原头节的caspase-3表达明显增高。结论 p38MAPK抑制剂SB202190在体外有明显的抑制细粒棘球蚴原头节生长的作用。

The aim of this study was to investigate the in vitro efficacy of the p38MAPK inhibitor SB202190 against Echinococcus granulosus protoscoleces. Protoscolices of Echinococcus granulosus were incubated in vitro with SB202190 at concentrations of 12.5,25,50,100μmol/L, and the effects on protoscoleces viability were monitored by 0.1% eosin staining and light microscopic inspection. Each experiment was repeated three times. Ultrastructural effects of SB202190 on Echinococcus granulosus protoscoleces in vitro were determined with SEM.Caspase-3 activity of SB202190 on Echinococcus granulosus protoscoleces in vitro was detected with kit. Starting from day 1 of incubation in the presence of 50μmol/L and 100μmol/L SB202190,viability of protoscoleces started to decline depending on the concentration of the inhibitors. After 14 days of incubation in the presence of 100μmol/L of SB202190, no survival protoscoleces remained and only 13.8% survival protoscoleces were present after 14 days of treatment with 50μmol/L of SB202190. Results of vitality tests were consistent with the tissue damage observed at the ultrastructural level. The morphological changes included contraction of the soma region, formation of blebs on the tegument,rostellar disorganization.After 24h of incubation in the presence of SB20219 at different concentrations,compared to the control protoscoleces, the activity of caspase-3 was found to be higher in protoscoleces were incubated with SB202190.The outcomes obtained demonstrate the favorable in vitro effect of the SB202190 against E.granulosus protoscoleces.