BMP抑制剂LDN-193189对多房棘球蚴原头节的作用研究

The effect of the BMP inhibitor LDN-193189 on Echinococcus multilocularis protoscoleces

目的研究骨形态发生蛋白(bone morphogenetic protein, BMP)信号通路抑制剂LDN-193189对体外培养多房棘球蚴原头节活性的影响。方法建立体外培养多房棘球蚴体系并随机分为空白对照组、乙醇组、药物组(BMP抑制剂LDN-193189),其中药物组设3个浓度组(1、50、100μmol/L),以不同作用时间(3、6、9和12 d)对原头节进行分组干预试验,用伊红染色和扫描电镜观察各组干预过程中原头节的活性变化,并绘制原头节-药物浓度生存率曲线。结果 LDN-193189干预3、6、9和12 d,各组间存活率差异有统计学意义(P<0.05);100μmol/L LDN-193189组干预3、6 d原头节活性与其余组比较差异有统计学意义(P<0.05);50和100μmol/L LDN-193189组干预9 d时与乙醇组相比原头节活力显著下降(P<0.05);1、50和100μmol/L LDN-193189组干预12 d时原头节活力分别下降(10.65±3.06)%、(36.89±5.15)%和(74.55±6.34)%,50、100μmol/L组与空白对照组(1.23±0.15)%和乙醇组(1.34±0.40)%比较差异均有统计学意义(P<0.01),且原头蚴的死亡率与LDN-193189浓度和干预时间成正比。随着药物浓度的增加,扫描电镜观察原头节顶突变形,小钩脱落,覆盖顶突和体表的微绒毛减少。结论 BMP信号通路抑制剂LDN-193189具有体外抗多房棘球蚴作用,且随浓度的增大和作用时间的延长抑制作用增强。

Objective To investigate the effect of the bone morphogenetic protein signaling pathway inhibitor LDN-193189 on the activity of Echinococcus multilocularis in vitro. Methods A system for the in vitro culture of E. multilocularis was created, and cultured cells were randomly divided into a blank control group, an ethanol group, and a drug group(BMP signaling pathway inhibitor LDN-193189). The drug group was subdivided into three groups: 1, 50, and 100 μmol/L. Protoscoleces were observed at different times(3, 6, 9, and 12 d) at a temperature of 37 ℃ and in an atmosphere of 5% CO2. The activity of and morphological changes in protoscoleces were observed using eosin staining and scanning electron microscopy. Finally, the survival curve for protoscoleces vs. the drug concentration was plotted. Results After intervention with LDN193189 for 3, 6, 9, and 12 d, the survival rate differed significantly among the groups(P<0.05). The activity of protoscoleces in the 100 μmol/L intervention group differed significantly from that in the other groups on days 3 and 6(P<0.05). After 9 d of the intervention, the protoscolex activity in the 50 and 100 μmol/L LDN-193189 groups decreased by 21.40±13.46% and 16.69±1.96%, respectively. Compared to the ethanol group, the protoscolex activity in the 50 and 100 μmol/L LDN-193189 groups decreased significantly(P< 0.05). After 12 d of the intervention, the activity of protoscoleces in the 1, 50, and 100 μmol/L groups decreased(10.65±3.06%, 36.89±5.15%, and 74.55±6.34%). Protoscolex activity in the 50 μmol/L and 100 μmol/L groups differed significantly compared to that in the control group and the ethanol group(P<0.01), and protoscolex mortality was proportional to the concentration of LDN-193189 and the duration of the intervention. Protoscoleces were observed using scanning electron microscopy. As the drug concentration increased, protoscoleces were deformed and small hooks were shed;microvilli covering apical processes and the body surface decreased. Conclusion The BMP signaling pathway inhibitor LDN-193189 has inhibitory action against E. multilocularis in vitro, and this inhibitory action increases with an increase in its concentration and over a longer period of time.