Emergency Department of the First Affiliated Hospital of the Medical College of Shihezi University, Shihezi 832002, China.
Department of Ultrasound Diagnosis, The First Affiliated Hospital, School of Medicine, Shihezi University, Shihezi 832000, China.
Acta Biochim Biophys Sin (Shanghai). 2020 Oct 19;52(10):1140-1147. doi: 10.1093/abbs/gmaa101.
In this study, we investigated the effect of dihydroartemisinin on Echinococcus protoscoleces and explored the role of endoplasmic reticulum stress in this process. Echinococcus protoscoleces were collected and cultured in RPMI 1640 medium. Changes in the expressions of glucose-regulated protein 78 (GRP-78), caspase-12, and C/EBP homologous protein (CHOP) were assessed through confocal immunofluorescence and western blot analysis. Cell viability and morphological changes were observed under a light microscope. The ultrastructure of protoscoleces was observed by scanning electron microscopy and transmission electron microscopy. Caspase-3 activity was detected using an enzyme assay kit. After dihydroartemisinin treatment, the protoscoleces showed loss of viability, and morphological changes including soma contraction, blebs formation, hooks loss, microtrichia destruction, and development of lipid droplets was observed. The levels of caspase-12 and CHOP were increased within 2 days of dihydroartemisinin treatment. However, the levels of GRP-78, caspase-12, and CHOP were decreased in 4 days. Furthermore, caspase-3 activity was increased after treatment with different concentrations of dihydroartemisinin. Dihydroartemisinin can induce apoptosis in protoscoleces via the ER stress-caspase-3 apoptotic pathway in vitro. These results indicate that dihydroartemisinin is a potentially valuable therapeutic agent against echinococcosis.
在这项研究中，我们研究了双氢青蒿素对细粒棘球蚴原头节的影响，并探讨了内质网应激在此过程中的作用。细粒棘球蚴原头节采集并在 RPMI 1640 培养基中培养。通过共聚焦免疫荧光和 Western blot 分析评估葡萄糖调节蛋白 78（GRP-78）、半胱氨酸天冬氨酸蛋白酶-12（caspase-12）和 C/EBP 同源蛋白（CHOP）的表达变化。在光镜下观察细胞活力和形态变化。通过扫描电子显微镜和透射电子显微镜观察原头节的超微结构。使用酶联免疫吸附测定试剂盒检测 caspase-3 活性。双氢青蒿素处理后，原头节活力丧失，并出现形态学变化，包括体收缩、泡形成、钩丢失、微绒毛破坏和脂滴发育。双氢青蒿素处理后 2 天，caspase-12 和 CHOP 水平升高。然而，在 4 天内，GRP-78、caspase-12 和 CHOP 的水平下降。此外，用不同浓度的双氢青蒿素处理后，caspase-3 活性增加。双氢青蒿素可通过内质网应激-caspase-3 凋亡途径诱导原头节体外凋亡。这些结果表明，双氢青蒿素是一种有潜力的抗包虫病治疗药物。
