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目的观察氨基醇咔唑化合物BTB3体外抗细粒棘球蚴的效果。方法体外培养羊源细粒棘球蚴原头节和继发感染小鼠来源的生发层细胞,用浓度为1、2、4、8、10和20μg/ml的BTB3作用3 d后,分别采用美兰染色法和CCK-8法检测原头节和生发层细胞的活性。用扫描电镜观察10μg/ml BTB3对生发层细胞造成的损伤。体外培养细粒棘球蚴囊,用浓度为1、5和10μg/ml的BTB3作用2周后,观察其对囊的影响,并用扫描电镜观察囊内部结构的变化。结果 10μg/ml和20μg/ml BTB3组原头节的死亡率分别为(100.0±0.0)%和(85.2±7.2)%。但当浓度降低后,原头节死亡率均低于10%。生发层细胞经8、10和20μg/ml BTB3作用后,其细胞活性抑制率均达100%,其余低浓度BTB3组的抑制率随浓度的降低而降低。扫描电镜观察发现,BTB3可使生发层细胞发生脱落、皱缩以及空腔化。10μg/ml BTB3作用于棘球蚴囊14 d后,全部囊均出现塌陷。其超微结构显示,BTB3作用后棘球蚴内囊中出现了细胞脱落和不均匀的现象。结论 BTB3对体外培养的细粒棘球蚴原头节、生发层细胞和棘球蚴囊均有较强的作用,是一种潜在的抗棘球蚴药物。
Objective To observe the effect of amino alcohol carbazole compound BTB3 against Echinococcus granulosus in vitro. Methods The germinal layer cells of Echinococcus granulosus and secondary infected mice were cultured in vitro and treated with BTB3 at concentrations of 1, 2, 4, 8, 10 and 20 μg / ml for 3 d respectively. Melanin staining and CCK-8 assay were used to measure the activity of prothallial and germinal cells. Scanning electron microscopy was used to observe the damage to germinal layer cells induced by 10μg / ml BTB3. Echinococcus granulosus was cultured in vitro. The effect of BTB3 at concentrations of 1, 5 and 10 μg / ml for 2 weeks was observed. The changes of the internal structure of the cysts were observed by scanning electron microscopy. Results The mortality rates of the primum nodules in the 10μg / ml and 20μg / ml BTB3 groups were (100.0 ± 0.0)% and (85.2 ± 7.2)%, respectively. However, when the concentration was reduced, the death rate of the primate was less than 10%. The inhibitory rate of germ layer cells treated with BTB3 at concentrations of 8, 10 and 20 μg / ml reached 100%, while the inhibitory rates of other BTB3 low concentration groups decreased with the decrease of concentration. Scanning electron microscopy showed that BTB3 could exfoliate, shrink and cavitate the germinal layer cells. After 10 μg / ml BTB3 was administered to the hydatid cyst for 14 days, all the sacs collapsed. The ultrastructure showed that the cells in the hydatid cyst of BTB3 appeared shedding and non-uniform phenomenon. Conclusion BTB3 is a potential anti-hydatid drug, which has a strong effect on the protoscoleces, germinal layer cells and hydatid cysts of Echinococcus granulosus cultured in vitro.