羟基喜树碱对细粒棘球蚴原头节的生长抑制作用及其机制

Inhibitory effect of hydroxycamptothecin on protoscoleces of Echinococcus granulosus and its mechanism of action

目的观察羟基喜树碱(HCPT)对体外培养的细粒棘球蚴原头节生存抑制作用及其机制,寻找减少肝脏包虫病术后复发和术中头节外溢的安全高效的可行性药物,并探索其药物作用靶点,为肝脏包虫病的治疗用药提供更多的选择。方法体外培养细粒棘球蚴原头节,分别加入浓度为2.5μmol/L和5μmol/L的HCPT中,37℃恒温孵育2 d和4 d后伊红染色检测原头节生长活力;采用酶标法检测ROS,GSH和caspase-3活性;Western blot检测GRP78和Caspase-12蛋白表达水平。结果 HCPT对细粒棘球蚴生长有抑制作用,HCPT2.5μmol/L和5μmol/L作用4 d,细粒棘球蚴原头节的活力分别为28.2%和7.5%,差异有统计学意义(P<0.05)。高浓度HCPT组ROS和caspase-3含量与对照组比较增高,GSH含量降低(P<0.05);高浓度HCPT作用组GRP78和Caspase-12表达量与对照组比较增加(P<0.05)。结论 HCPT体外可抑制细粒棘球蚴原头节存活,内质网应激机制可能参与其中。

Objective To observe the inhibitory effect of hydroxycamptothecin(HCPT) on protoscoleces of Echinococcus granulosus in vitro and its mechanism of action in order to search for drugs that safely and efficiently reduce the postoperative recurrence of liver hydatid disease and intraoperative protoscolex spillover and to provide more choices for clinical treatment of liver hydatid disease. Methods Protoscoleces of E. granulosus cultured in vitro were used in this study.Protoscoleces were treated with HCPT at a concentration of 2.5, μmol/L or 5 μmol/L and incubated at 37 C for 48 h. Protoscolex activity was detected using eosin staining;ROS, GSH, and caspase-3 activity were detected with reagent kits. GRP-78 and Caspase-12 protein activity was measured using a Western blot assay. Results HCPT inhibited the growth of protoscoleces of E. granulosus, and growth was inhibited at a higher rate with a high concentration of HCPT than with a low concentration. The rate at which growth was inhibition differed significantly(P<0.05). ROS and caspase-3 activity increased significantly while GSH activity decreased with a high concentration of HCPT. Levels of ROS, caspase-3, and GSH activity differed significantly(P<0.05). The level of GRP78 and Caspase-12 expression was significantly higher with a high concentration of HCPT than that with a low concentration. Levels of GRP78 and Caspase-12 expression differed significantly(P<0.05). Conclusion HCPT inhibits the survival of protoscoleces of E. granulosus in vitro, and its mechanism of action may involve endoplasmic reticulum stress.