甘氨鹅脱氧胆酸对体外细粒棘球蚴原头节活力及ROS影响

目的 研究甘氨鹅脱氧胆酸(glycochenodeoxycholic acid, GCDCA)对体外细粒棘球蚴原头节活力及活性氧(Reactive oxygen species,ROS)水平的影响。方法 将细粒棘球蚴原头节分为对照组及GCDCA处理组(500、1 500、2 500 μ mol/ L),应用体外培养技术,通过伊红染色法每天定时在光学显微镜下观察原头节的活力并记录存活和死亡数目。使用试剂盒检测不同浓度GCDCA处理 24 h后各组原头节体内caspase-3酶活性变化;DCFH- DA染色法荧光显微镜观察 GCDCA作用24 h后原头节内 ROS水平变化。结果 500、1 500、2 500 μ mol/ L GCDCA处理后均可导致原头节活力减弱,且最高浓度组在第7 d时原头节全部死亡。500、1 500、2 500 μ mol/ L GCDCA组作用原头节24 h后 caspase-3酶活性与对照组相比明显升高(F=555.162, P<0.05)。500、1 500、2 500 μ mol/ L GCDCA作用原头节24 h后原头节内 ROS水平与对照组相比明显升高(F=216.901, P<0.05)。结论 GCDCA 能抑制体外原头节生长,促使原头节死亡,可能与活化caspase-3及提高原头节内ROS 水平相关,具体机制有待进一步研究。

In vitro effect of glycoglycochenodeoxycholic acid on viability and reactive oxygen species in Echinococcus granulosus protoscoleces

The aim of this study is to investigate the in vitro effect of glycochenodeoxycholic acid (GCDCA) on viability and reactive oxygen species in Echinococcus granulosus protoscoleces. After incubated several days, E.granulosus protoscoleces were randomly allocated into five groups: untreated group, DMSO group, 500 μmol/L group, 1 500 μmol/L group, 2 500 μmol/L group.The effect of GCDCA on protoscoleces viability was investigated using 0.1% eosin staining. Reactive oxygen species (ROS detection kit) was used to detect ROS and Caspase-3 activity was measured using a caspase-3 activity assay kit. Results showed that different concentrations of GCDCA can inhibit protoscoleces' viability, and with prolongation of the time of GCDCA action and increase of its concentration, the inhibitory effect on protoscoleces is gradually increased. In addition, GCDCA increased the mortality of protoscoleces in a concentration-dependent. During the incubation time, the production of ROS and caspase-3 activity clearly increased in protoscoleces treated with GCDCA for 24 hours compared with that in controls (P<0.05). At last, We founded that GCDCA had an inhibitory effect on the growth of E. granulosus protoscoleces, and might be the involvement of caspase-3 activation and ROS production.