胆汁对体外培养人细粒棘球蚴原头蚴生长发育的影响

Effect of bile on the growth of human Echinococcus granulosus protoscoleces in vitro

目的探讨胆汁对体外培养人细粒棘球蚴原头蚴生长发育的影响。方法将人细粒棘球蚴原头蚴分别加入纯胆囊胆汁组(Ⅰ组)、RPMI-1640+肝内胆汁(2:1)组(Ⅱ组)及RPMI-1640组(Ⅲ组)中体外孵育。0.1%伊红染色在倒置显微镜下观察原头蚴的形态变化、活动度、顶泡形成情况、头节外翻及成囊发育等生长情况。结果Ⅰ组中95%原头蚴在第9天其外观及内部结构完全破坏;其在胆汁的刺激下第1天活动度达60%,约过半数的头节外翻,随着作用时间的延长,在第7天活动度逐渐降低,原头蚴在第6天后其头节外翻处于停滞状态,大部分原头蚴死亡。Ⅱ组中80%原头蚴在第14天仍能观察到清晰的外观及内部结构;该组原头蚴的活动度在第7天开始下降;头节外翻率随培养时间的增加逐渐增加。Ⅲ组中的原头蚴第14天仍可观察到具有较好的外观及清晰的内部结构;活动度随培养时间的延长而逐渐增加;该组中的原头蚴在第4天开始出现顶泡并随时间的延长出现顶泡的原头蚴数量逐渐增加;头节外翻率逐渐增加。通过连续培养观察,Ⅰ组中的原头蚴无成囊发育;Ⅱ组培养的大部分原头蚴体积缩小,头节内陷,仅少量培养的原头蚴成囊发育,大部分原头蚴随培养时间的延长走向死亡;Ⅲ组中原头蚴逐渐向囊方向发育,且数量逐渐增多。结论 (1)纯胆汁不能在短时间内灭活原头蚴,但随着作用时间的延长原头蚴的活性降低;(2)胆汁可以抑制原头蚴的成囊发育,但在胆汁存在环境下仍有原头蚴成囊。

Objective To explore the effect of bile on the growth of human Echinococcus granulosus protoscoleces in vitro.Methods： The human protoscoleces were cultured in pure bile group（groupⅠ）, RPMI-1640 ＋ bile（2∶1） group（groupⅡ） and RPMI-1640 group（groupⅢ） respectively. 0.1% eosin staining in the inverted microscope to observe the morphological changes of the protoscoleces, activity, the formation of the top bubble, the first section of valgus and cyst development and growth. R esults The results showed that the appearance and internal structure of 95% protoscoleces in groupⅠwere completely destroyed on the 9 th day;The activity of the group was 60% on the first day after bile stimulation, and decreased gradually with the extension effect of time;The group of protoscoleces in the culture process did not observe the formation of the top bubble, and majority of protoscolex were dead. In the groupⅡ, 80% of the protoscoleces can be observed the clear appearance and internal structure on the 14 th day; The activity of the group was decreased on the 7 th day; The number of protoscoleces in evagination was increased gradually with the culture time. In the group Ⅲ, the protoscolices have a good appearance and a clear internal structure on the 14 th day; The activity increased gradually with the prolongation of the culture time. The top bubble of the group began to appear on the 4 th day and the number gradually increased with time. Through the continuous culture observation, the protoscoleces in groupⅠdidn＇t form a hydatid cyst; In groupⅡmost of the protoscoleces shrinked in shape, with a vague internal structure, Only a small number of protoscolices formed cysts, most of protoscolices were in the dormant state; In the GroupⅢ protoscolices gradually toward the direction of the development of the cyst, and the number gradually increased. Conclusion The conclusion is that pure bile could not inactivate the protoscoleces in a short period of time, but the activity decreased with the prolongation of the action time. Bile can inhibit the cystic development of protoscoleces, but protoscoleces can still develop to cyst in the presence of bile.